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Sample records for igg avidity elisa

  1. Use of IgG avidity ELISA to differentiate acute from persistent infection with Salmonella Dublin in cattle

    DEFF Research Database (Denmark)

    Hansen, K.R.; Nielsen, L.R.; Lind, Peter

    2006-01-01

    Aims: To investigate whether an immunoglobulin (Ig)G avidity ELISA can be used to differentiate between acute and persistent infection with Salmonella (S.) Dublin in cattle. To determine whether the IgG isotype, IgG(1) and IgG(2) responses in acute and persistent infections differ. Methods...

  2. IgG avidity to Pseudomonas aeruginosa over the course of chronic lung biofilm infection in cystic fibrosis

    DEFF Research Database (Denmark)

    Mauch, Renan Marrichi; Nørregaard, Lena Lingren; Ciofu, Oana

    2018-01-01

    colonization, using an ELISA technique with the thiocyanate elution method. Avidity was quantitatively determined by calculating the avidity Constant (Kav). Results: IgG avidity to St-Ag significantly increased at the onset (Median Kav = 2.47) and one year after the onset of chronic infection (Median Kav = 3...

  3. IgG avidity test for the diagnosis of acute Toxoplasma gondii infection in early pregnancy.

    Science.gov (United States)

    Pour Abolghasem, Shabnam; Bonyadi, Mohammad Reza; Babaloo, Zohre; Porhasan, Abolfazl; Nagili, Behroz; Gardashkhani, Omid Ali; Salehi, Parviz; Hashemi, Mohammad; Varshoghi, Mojtaba; Gaffari, Gafar Olade

    2011-12-01

    Toxoplasmosis is well known as an important infection in pregnant women. Although many serologic methods are available, diagnosis of early Toxoplasmosis may be extremely difficult. To detect the Toxoplasma IgG antibodies developed at the early stage of infection in pregnant women. 225 pregnant women, who were in the 2nd to 4th month of their pregnancy, enrolled in this study. Anti-toxoplasma IgG, IgM and IgG avidity were evaluated by ELISA method. The patients were categorized into three groups as follows: Group A, 124 cases; IgG+, IgM+, 55.1%; group B, 99 cases; IgG+, IgM-, 44%; and group C, 2 cases; IgG -, IgM +, 0.9%. Fifty five percent of the pregnant women had positive IgG and IgM among which 7.1% had low avidity which revealed an active infection in the pregnant women. In the current study, 44% of pregnant women had positive IgG and negative IgM, all of which had high avidity, which is an indication that in our population the level of toxoplasmosis infection is high and most women have had contacts with this parasite before pregnancy. In this study, the low avidity test was 7.1% showing that the occurrence of toxoplasmosis infection is still a serious issue. Observation of 45.8% high avidity among group A suggests that either IgM has a high half-life or there is a false positive IgM as a result of rheumatologic disorders. Therefore, avidity test is important in predicting maternal toxoplasmosis which is of value in disease treatment.

  4. Evaluation of a Commercial IgE ELISA in Comparison with IgA and IgM ELISAs, IgG Avidity Assay and Complement Fixation for the Diagnosis of Acute Toxoplasmosis

    Czech Academy of Sciences Publication Activity Database

    Kodym, P.; Machala, L.; Roháčová, H.; Širocká, B.; Malý, Marek

    2007-01-01

    Roč. 13, č. 1 (2007), s. 40-47 ISSN 1198-743X Source of funding: V - iné verejné zdroje Keywords : acute infection * diagnosis * IgE ELISA * lymphadenopathy * serology * toxoplasmosis Subject RIV: BB - Applied Statistics, Operational Research Impact factor: 2.980, year: 2007

  5. Clinical usefulness of Western blotting and ELISA avidity for the diagnosis of human toxocariasis.

    Science.gov (United States)

    Rudzińska, M; Kowalewska, B; Sikorska, K

    2017-01-01

    The serodiagnosis of human toxocariasis is difficult. Specific IgGs detected routinely with ELISA based on Toxocara excretory-secretory (TES) antigens often persist for years at an elevated level, which does not allow either the differentiation between an active and persistent infection or monitoring of the effect of treatment. Additionally, false-positive results may occur in co-infections with other helminths due to cross-reactions. We evaluated the usefulness of an IgG avidity index (AI) and a Western blotting (WB) IgG in the diagnosis of patients suspected of Toxocara infection. We studied 138 subjects who were submitted to serological testing two or more times. Confirmation of an infection by WB was achieved in 73.2% of patients. A high AI was obtained in 89.1% of patients, and low AI and borderline AI were found in only 10.9%. Low and borderline values of AI remained at similar levels in subsequent studies over 2-3 years. The results showed the necessity of obligatory verification of all ELISA IgG positive and questionable results by WB. The index of IgG avidity may be helpful in excluding recent infection, but its usefulness in detecting an active phase of invasion requires further research. © 2016 John Wiley & Sons Ltd.

  6. Serological IgG avidity test for ocular toxoplasmosis

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    Suresh S

    2012-01-01

    Full Text Available Subramaniam Suresh1, Saidin Nor-Masniwati1, Muhd Nor Nor-Idahriani1, Wan-Hitam Wan-Hazabbah1, Mohamed Zeehaida2, Embong Zunaina11Department of Ophthalmology, 2Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, MalaysiaBackground: The purpose of this study was to evaluate the immunoglobulin (Ig G avidity of serological toxoplasmosis testing in patients with ocular inflammation and to determine the clinical manifestations of ocular toxoplasmosis.Methods: A retrospective review of all patients presenting with ocular inflammation to the Hospital Universiti Sains Malaysia, Kelantan, Malaysia between 2005 and 2009 was undertaken. Visual acuity, clinical manifestations at presentation, toxoplasmosis antibody testing, and treatment records were analyzed.Results: A total of 130 patients with ocular inflammation were reviewed retrospectively. The patients had a mean age of 38.41 (standard deviation 19.24, range 6–83 years. Seventy-one patients (54.6% were found to be seropositive, of whom five (3.8% were both IgG and IgM positive (suggestive of recently acquired ocular toxoplasmosis while one (0.8% showed IgG avidity ≤40% (suggestive of recently acquired ocular toxoplasmosis and 65 patients (50.0% showed IgG avidity >40% (suggestive of reactivation of toxoplasmosis infection. Chorioretinal scarring as an ocular manifestation was significantly more common in patients with seropositive toxoplasmosis (P = 0.036. Eighteen patients (13.8% were diagnosed as having recent and/or active ocular toxoplasmosis based on clinical manifestations and serological testing.Conclusion: Ocular toxoplasmosis is a clinical diagnosis, but specific toxoplasmosis antibody testing helps to support the diagnosis and to differentiate between reactivation of infection and recently acquired ocular toxoplasmosis.Keywords: ocular toxoplasmosis, chorioretinal scar, toxoplasmosis antibody, IgG avidity test

  7. Indeterminate/moderate IgG avidity during HCMV infection: comparison of methods

    Directory of Open Access Journals (Sweden)

    Massimo De Paschale

    2010-03-01

    Full Text Available Background. The IgG avidity test is usually used for differentiating between primary and non-primary HCMV infection within 3 months. Weak avidity is highly suggestive of a primary infection, while high avidity tends to exclude it.An indeterminate or moderate avidity, however, does not allow a clear dating. Since there are several avidity tests with different performances, those tests that are able to minimize the results with indeterminate/moderate avidity are particularly useful. Objectives. The aim of our work was to evaluate the results obtained with two IgG avidity tests in IgG and IgM anti-HCMV positive patients. Study Design. 113 anti-HCMV IgG and IgM positive samples were tested with Enzime Linked Fluorescent Assay (ELFA and Chemiluminescence Immuno Assay (CLIA IgG avidity test. Results. 21 samples (18.6%, 50 (44.2% and 42 (37.2% with ELFA and 53 samples (46.9%, 10 (8.8% and 50 (44.2% with CLIA were found to have respectively low, indeterminate/moderate and high avidity. Of the 50 ELFA indeterminate avidity samples, 32 (64%, 10 (20% and 8 (16% were found to have respectively low, moderate and high CLIA.avidity. For 11 cases of the 32 ELFA indeterminate avidity and CLIA low avidity, there were previous data showing a seroconversion within three months. In a case of the 8 ELFA indeterminate avidity and CLIA high avidity, there were, instead, previous data of IgG and IgM positivity already four months earlier. Conclusions. It appears that the CLIA test for IgG avidity is more effective than ELFA for dating HCMV infection.

  8. Diagnosis of acute toxoplasmosis in pregnant women referred to therapeutic centers of Alborz Province (Iran using immunoglobulin G avidity ELISA technique

    Directory of Open Access Journals (Sweden)

    Lame Akhlaghi

    2016-11-01

    Full Text Available Objective: To evaluate immunoglobulin G (IgG avidity as a useful and reliable technique in diagnosing toxoplasmosis in pregnant women referring to therapeutic centers of Alborz Province (Iran in 2014, against two other tests, IgG and immunoglobulin G (IgM antiToxoplasma. Methods: Serum samples (468 in total were obtained from different therapeutic centers in Karaj City. ELISA method was used to test the anti-Toxoplasma avidity of IgG, IgM and IgG. The data were analyzed by descriptive statistical methods and Chi-square test (P < 0.05 using SPSS 17.0. Results: Anti-Toxoplasma tests of IgM and IgG were positive in 9 and 86 samples respectively. Also, a borderline IgM was detected in 2 suspected samples. In addition, among all positive and suspected samples, 79 cases indicated high titers of IgG avidity, 7 cases were of low titers and 1 case was of a borderline titer. The prevalence of anti-Toxoplasma antibodies was 20%. The sera which showed high avidity index was obtained from patients at chronic phase of infection (92% while those which showed low avidity levels were from patients at acute toxoplasmosis (77.7%. Conclusions: This study clearly showed that acute and chronic phases of toxoplasmosis could be differentiated with the aid of IgG avidity test. This test may also assist in recognizing old and newly acquired infections.

  9. Contribution of IgG avidity and PCR for the early diagnosis of ...

    African Journals Online (AJOL)

    Background: Acute toxoplasmosis in pregnant women presents a high risk of Toxoplasma transmission to the fetus. Early diagnosis is difficult, especially when serological testing for IgG/IgM antibodies fail to differentiate between a recent and a past infection. In this case, we rely on IgG avidity or PCR assays. Objectives: The ...

  10. Contribution of IgG avidity and PCR for the early diagnosis of ...

    African Journals Online (AJOL)

    2017-09-03

    Sep 3, 2017 ... Abstract: Background: Acute toxoplasmosis in pregnant women presents a high risk of Toxoplasma transmission to the fetus. Early di- agnosis is difficult, especially when serological testing for IgG/IgM antibodies fail to differentiate between a recent and a past infection. In this case, we rely on IgG avidity or ...

  11. Generation of polyclonal antibody with high avidity to rosuvastatin and its use in development of highly sensitive ELISA for determination of rosuvastatin in plasma

    Directory of Open Access Journals (Sweden)

    Al-Malaq Hamoud A

    2011-07-01

    Full Text Available Abstract In this study, a polyclonal antibody with high avidity and specificity to the potent hypocholesterolaemic agent rosuvastatin (ROS has been prepared and used in the development of highly sensitive enzyme-linked immunosorbent assay (ELISA for determination of ROS in plasma. ROS was coupled to keyhole limpt hemocyanin (KLH and bovine serum albumin (BSA using carbodiimide reagent. ROS-KLH conjugate was used for immunization of female 8-weeks old New Zealand white rabbits. The immune response of the rabbits was monitored by direct ELISA using ROS-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and avidity to ROS was scarified and its sera were collected. The IgG fraction was isolated and purified by avidity chromatography on protein A column. The purified antibody showed high avidity to ROS; IC50 = 0.4 ng/ml. The specificity of the antibody for ROS was evaluated by indirect ELISA using various competitors from the ROS-structural analogues and the therapeutic agents used with ROS in a combination therapy. The proposed ELISA involved a competitive binding reaction between ROS, in plasma sample, and the immobilized ROS-BSA for the binding sites on a limited amount of the anti-ROS antibody. The bound anti-ROS antibody was quantified with horseradish peroxidase-labeled second anti-rabbit IgG antibody (HRP-IgG and 3,3',5,5'-tetramethylbenzidine (TMB as a substrate for the peroxidase enzyme. The concentration of ROS in the sample was quantified by its ability to inhibit the binding of the anti-ROS antibody to the immobilized ROS-BSA and subsequently the color intensity in the assay wells. The assay enabled the determination of ROS in plasma at concentrations as low as 40 pg/ml.

  12. Persistent Low Toxoplasma IgG Avidity Is Common in Pregnancy: Experience from Antenatal Testing in Norway.

    Science.gov (United States)

    Findal, Gry; Stray-Pedersen, Babill; Holter, Ellen K; Berge, Tone; Jenum, Pål A

    2015-01-01

    The parasite Toxoplasma gondii might harm the fetus if a woman is infected during pregnancy. IgG seroconversion and significant increase in IgG antibody amount in pregnancy indicates maternal infection. Presence of toxoplasma immunoglobulin M (IgM), immunoglobulin G (IgG) and low IgG avidity in a single serum sample indicates possible maternal infection, but positive toxoplasma IgM and low IgG avidity may persist for months and even years. We aimed to evaluate avidity development during pregnancy in a retrospective study. Serial blood samples from 176 pregnant women admitted to Oslo University Hospital 1993-2013 for amniocentesis because of suspected toxoplasma infection were included. Data were obtained from journals and laboratory records. The avidity method used was based on Platelia Toxo IgG assay. Mean maternal age at first serology was 29.9 years (SD 5.2, range 18-42). In 37 (21%) women only the avidity increased from low to high in toxoplasma serology should ideally be collected in early pregnancy and if stable values of toxoplasma IgM and low IgG-avidity are detected in a second sample after three to four weeks, the need for amniocentesis can be questioned.

  13. Performance Evaluation of the VIDAS® Measles IgG Assay and Its Diagnostic Value for Measuring IgG Antibody Avidity in Measles Virus Infection

    Science.gov (United States)

    Dina, Julia; Creveuil, Christian; Gouarin, Stephanie; Viron, Florent; Hebert, Amelie; Freymuth, Francois; Vabret, Astrid

    2016-01-01

    The objective of this study is primarily to compare the performance of the VIDAS® Measles immunoglobulin (Ig)G assay to that of two other serological assays using an immunoassay technique, Enzygnost® Anti-measles Virus/IgG (Siemens) and Measles IgG CAPTURE EIA® (Microimmune). The sensitivity and the agreement of the VIDAS® Measles IgG assay compared to the Enzygnost® Anti-measles Virus/IgG assay and the Measles IgG CAPTURE EIA® assay are 100%, 97.2% and 99.0%, 98.4%, respectively. The very low number of negative sera for IgG antibodies does not allow calculation of specificity. As a secondary objective, we have evaluated the ability of the VIDAS® Measles IgG assay to measure anti-measles virus IgG antibody avidity with the help of the VIDAS® CMV IgG Avidity reagent, using 76 sera from subjects with measles and 238 other sera. Different groups of populations were analyzed. In the primary infection measles group, the mean IgG avidity index was 0.16 (range of 0.07 to 0.93) compared to 0.79 (range of 0.25 to 1) in the serum group positive for IgG antibodies and negative for IgM. These data allow to define a weak anti-measles virus IgG antibody avidity as an avidity index (AI) 0.6. The VIDAS® Measles IgG assay has a performance equivalent to that of other available products. Its use, individual and quick, is well adapted to testing for anti-measles immunity in exposed subjects. PMID:27556477

  14. Serodiagnosis of Toxoplasmosis: The effect of measurement of IgG avidity in pregnant women in Rabat in Morocco.

    Science.gov (United States)

    Laboudi, Majda; Sadak, Abderrahim

    2017-08-01

    The diagnosis of Toxoplasmosis in pregnant women during the early first trimester of pregnancy is very important for preventing congenital infection of the fetus; it will not only prevent the risk of transmitting the infection to the fetus but it will also enable to give these women a preventive treatment. In this study, the avidity test was performed on pregnant women during their first prenatal visit at the National Institute of Hygiene in Rabat, Morocco. One hundred and twenty-eight sera samples were collected from 128 pregnant women between August 2015 and June 2016; these women were chosen retrospectively and were in their first four months of pregnancy. The samples were screened using the specific anti-Toxoplasma IgG and IgM antibodies and were subjected to an IgG avidity test. After the serological screening, only 54 women (42.4%) were tested positive for IgG antibodies and five women (3.9%) were tested positive for both anti-Toxoplasma IgG and IgM antibodies. Four IgM-negative women had low-avidity antibodies. However, none of the IgG-avidity test had detected low-avidity antibodies in the five IgM-positive women; three women (60%) had high-avidity antibodies, indicating that the infection was acquired in the distant past. The avidity test is a helpful tool to exclude a recently acquired toxoplasmosis infection within IgM-positive serum samples in pregnant women during their first trimester of pregnancy. Thus, allowing to perform an appropriate therapeutic intervention. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. A new human IgG avidity test, using mixtures of recombinant antigens (rROP1, rSAG2, rGRA6), for the diagnosis of difficult-to-identify phases of toxoplasmosis.

    Science.gov (United States)

    Drapała, Dorota; Holec-Gąsior, Lucyna; Kur, Józef; Ferra, Bartłomiej; Hiszczyńska-Sawicka, Elżbieta; Lautenbach, Dariusz

    2014-07-01

    The preliminary diagnostic utility of two mixtures of Toxoplasma gondii recombinant antigens (rROP1+rSAG2 and rROP1+rGRA6) in IgG ELISA and IgG avidity test has been evaluated. A total of 173 serum samples from patients with toxoplasmosis and seronegative people were examined. The sensitivity of IgG ELISA for rROP1+rSAG2 and rROP1+rGRA6 was 91.1% and 76.7%, respectively, while the reactivity for sera from patients where acute toxoplasmosis was suspected was higher, at 100% and 95.4%, respectively, than for people with chronic infection, at 88.2% and 70.6%. In this study a different trend in avidity maturation of IgG antibodies for two mixtures of proteins in comparison with native antigen was observed. The results suggest that a new IgG avidity test using the mixtures of recombinant antigens may be useful for the diagnosis of difficult-to-identify phases of toxoplasmosis. For this reason, selected mixtures after the additional tests on groups of sera with well-defined dates of infection could be used as a better alternative to the native antigens of the parasite in the serodiagnosis of human T. gondii infection. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Changes in avidity and specificity of IgG during electro-oxidation. Relevance of binding of antibodies to beta2-GPI.

    Science.gov (United States)

    Bozic, B; Cucnik, S; Kveder, T; Rozman, B

    2006-11-01

    The immune response may be changed due to altered proteins or modifications of immunoglobulins, including oxidative processes. The susceptibility to oxidative modifications depends greatly on amino-acid moiety composition due to chemical characteristics (instability) of their side-chains. Initial steps of oxidation may change the specificity and avidity of immunoglobulins due to chemical alteration of the hypervariable region. The oxidation of antibodies increases the hydrophilic nature of the paratopes and makes them more susceptible for the binding to cationic surfaces even without the strong surface-to-surface fitting. The electro-oxidation of IgG significantly changes the immunoreactivity and specificity of IgG fractions, regardless of the initial immunoreactivity to a specific autoantigen also in healthy persons. Data are presented on changes in the immunoreactivity as well as the avidity of antibodies against beta2-glycoprotein I after being exposed to direct current. ELISA measurements showed increased reactivity of anti-beta2-glycoprotein I antibodies at the beginning and various, fluctuating results after prolonged exposure to electro-oxidation. Inter-individual differences in chemical stability of immunoglobulins and patient's antioxidative status may influence the range of their alterations and their impact on health/disease balance.

  17. High Concentrations of Measles Neutralizing Antibodies and High-Avidity Measles IgG Accurately Identify Measles Reinfection Cases

    Science.gov (United States)

    Rota, Jennifer S.; Hickman, Carole J.; Mercader, Sara; Redd, Susan; McNall, Rebecca J.; Williams, Nobia; McGrew, Marcia; Walls, M. Laura; Rota, Paul A.; Bellini, William J.

    2016-01-01

    In the United States, approximately 9% of the measles cases reported from 2012 to 2014 occurred in vaccinated individuals. Laboratory confirmation of measles in vaccinated individuals is challenging since IgM assays can give inconclusive results. Although a positive reverse transcription (RT)-PCR assay result from an appropriately timed specimen can provide confirmation, negative results may not rule out a highly suspicious case. Detection of high-avidity measles IgG in serum samples provides laboratory evidence of a past immunologic response to measles from natural infection or immunization. High concentrations of measles neutralizing antibody have been observed by plaque reduction neutralization (PRN) assays among confirmed measles cases with high-avidity IgG, referred to here as reinfection cases (RICs). In this study, we evaluated the utility of measuring levels of measles neutralizing antibody to distinguish RICs from noncases by receiver operating characteristic curve analysis. Single and paired serum samples with high-avidity measles IgG from suspected measles cases submitted to the CDC for routine surveillance were used for the analysis. The RICs were confirmed by a 4-fold rise in PRN titer or by RT-quantitative PCR (RT-qPCR) assay, while the noncases were negative by both assays. Discrimination accuracy was high with serum samples collected ≥3 days after rash onset (area under the curve, 0.953; 95% confidence interval [CI], 0.854 to 0.993). Measles neutralizing antibody concentrations of ≥40,000 mIU/ml identified RICs with 90% sensitivity (95% CI, 74 to 98%) and 100% specificity (95% CI, 82 to 100%). Therefore, when serological or RT-qPCR results are unavailable or inconclusive, suspected measles cases with high-avidity measles IgG can be confirmed as RICs by measles neutralizing antibody concentrations of ≥40,000 mIU/ml. PMID:27335386

  18. The evaluation of rubella and sitomegalovirus IgG avidity tests in pregnants: four-year experience

    Directory of Open Access Journals (Sweden)

    Berrin Uzun

    2014-09-01

    Full Text Available Objective: In this study, we aim to evaluate the retrospective IgG avidity index (AI test results of the pregnant, who have applied to our hospital and had pregnancy screening test and have been asked for rubella and cytomegalovirus antibody tests and IgG AI test in 4 year period. Methods: Anti-rubella IgM, anti-rubella IgG, anti-CMV IgM and anti-CMV antibodies were performed by IMMULITE 2000XPi™ Immunoassay System (Siemens, Germany whereas IgG AI tests were performed by commercial kits of rubella and cytomegalovirus antibody tests (Dia.Pro® Diagnostic, Milano-Italy. Results: Between January 2010 and December 2013, 23 (7.32 % of 314 pregnant women tested rubella IgG AI had low AI, and 266 cases had high AI rate; 14 (17.28% of 81 pregnant women tested CMV IgG AI, had lower AI rate, 52 of them had high AI. 9 out of 23 pregnant women, who were detected low rubella were requested only AI without any IgM and IgG value; the IgG antibody of 13 pregnant was reactive, and IgM antibody was not requested; IgG and IgM antibody of 1 pregnant was detected positive. When 5 pregnant, who were detected low CMV AI were requested AI without IgG and IgM value; the IgG and IgM of 4 was reactive; 4 of them had IgG reactive but IgM negative; 1 had IgG and IgM negative. Conclusion: As a result, our low rubella AI rate has been detected lower than the rates in our country since our study population contains only the pregnant. Our low CMV AI rate varies due to the same reasons as literature data. During pregnancy scanning, performing of AI testing together with specific IgG and IgM will save both time and will be more meaningful in clinical evaluation for pregnant women. J Clin Exp Invest 2014; 5 (3: 420-423

  19. Performance of the multitarget Mikrogen Chlamydia trachomatis IgG ELISA in the prediction of tubal factor infertility (TFI) in subfertile women : Comparison with the Medac MOMP IgG ELISA plus

    NARCIS (Netherlands)

    van Ess, Eleanne F.; Ouburg, Sander; Spaargaren, Joke; Land, Jolande A.; Morre, Servaas A.

    2017-01-01

    There is a need for more accurate Chlamydia trachomatis (CT) IgG antibody tests for tubal factor infertility (TFI) diagnostics. We evaluated the predictive value for TFI of Medac ELISA plus (MOMP) and multitarget Mikrogen ELISA (MOMP-CPAF-TARP). Based on Medac ELISA plus results, 183 subfertile

  20. DETECTION OF HUMAN ANTI-ZIKA VIRUS IgG BY ELISA USING AN ANTIGEN FROM in vitro INFECTED VERO CELLS: PRELIMINARY RESULTS

    Directory of Open Access Journals (Sweden)

    Laura Masami SUMITA

    Full Text Available SUMMARY Zika virus (ZKV infection is a huge public health problem in Brazil because of the increased incidence of microcephaly in neonates from infected mothers. Detection of specific IgG antibodies in maternal serum samples constitutes an important approach for diagnosing ZKV infection and evaluating its relationship with neonatal microcephaly. However, as there is no serological test produced in Brazil to detect IgM and IgG antibodies against ZKV, we sought to examine specific IgG in serum samples from patients or suspected mothers to detect previous infection and to test for specificity with regard to flaviviral infections occurring in the same area. Brazilian Zika virus native antigens were obtained from infected Vero cell layers or free virions in the culture medium and then used in ELISA. We tested sera from eight ZKV RNA-diagnosed infected patients (ZKVR, seven neonates with microcephaly and their mothers after delivery (MM, 140 dengue virus IgM-positive (DM and IgG (DG-positive patients, and 100 yellow fever (YF-vaccinated patients. According to the ELISA, ZKVR samples were mostly positive (7/8, and all the MM serum samples were positive for ZKV IgG (7/7. In contrast, cross-reactions for dengue or yellow fever-vaccinated patients were observed, including DM (48/95, DG (10/45 or YF (3/100 serum samples; however, these cross-reactions exhibited low antigen avidity so that 6 M urea largely removed this cross-reactivity, with only a few cross-reacting samples remaining (8/140. ELISA based on extracted virions was much more specific, with all ZKVR (8/8 and MM sera being positive for ZKV IgG (7/7 and only borderline cross-reactivity found for DM (6/95, DG (3/45 or YF (4/100-vaccinated serum samples. This technique (ELISA can identify specific IgG in ZKV-infected patients and may be helpful in diagnosing congenital infetions after maternal RNA virus clearance or in epidemiological studies.

  1. [Effect comparison between two ELISA kits in IgG antibody detection of Echinococcus granulosus].

    Science.gov (United States)

    Chu, Yan-Hong; Cai, Yu-Chun; Ai, Lin; Lu, Yan; Zhang, Jia; Chen, Jia-Xu

    2013-06-01

    To compare the effects of two ELISA kits on IgG antibody detection of human Echinococcus granulosus. A Total of 134 sera of patients with echinococcosis, paragonimiasis westermani, clonorchiasis sinensis, schistosomiasis japonica, and cysticercosis cellulosae, and normal persons were detected by two IgG ELISA kits produced by different companies. Furthermore, the specificity, sensitivity and cross reactivity were counted and analyzed statistically. The sensitivity and specificity were extremely high of the two kits as 100.00%. The cross-reactivity rates were 25.00% (paragonimiasis westermani), 26.09% (clonorchiasis sinensis), 10.00% (schistosomiasis japonica), and 87.5% (cysticercosis), respectively, by using the kit produced by the Combined Company in Shenzhen; the cross-reactivity rates were 5.00% (paragonimiasis westermani), 13.04% (clonorchiasis sinensis), 20.00% (schistosomiasis japonica), and 93.75% (cysticercosis) respectively, by using the kit produced by Haitai Company in Zhuhai. In addition, there was a significant difference of Paragonimus westermani detection (P 0.05) between the two kits. Both ELISA kits on IgG antibody detection of human Echinococcus granulosus have the advantages of a high sensitivity, specificity, convenience and high-speed. However, it is also in urgent need to further solve the cross-reactivity of Echinococcus granulosus with other parasites, in order to improve the accuracy of early diagnosis.

  2. Indirect ELISA for the detection of IgG specific to Newcastle disease virus in quail serum

    OpenAIRE

    Oliveira, D.D.; Folgueras-Flatschart, A.V.; Flatschart, R.B.; Resende, J.S.; Abreu, J.T.; Martins, N.R.S.

    2007-01-01

    An indirect ELISA for the detection of japanese quail IgG specific to Newcastle disease virus (NDV) was developed. The secondary anti-quail IgG was produced in Balb/c mice, by inoculating Freund's complete adjuvant emulsified japanese quail-IgG extract. The purification of IgG was achieved using the caprilic acid method. The ELISA was compared to the haemagglutination-inhibition (HI) test for antibodies to NDV. ELISA cut-off point was established through TG-ROC analysis. Total correlation was...

  3. Assessment of the diagnostic performance of the IDS-iSYS tests for toxo IgG, toxo IgM and avidity.

    Science.gov (United States)

    Levigne, Pauline; Peyron, François; Wallon, Martine

    2016-10-01

    When acquired during pregnancy toxoplasmosis can have devastating consequences on the fetus. As maternal infection is in the majority of cases subclinical, the diagnosis of toxoplasmosis relies on serological tests for the detection of IgG, IGM and the mesure of IgG avidity. We evaluated the performance of IDS-iSYS a new automatized instrument based on chemiluminescence for the diagnosis of the disease. Our study was based on non-selected samples received in our laboratory either for the determination of serological status or for distinguishing acute from chronic infection. Seven hundred eighty three samples were enrolled in the study. Compared with Architect IgG and IgM assays, the sensitivity and specificity were respectively 99% and 99% for IgG, and 75% and 97% for IgM. We observed higher remaining titers for IDS iSYS IgG, which could reduce the proportions of patients who have to be retested because of doubtful titers. IgM detection and avidity scored equivalent performance with both methods. This automate appears to be a reliable and easy-to-use tool for diagnosing toxoplasmosis in different clinical settings. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

    OpenAIRE

    Villa-Mancera, Abel; Molina-Mendoza, Pedro; Hern?ndez-Guzm?n, Karina; Olivares-P?rez, Jaime; Sarracent-P?rez, Jorge; Zumaquero-R?os, Jos?

    2016-01-01

    The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELIS...

  5. Comparison of in-house IgM and IgG ELISAs for the serodiagnosis of melioidosis in Malaysia.

    Science.gov (United States)

    Hii, Shirley Yi Fen; Ali, Noor Azila; Ahmad, Norazah; Amran, Fairuz

    2017-11-01

    Melioidosis is an endemic infectious disease in Southeast Asia and northern Australia, caused by Burkholderia pseudomallei. However, the incidence rate in Malaysia is not well documented. The high mortality rate and broad range of clinical presentations require rapid and accurate diagnosis for appropriate treatment. This study compared the efficacy of in-house IgM and IgG ELISA methods using a local B. pseudomallei strain. The diagnostic accuracy of the in-house IgG ELISA was better than that of the IgM ELISA: sensitivity (IgG: 84.71 %, IgM: 76.14 %) and specificity (IgG: 93.64 %, IgM: 90.17 %); positive predictive value (IgG: 86.75 %, IgM: 79.76 %) and negative predictive value (IgG: 92.57 %, IgM: 89.66 %); likelihood ratio (LR) [IgG: 13.32, IgM: 7.75 (LR+); IgG: 0.16, IgM: 0.26 (LR-)], and was supported by the observation of the absorbance value in comparisons between culture and serology sampling. In-house IgG ELISA was shown to be useful as an early diagnostic tool for melioidosis.

  6. Development and application of an ELISA for the detection of porcine deltacoronavirus IgG antibodies.

    Directory of Open Access Journals (Sweden)

    Anil Thachil

    Full Text Available Porcine deltacoronavirus (PDCoV, also known as porcine coronavirus HKU15, was first detected in North America in early 2014 and associated with enteric disease in pigs, resulting in an urgent need to further investigate the ecology of this virus. While assays detecting nucleic acids were implemented quickly, assays to detect anti-PDCoV antibodies have not been available. In this study, an indirect anti-PDCoV IgG enzyme-linked immunosorbent assay (ELISA based on the putative S1 portion of the spike protein was developed and utilized to determine the prevalence of anti-PDCoV IgG in U.S. pigs. The diagnostic sensitivity of the PDCoV ELISA was 91% with a diagnostic specificity of 95%. A total of 968 serum samples were tested including samples with confirmed infection with PDCoV, porcine epidemic diarrhea virus (PEDV, transmissible gastroenteritis virus or porcine respiratory coronavirus. There was no cross-reactivity with any of the other coronaviruses. Among 355 arbitrarily selected serum samples collected in 2014 and originating from 51 farms across 18 U.S. states, anti-PDCoV IgG antibodies were detected in 8.7% of the samples and in 25.5% of the farms whereas anti-PEDV IgG was detected in 22.8% of the samples and in 54.9% of the farms. In addition, anti-PDCoV IgG antibodies were detected in archived samples collected in 2010, perhaps indicating an earlier undetected introduction into the U.S. pig population. Overall, the obtained data suggest that PDCoV seroprevalence in U.S. pigs is lower compared to PEDV and PDCoV may have been introduced to the U.S. prior to PEDV.

  7. Significance of IgG optical density ratios (index value) in single reactive anti-Dengue virus IgG capture ELISA.

    Science.gov (United States)

    Chatterjee, Shiv Sekhar; Sharma, Ankush; Choudhury, Shilpee; Chumber, Sushil Kumar; Kaur, Mandeep; Bage, Ras; Parkhe, Nittin; Khanduri, Uma

    2016-12-01

    A single reactive IgG anti-Dengue virus ELISA test in the absence of IgM antibodies or NS1 antigen may denote current infection or past exposure to the virus. To determine whether IgG index value can be used to identify true current dengue infection we conducted a prospective observational study. Suspected dengue patients (n =1745) were tested in their first specimen by MAC-ELISA, GAC-ELISA and NS1 antigen ELISA. Patients with MAC-ELISA and NS1Antigen non-reactive but GAC-ELISA reactive results (n =57) in their first test were followed up and repeated sampling was asked for IgG index values were calculated according to the manufacturer's instruction and classified as: low (2.2-2.5), medium (2.5-4.0) and high (>4.0). 16 out of 57 patients (28.1%) had low IgG Index value whereas 26 cases (45.6%) were categorized as medium and 15(26.3%) were classified as patients with high IgG index. Nine patients with paired reactive serology or antigen positive status were categorised as serologically confirmed dengue fever, 11 patients as not dengue with categorical evidence of other infections while the rest 37 casas with clinical, radiological and laboratory parameters suggestive of dengue but no serological confirmation as possible dengue. Among confirmed, possible and non-Dengue cases, 33.3, 32.4 and 0.0% had high Index value in comparison with 22.2, 29.7 and 27.3% showing low Index values, respectively. Our results suggested a high IgG response in favour of true dengue infection than past exposure while no conclusions should drawn from a low or medium reactive GAC-ELISA results in the absence of IgM antibodies and NS1 Ag.

  8. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

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    Abel Villa-Mancera

    2016-01-01

    Full Text Available The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P<0.01 for low seropositivity (r=0.93 and medium seropositivity (r=0.84. The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans.

  9. Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

    Science.gov (United States)

    Molina-Mendoza, Pedro; Hernández-Guzmán, Karina; Olivares-Pérez, Jaime; Sarracent-Pérez, Jorge; Zumaquero-Ríos, José

    2016-01-01

    The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P < 0.01) for low seropositivity (r = 0.93) and medium seropositivity (r = 0.84). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans. PMID:27563665

  10. Seroprevalencia y detección de infección primaria por citomegalovirus mediante prueba de avidez IgG en el primer trimestre de embarazo

    Directory of Open Access Journals (Sweden)

    Conne L González-García

    2014-11-01

    Full Text Available Objective. To determine the seroprevalence and detection of primary infection by cytomegalovirus (CMV with immunoglobulin G (IgG avidity test during the first quarter of pregnancy in the General Hospital in Morelia, Michoacan. Materials and methods. A total of 177 patients were studied employing a modified Elisa test using a chemiluminescent microparticle immunoassay (CMIA for the detection of CMV antibodies (IgG and immunoglobulin M [IgM], and IgG avidity. Results. 90.4% were positive for IgG, and of these, 2.3% were also reactive for IgM, and in this group the IgG avidity test reported low avidity for 1.1% and higher avidity in the same percentage. 9.6% were seronegative. Conclusions. Similarity was found with published studies in Mexico. Health professionals should know the clinical algorithms for diagnosis and proper management of CMV infection using the IgG avidity test.

  11. [The comparison of two newborn cytomegalovirus IgG antibody screening ELISA kits].

    Science.gov (United States)

    Zhang, Shun-Xian; He, Xiao-Zhou; Wang, Shi-Wen; Wang, Xiao-Fang

    2013-10-01

    This study compared two newborn Cytomegalovirus (CMV) IgG antibody screening ELISA kits and evaluated the detection effectiveness of Abnova kit. CMV IgG antibodies were detected by both SeraQuest and Abnova kits from dried blood spot (DBS) samples of 488 newborn heel sticks. The detection abilities of these two kits were compared in different sample dilution concentrations. Relative detection effectiveness of the Abnova kit was defined by statistical method using the SeraQuest kit as a point of comparison. Compared to the SeraQuest screening test kit, the Abnova kit revealed a sensitivity of 98.9%, specificity of 78.6%, positive predictive value of 99.3%, negative predictive value of 68.8%, and the coincidence rate for these two screening test kits at 98.3%. The consistency check of both kits based on interpretation of the kappa statistic was relatively good. For the Abnova kit, the "area under the ROC curve" was 0.887, which indicates moderate accuracy. Abnova kit can be applied to newborn screening for congenital CMV infections. However, repeating the test for ambiguous results is suggested to increase the specificity and negative predictive value.

  12. ELISA indireto para detecção de IgG antivírus da doença de Newcastle em soro de codorna Indirect ELISA for the detection of IgG specific to Newcastle disease virus in quail serum

    Directory of Open Access Journals (Sweden)

    D.D. Oliveira

    2007-10-01

    Full Text Available An indirect ELISA for the detection of japanese quail IgG specific to Newcastle disease virus (NDV was developed. The secondary anti-quail IgG was produced in Balb/c mice, by inoculating Freund's complete adjuvant emulsified japanese quail-IgG extract. The purification of IgG was achieved using the caprilic acid method. The ELISA was compared to the haemagglutination-inhibition (HI test for antibodies to NDV. ELISA cut-off point was established through TG-ROC analysis. Total correlation was observed between the ELISA and the HI, being the ELISA efficient in the identification of positive and negative sera, with high sensitivity and specificity (100%. These results validate the use of the indirect ELISA as an alternative for the detection of NDV-specific IgG in japanese quail sera, with the advantage of high sensitivity and automation.

  13. Caracterización de IgM, IgG total, IgG1 y anticuerpos de cadena pesada en calostro de llamas (Lama Glama) mediante Elisa

    OpenAIRE

    Caggiano, Nicolás; Saccodossi, Natalia; Gentile, Maria Teresa; Chiappe Barbará, María Angelina; Leoni, Juliana; de Simone, Emilio Adrian

    2016-01-01

    Objetivos: determinar los niveles de IgM, IgG total y Anticuerpos de Cadena Pesada (HCAbs; por su sigla en inglés Heavy Chain Antibodies) (IgG2 e IgG3) en calostro de llamas y evaluar la concentración de HCAbs en relación a la IgG total y al isotipo convencional IgG1. Métodos: en este estudio se utilizaron 15 llamas preñadas, que fueron ordeñadas dentro de las primeras 24 horas post-parto. Se diseñaron ELISAs Sandwich para la cuantificación de IgM total, IgG total e IgG1. La concent...

  14. IgG binding of mugwort pollen allergens and allergoids exposed to simulated gastrointestinal conditions measured by a self-developed ELISA test

    Directory of Open Access Journals (Sweden)

    RATKO M. JANKOV

    2004-07-01

    Full Text Available This study considers the influence of exposure to simulated gastrointestinal conditions (saliva, gut, intestine and acidic conditions of the gut on IgG binding of unmodified allergens and three types of LMW allergoids of Artemisia vulgaris pollen extract obtained by means of potassium cyanate, succinic and maleic anhydride. It also concerns the optimization of a self-developed ELISA assay for comparison of the specific IgG binding of mugwort pollen extract and modified mugwort pollen derivatives. The ELISA was conducted with a mugwort pollen extract coupled to the plate, using the sera from 12 mugwort-pollen allergic patients. The exposure to saliva fluid for 2 min did not influence the IgG binding properties of allergens and allergoids. Exposure of mugwort pollen allergens and LMW allergoids to the acidic conditions of the gut did not dramatically change their IgG binding properties. By exposing mugwort pollen extract and LMW derivatives to the SGF conditions for 1 h, the percent of IgG binding epitopes was reduced to a half of its starting value in the extract and to about 30 % in all the allergoid samples. After prolonged exposure only the carbamyl derivative showed reduced IgG binding. Changes of the IgG binding potential of all four samples after exposure in SIF followed a similar pattern.

  15. Validation and evaluation of VapA-specific IgG and IgG subclass enzyme-linked immunosorbent assays (ELISAs) to identify foals with Rhodococcus equi pneumonia.

    Science.gov (United States)

    Sanz, M G; Oliveira, A F; Loynachan, A; Page, A; Svansson, V; Giguère, S; Horohov, D W

    2016-01-01

    Rhodococcus equi (Rhodococcus hoagii/Prescottella equi) is a common cause of foal pneumonia, but its diagnosis remains a challenge for equine veterinarians. While the VapA-specific (virulence-associated protein A) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) has low sensitivity and specificity for detecting pneumonic foals, little is known about VapA-specific IgG subclasses. To evaluate the performance of VapA-specific ELISA for IgG and its subclasses IgGa, IgGb and IgG(T) in the early diagnosis of pneumonia caused by R. equi. Assay validation followed by assessment of diagnostic performance using archived samples from animals of known status. Serum samples from exposed (n = 125) and nonexposed adult horses (n = 10) and from experimentally challenged and naturally infected foals were used for ELISA validation. Post mortem and tissue culture records of the last 24 years from the Institute for Experimental Pathology at the University of Iceland in Keldur, Iceland laboratory were evaluated to confirm the absence of R. equi cases in Iceland. The diagnostic performance of VapA-specific IgG and its subclasses was evaluated using banked serum samples from pneumonic (n = 21) and healthy foals (n = 80). To evaluate each IgG assay, a cut-off value was selected based on receiver operating characteristic curve analysis and used to calculate sensitivity and specificity. The intra- and interassay coefficients of variation were calculated for each ELISA. Using sera from Iceland, where R. equi infection has not been reported, the VapA-specific IgG ELISA differentiated exposed from nonexposed horses. When used to identify infected foals, VapA-specific IgG, IgGa and IgGb had no diagnostic value. In contrast, IgG(T) had high sensitivity and specificity. Horses from Iceland are not exposed to VapA(+) R. equi and can serve as negative controls. VapA-specific IgG subclasses, with the exception of IgG(T), are poor predictors of disease. Further

  16. [Evaluation of the IgG anti-Toxoplasma response and its avidity by western-blot in HIV-patients].

    Science.gov (United States)

    Cristina Sarmiento, María; Gómez Marín, Jorge Enrique; Castaño Osorio, Jhon Carlos

    2005-01-01

    The IgG anti-Toxoplasma western blot technique was used in 25 HIV-cases and 8 control sera from patients without HIV infection aimed at evaluating the humoral response in these patients. They were divided into 3 groups: 14 HIV positive cases with cerebral toxoplasmosis and IgG anti-Toxoplasma serological titers, 11 HIV positive cases without cerebral toxoplasmosis and with IgG anti-Toxoplasma titers, and 8 HIV negative patients with IgG anti-Toxoplasma titers. It was found that the higher the IgG anti-Toxoplasma serum titers are, the greater the number of bands in the western-blot is. The intensity of the bands measured by densitometry varied significantly for proteins of 66 and 31 kDa. According to the results, these proteins are of interest to evaluate their role in the reactivation of toxoplasmosis in HIV patients.

  17. Análisis inmunoenzimático (ELISA para determinar niveles de IgG anti Bothrops atrox en accidente ofídico Immunoenzymatic determination of IgG anti bothrops atrox levels after snake bites

    Directory of Open Access Journals (Sweden)

    John J. Estrada

    1993-01-01

    Full Text Available Se desarrolló un método de inmunización de conejos con veneno de Bothrops atrox con el fin de preparar antisueros y estandarizar un inmunoanálisis (ELISA para medir niveles de IgG en pacientes con accidente ofídico. La respuesta Inmune de los conejos se siguió por inmunodifusión en doble dimensión (Ouchterlony e inmunoelectroforesis, demostrando la presencia de bandas nítidas desde el día 60 y en todas las sangrias posteriores; se comprobó que hay variabilidad individual en su respuesta inmune. El ELISA para detección de IgG humana anti B. atrox en los indígenas del Chocó fue una prueba simple y sensible (83.3% pero inespecífica por las reacciones cruzadas en individuos que habían sufrido accidentes por B. nasutus. La técnica para detectar IgG equina anti B. atrox en pacientes tratados con antiveneno fue tambIén simple y muy sensible. We developed an immunization method for the production of rabbit antisera against Bothrops atroxvenoms. An enzyme-Ilnked assay (ELISA was standardized in order to measure IgG levels after snake bites. The immune response of rabbits, as determined by Ouchterlony and immunoelectrophoresis techniques, revealed bands of precipitation from the sixtieth day on. Individual variability in the immune response of rabbits was demonstrated. For the measurement of IgG levels In Indians from the Department of Choco (Colombia, ELISA proved to be a sensitive (83.3% and simple but not an specific procedure, since there were cross-reactions in those previously bitten by B. nasutus. ELISA was also simple and sensitive (100% for the determination of equine anti B. atrox IgG antibodies in patients treated with antivenom

  18. Avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii: immunological marker for acute recent toxoplasmosis Avidez de anticorpos IgG anti-antígeno de secreção e excreção de Toxoplasma gondii: marcador imunológico de toxoplasmose aguda recente

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    Patrícia Regina Barboza Araújo

    2008-04-01

    Full Text Available Detection of anti-toxoplasma IgM antibodies has frequently been used as a serological marker for diagnosing recently acquired toxoplasmosis. However, the persistence of these antibodies in some patients has complicated the interpretation of serological results when toxoplasmosis is suspected. The purpose of the present study was to evaluate the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii by means of immunoblot, to establish a profile for acute recent infection in a single serum sample and confirm the presence of residual IgM antibodies obtained in automated assays. When we evaluated the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii by means of immunoblot, we observed phase-specific reactivity, i.e. cases of acute recent toxoplasmosis presented low avidity and cases of non-acute recent toxoplasmosis presented high avidity towards the 30kDa protein fraction, which probably corresponds to the SAG-1 surface antigen. Our results suggest that the avidity of IgG antibodies against excreted/secreted antigens of Toxoplasma gondii is an important immunological marker for distinguishing between recent infections and for determining the presence of residual IgM antibodies obtained from automated assays.A detecção de anticorpos IgM antitoxoplasma tem sido freqüentemente utilizada como marcador sorológico para o diagnóstico de toxoplasmose de aquisição recente. Entretanto, a persistência destes anticorpos em alguns pacientes tem complicado a interpretação dos resultados sorológicos quando a toxoplasmose é suspeitada. A proposta deste trabalho foi avaliar a avidez de anticorpos IgG contra antígenos de secreção e excreção de Toxoplasma gondii por immunoblot, para estabelecer um perfil de infecção recente aguda em uma única amostra de soro e confirmar a presença de anticorpos IgM residuais obtidos nos testes automatizados. Quando a avidez de anticorpos IgG contra ant

  19. Biotin-avidin sandwich elisa with specific human isotypes IgG1 and IgG4 for Culicidae mosquito blood meal identification from an epizootic yellow fever area in Brazil

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    AM Marassá

    2009-01-01

    Full Text Available With a view toward investigating the feeding behavior of Culicidae mosquitoes from an area of epizootic yellow fever transmission in the municipalities of Garruchos and Santo Antônio das Missões, Rio Grande do Sul State, Brazil, specimens were collected by aspiration from September 2005 to April 2007. The engorged females were submitted to blood meal identification by enzyme-linked immunosorbent assay (ELISA. A total of 142 blood-engorged samples were examined for human or monkey blood through species-specific IgG. Additional tests for specificity utilizing isotypes IgG1 and IgG4 of human monoclonal antibodies showed that only anti-human IgG1 was effective in recognizing blood meals of human origin. The results indicated a significant difference (p = 0.027 in detection patterns in samples of Haemagogus leucocelaenus recorded from human blood meals at Santo Antônio das Missões, which suggests some degree of exposure, since it was an area where epizootic outbreaks have been reported.

  20. Evaluation of an Anti-rPA IgG ELISA for Measuring the Antibody Response in Mice

    National Research Council Canada - National Science Library

    Little, S

    2004-01-01

    A recombinant protective antigen (rPA)-based enzyme-linked immunosorbent assay (ELISA) was developed to measure the serological response of female A/J mice after inoculation with the new rPA-based anthrax vaccine...

  1. Detection of anti-dsDNA by IgG ELISA test using two different sources of antigens: calf thymus versus E.coli

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    Mohammadi M

    2009-04-01

    Full Text Available "nBackground: Anti-dsDNA antibodies frequently found in the sera Systemic Lupus Erythematosus patients, particularly in active disease stage. Nowadays exploit different eukaryotic and prokaryotic dsDNA as antigen source and different reagents as binder. The aim of this study to compared two dsDNA different sources and tow different kinds of reagents for binder in ELISA test. "nMethods: In this study bacterial genomic DNA from E.coli (ATCC 25922 and genomic DNA from calf thymus extracted with high purity and were used as antigens for IgG anti-dsDNA detection by ELISA. To coat dsDNA in microtiter wells, tow different kinds of reagents including methylated -BSA and poly-l-lysine (for pre-coating are used. Sera from systemic lupus erythematosus patients and from normal blood donors are used to assess sensitivity and specificity of our ELISA test in compared with IF test and commercial kits. "nResults: Our results displayed pre-coating of microtiter plates with methylated -BSA reduce nonspecific binding reaction and the relative sensitivity and specificity of ELISA increased when calf thymus DNA is employed as antigenic source in compared with IF test and commercial kits 80%, 88% and 100%, 98% respectively, but when E.coli DNA is used 73%, 69% and 85%, 79%, respectively. "nConclusion: The genomic DNA from calf thymus is a potentially useful source of antigen for detection of anti-dsDNA by ELISA. Also the use of methylatted- BSA could have an effective role in reducing of nonspecific binding reactions.

  2. Ultrastructural visualization of the transmembranous and cytomatrix-related part of nicotinic acetylcholine receptor of frog motor endplate by means of an immunochemical avidity of IgG for d-tubocurarine.

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    Shigeru Tsuji

    2008-02-01

    Full Text Available In the present study, a fine ultrastructural localization of nicotinic acetylcholine receptor (nAChR was attempted, using d-tubocurarine (d-TC, a quaternary ammonium compound binding to nAChR. The localization was based on the binding avidity of immunoglobulin G (IgG for acetylcholine (ACh and other quaternary ammonium compounds, such as d-TC. d-TC was applied to the frog neuromuscular preparation and caused a blockade of neuromuscular transmission. Then, d-TC was rendered insoluble in situ by silicotungstic acid (STA, a precipitating agent of soluble proteins and quaternary ammonium compounds. After tissue fixation, a normal rabbit serum was applied to the fine precipitate of the insoluble salt of d-TC silicotungstate (quaternary ammonium radical of d-TC to form the immunochemical complex d-TC- rabbit IgG at ACh binding sites. The IgG of the complex was revealed by means of the conventional immunoperoxidase procedure used for ultrastructural localization. Under the electron microscope, fine diaminobenzidine (DAB precipitates appeared as regular rod-like structures oriented to cytoplasmic side of the horizontal part (crest of the postsynaptic membrane (between the junctional folds which is known to be endowed with nAChR. The rod-like precipitates were not observed in the postsynaptic junctional folds which are devoid of nAChR. The distance separating the rods each other was rather constant (12 - 15 nm, while the length of the rods was variable and exceeded the usual length of nAChR. The present work indicates that the rod-like structures, already observed in association with sarcoplasmic side of the postsynaptic membrane, did correspond to the intramembranous and intracytoplasmic part of nAChR and related proteins. These cytochemical results confirm that d-TC binds to ACh binding sites in the pore of nAChR, and raise the question of DAB staining of cytoskeletal proteins related to the nAChR complex.

  3. Correlation between specific IgM levels and percentage IgG-class antibody avidity to Toxoplasma gondii Correlação entre n��veis de IgM específica e percentual de avidez de anticorpos da classe IgG ao Toxoplasma gondii

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    Marcel Leite

    2008-08-01

    Full Text Available Toxoplasmosis is an usually asymptomatic worldwide disseminated infection. In its congenital presentation it may lead to abortion or fetal malformations. Antenatal evaluation is considered of paramount importance to identify seronegative women and allow for prophylaxis. Recent improvements in sensitivity of IgM tests has made IgM detection an extremely protracted acute phase marker, and IgG avidity evaluation test became necessary. Observation has shown that a correlation can be established between IgM levels and avidity percentages, suggesting that frequently the avidity test may not be necessary. In this study we analyzed Toxoplasma gondii IgM levels of 202 samples and their IgG avidity percentages, in order to define specific levels whose IgM quantification could by itself define serodiagnosis and therefore make the avidity evaluation unnecessary. We showed that for IgM levels bellow 2.0 and above 6.0 serodiagnosis of toxoplasmosis could be established without need of IgG avidity test. IgM levels between these two parameters are associated with varying avidity indexes highlighting the importance of its evaluation as a means to confirm toxoplasmosis. Following this demonstration it was possible to avoid the avidity test for 75% of the cases, to reduce the turnaround time and to reduce costs.A Toxoplasmose é uma infecção universal e usualmente assintomática. A forma congênita, entretanto, pode resultar em aborto ou mal formações. Testes sorológicos estão indicados em situações onde há suspeita clínica, e na triagem pré-natal, quando são extremamente importantes para rastrear a infecção e orientar a gestante. O aumento da sensibilidade das técnicas para detecção de IgM, tornou necessário o desenvolvimento de recursos, como a avidez de IgG, visando obter novo marcador de infecção aguda. Embora exista correlação entre níveis de IgM e grau de avidez de IgG, a maioria dos testes de avidez associa-se a níveis baixos de Ig

  4. ELISA detection of IgG antibody against a recombinant major surface antigen (Nc-p43) fragment of Neospora caninum in bovine sera

    Science.gov (United States)

    Ahn, Hye-Jin; Kim, Sera; Kim, Dae-Yong

    2003-01-01

    An ELISA was established to measure bovine IgG directed against the recombinant antigenic determinant of Nc-p43, a major surface antigen of Neospora caninum. In a previous study, two thirds of the C-terminal of the molecule was expressed as a 6 × His tagged protein (Ncp43P) for ELISA using 2/3 of the N-terminal of SAG1 from Toxoplasma gondii as a control (TgSAG1A). Among 852 cattle sera collected from stock farms scattered nation-wide, 103 sera (12.1%) were found to react with Ncp43P positively, but no positive reaction was observed with TgSAG1A. This study shows that Ncp43P could be available as an efficient antigen for the diagnosis of neosporosis in cattle. Furthermore, it together with TgSAG1A, could be useful for the differential diagnosis of N. caninum and T. gondii infections in other mammals. PMID:12972732

  5. Establishment of an antibody avidity test to differentiate vaccinated cattle from those naturally infected with Mycoplasma bovis.

    Science.gov (United States)

    Han, Xiaoxiao; Khan, Farhan Anwar; Zhu, Xifang; Zhang, Rui; Mustafa, Riaz; Hu, Changmin; Chen, Yingyu; Chen, Huanchun; Guo, Aizhen

    2015-01-01

    Mycoplasma bovis is a major pathogen of bovine respiratory disease (BRD) in China and a live attenuated vaccine has recently been developed. This study aimed to establish an IgG avidity test to differentiate between naturally infected and vaccinated animals. An indirect ELISA (iELISA) was first established in the laboratory to detect antibodies specific to M. bovis using whole cell proteins as coating antigens and serum samples from experimentally infected cattle. The specificity and sensitivity of the iELISA was confirmed using a commercial ELISA kit as a reference standard. Both tests showed substantial agreement as indicated by a κ value of 0.78 (95% confidence interval, CI, 0.62, 0.93), and an overall 92.0% (80/87) agreement between the two tests. Based on the laboratory iELISA, a sodium thiocyanate (NaSCN) competitive iELISA was then developed for the detection of IgG avidity, expressed as relative avidity index (AI). Two-hundred and one experimentally immunised and naturally infected animals were used. These comprised 36 immunised calves, 38 negative control calves, 37 naturally infected calves, 87 calves of unknown status, and an additional three immunised calves that were used for a time trial. By testing true positive and negative antisera from either naturally infected or immunised calves, the AI cut-off value was defined as 70.4%. The diagnostic accuracy of the in-house NaSCN competitive iELISA was determined using serum samples collected from the experimental animals. The IgG avidity test demonstrated 96.0% sensitivity (95% CI 80.5%, 99.3%) and 95.8% specificity (95% CI 79.8%, 99.3%), and was successfully established as a valuable first test for differentiating vaccinated animals from those infected with M. bovis. This test may be a useful tool for clarifying the magnitude of M. bovis infection and in assessing the efficacy of vaccination in exposed animal populations. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Leptospirosis serosurvey in bovines from Brazilian Pantanal using IGG ELISA with recombinant protein LipL32 and microscopic agglutination test Sorodiagnóstico de leptospirose em bovinos do Pantanal brasileiro utilizando ELISA IgG com proteína recombinante LipL32 e soroaglutinação microscópica

    Directory of Open Access Journals (Sweden)

    Renata Graça Pinto Tomich

    2007-12-01

    Full Text Available This investigation was carried out in Brazilian Pantanal: region with important biodiversity. This region's climatic conditions, hydrology and geomorphology as well as the existence of great variety of wild species favor the maintenance of the Leptospira in the environment. The aim of this study was to evaluate IgG ELISA with recombinant protein LipL32 in comparison with microscopic agglutination test (MAT and additionally contribute to the knowledge of the distribution of the one of most important worldwide zoonotic infection, assessing the seropositivity of bovine leptospirosis in beef cattle herds of Brazilian Pantanal, an important ecological preserved area, where cattle constitute not only the most important economic resource but also the major activity compatible of the conservation of natural resource of the region. Out of 282 samples of cattle serum analyzed, 143 (50.71% were positive in MAT. The serovar Hardjo (genotypic Hardjoprajitno and Hardjobovis, Wolffi and Ballum showed the largest frequency of reactive samples. In the IgG ELISA rLipL32, 161 samples (57.09% were positive. This result was higher than obtained by MAT (pEste estudo foi realizado no Pantanal brasileiro: região que apresenta importante biodiversidade. As condições de clima, hidrologia e geomorfologia dessa região, bem como a existência de grande variedade de espécies animais silvestres, favorecem a manutenção da Leptospira no meio ambiente. O objetivo desse estudo foi avaliar o ELISA IgG com proteína recombinante LipL32 em comparação com a soroaglutinação microscópica (SAM para o diagnóstico sorológico de Leptospira. Adicionalmente, contribuir para o conhecimento da distribuição da leptospirose bovina, uma das mais importantes zoonoses mundialmente distribuída. Foi avaliada a soropositividade para essa bactéria em rebanhos bovinos de corte da região do Pantanal, uma área onde o bovino constitui não apenas o recurso econômico mais importante

  7. Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies.

    Science.gov (United States)

    Sonmez, Cemile; Coplu, Nilay; Gozalan, Aysegul; Akin, Lutfu; Esen, Berrin

    2017-06-01

    Detection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml. A total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01-0.1IU/ml, 0.1-1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml). A statistically significant correlation (r 2 =0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%. In-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected. Copyright © 2017. Published by Elsevier B.V.

  8. Two years' performance of an in-house ELISA for diagnosis of Legionnaires' disease: detection of specific IgM and IgG antibodies against Legionella pneumophila serogroup 1, 3 and 6 in human serum.

    Science.gov (United States)

    Elverdal, P L; Jørgensen, C S; Krogfelt, K A; Uldum, S A

    2013-08-01

    The aim of this study was to evaluate the performance of an in-house ELISA for the diagnosis of Legionnaires' disease (LD) by detection of IgM and IgG antibodies against Legionella (L.) pneumophila serogroups (sg) 1, 3 and 6. The evaluation was done throughout a two-year period in a diagnostic routine laboratory. Furthermore, the sensitivity of four different methods, the in-house L. pneumophila antibody test (ELISA), the urinary antigen test (Binax® EIA), an in-house PCR and culture, both alone and in combination was evaluated. From 2008 to 2010, 12,158 serum samples from 10,503 patients were analysed. During the same period, 361 cases of laboratory-confirmed LD cases were recorded in Denmark, but of these only 113 had a serum sample examined. The positive predictive value of the in-house ELISA was calculated to be 12.8 and the negative predictive value was 99.6, using only the confirmed LD cases as true positives. The sensitivity of the in-house ELISA for the detection of IgM and IgG antibodies in the confirmed LD cases was 61% and 36%, respectively. By combining the two ELISA assays the sensitivity increased to 66%. The sensitivity of the Legionella urinary antigen test (Binax® EIA) was 63%, of the in-house PCR 87% and of culture 69%. When all the different methods were combined, a higher sensitivity was calculated--for in-house ELISA (IgM+IgG) and Binax® EIA 91%, in-house ELISA (IgM+IgG) and in-house PCR 93%, in-house ELISA (IgM+IgG) and culture 93%, Binax® EIA and in-house PCR 79%, Binax® EIA and culture 68% and in-house PCR and culture 94%. This study confirms that the detection of IgG and IgM antibodies by ELISA is an important diagnostic tool, also during the initial phase of the disease. Furthermore, we showed that LD in Denmark with or without serum samples collected exhibits the same age and sex distribution and epidemiology, as in the rest of Europe, i.e., mostly men are infected, infections are mostly community acquired, followed by infection from

  9. Discrepancies between a new highly sensitive Toxoplasma gondii ELISA assay and other reagents: interest of Toxo IgG Western blot.

    Science.gov (United States)

    Leslé, F; Touafek, F; Fekkar, A; Mazier, D; Paris, L

    2011-10-01

    Immunodiagnostic assays are commonly used to screen for maternal toxoplasmic seroconversion during pregnancy. The introduction to the market of a new highly sensitive IgG assay, the Elecsys Toxo IgG test, has resulted in discrepancy issues with other immunoassays because of a lack of standardisation. Western blot appears to be a good alternative gold standard to the dye test, as the latter is not routinely available. For the present prospective study, we compared the analytical performances of two immunoassays, Elecsys Toxo IgG (Roche Diagnostics) and Platelia Toxo IgG (Bio-Rad, Marnes la Coquette, France), to Toxo II IgG Western blot (LDBio, Lyon, France) using 231 consecutive sera with low or equivocal IgG titres. Of these 231 sera, 213 presented discrepancies, which showed the importance of a confirmation test. Of the Elecsys Toxo IgG-positive results, 100% were confirmed by the Western blot with a positive threshold of 30 IU/ml for Elecsys; in the equivocal area (1-30 IU/ml), Western blot is negative in 54% of cases. Our results suggest that the lower diagnostic cut-off of Platelia Toxo IgG should be further reduced. Our study indirectly confirms that monitoring, especially for pregnant women, must be done in the same laboratory using the same technique. The ability to diagnose very early seroconversion using Western blot merits further study.

  10. Anti-neosporal IgG and IgE antibodies in canine neosporosis.

    Science.gov (United States)

    Jesus, E E V; Almeida, M A O; Atta, A M

    2007-01-01

    Neospora caninum infection provokes neurological disorders, recurrent abortion and death in dogs and cattle. Dogs are both intermediate and definitive host of N. caninum. Thus, the development of sensitive and specific immunoassays to diagnose canine neosporosis is essential to control this disease. This work investigated serum anti-neosporal IgG and IgE antibodies in 140 dogs represented by 30 healthy animals (group I), 11 dogs showing acute N. caninum infection (group II), 50 urban dogs with serological evidence of canine neosporosis in indirect fluorescent antibody test (IFAT) (group III) and 49 urban dogs without clinical and laboratory evidences of neosporosis (group IV). Enzyme-linked immunosorbent assay (ELISA) and western immunoblotting, both using a soluble N. caninum tachyzoite antigen (SNA), investigated these two isotypes of antibodies, while a Urea-ELISA measured the avidity of the IgG antibodies. Anti-Toxoplasma gondii IgG antibodies were also investigated in the animals. Anti-neosporal IgG was found in all animals from groups II and III, whereas 32.7% (16/49) of dogs from group IV were reactive. IgG antibodies of low avidity were demonstrated in dogs from group II (median 35.3%), while animals from groups III and IV had IgG antibodies of high avidity (medians of 61.5% and 61.7% respectively). IgE antibodies were found in four (13.3%) and five (16.6%) dogs from groups III and IV respectively. Dogs presenting acute infection (group II) or chronic infection (group III) had IgG antibodies to several neosporal antigens, mainly of 29-30 and 35 kDa, while 13 of 16 dogs from group IV recognized antigens from 14 to 170 kDa. Antibodies to T. gondii were detected in 36 of 50 (72%) sera from group III and 25 of 49 (51%) sera from group IV. We concluded that IgG-ELISA and Urea-ELISA with SNA may substitute for IFAT in both laboratory routine and epidemiological studies of canine neosporosis.

  11. Enzyme linked immunosorbent assay (ELISA for the detection of IgG, IgM, IgE and IgA against Cysticercus cellulosae in cerebrospinal fluid of patients with neurocysticercosis

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    Odashima Newton Satoru

    2002-01-01

    Full Text Available The objective of this study was to analyze different immunoglobulins classes (IgG, IgM, IgE and IgA against Cysticercus cellulosae in the cerebrospinal fluid (CSF, through enzyme linked immunosorbent assay (ELISA, correlating them to clinical and tomographic profiles in patients with neurocysticercosis (NCC. Eighty-five specimens of CSF were obtained from 43 cases with NCC (26 with the active form and 17 with the inactive form and from 42 patients with other neurological diseases. The inactive form of NCC presented a profile in CSF similar to the group without NCC. The active form of NCC presented elevation of specific immunoglobulins (IgG, IgM, IgE, and IgA in decreasing order, with the highest values being detected among the cases with intraventricular cysts, or with inflammation signs in CSF or in those with multiple clinical manifestations. The highest sensitivity and specificity were obtained with ELISA-IgG (88.5% and 93.2%, respectively. This study confirmed the importance of ELISA in the immunologic diagnosis of NCC.

  12. Toxoplasma-IgM and IgG-avidity in single samples from areas with a high infection rate can determine the risk of mother-to-child transmission Avidez de IgM e IgG de Toxoplasma em amostras de áreas com alta taxa de infecção pode determinar risco de transmissão materno-fetal

    Directory of Open Access Journals (Sweden)

    Myrian Morussi Reis

    2006-04-01

    Full Text Available Anti-Toxoplasma IgG-avidity was determined in 168 serum samples from IgG- and IgM-positive pregnant women at various times during pregnancy, in order to evaluate the predictive value for risk of mother-to-child transmission in a single sample, taking the limitations of conventional serology into account. The neonatal IgM was considered the serologic marker of transmission. Fluorometric tests for IgG, IgM (immunocapture and IgG-avidity were performed. Fifty-one of the 128 pregnant women tested gave birth in the hospital and neonatal IgM was obtained. The results showed 32 (62.75% pregnant women having high avidity, IgM indexes between 0.6 and 2.4, and no infected newborn. Nineteen (37.25% had low or inconclusive avidity, IgM indexes between 0.6 and 11.9, and five infected newborns and one stillbirth. In two infected newborns and the stillbirth maternal IgM indexes were low and in one infected newborn the only maternal parameter that suggested fetal risk was IgG-avidity. In the present study, IgG-avidity performed in single samples from positive IgM pregnant women helped to determine the risk of transmission at any time during pregnancy, especially when the indexes of the two tests were analysed with respect to gestational age. This model may be less expensive in developing countries where there is a high prevalence of infection than the follow-up of susceptible mothers until childbirth with monthly serology, and it creates a new perspective for the diagnosis of congenital toxoplasmosis.A avidez de IgG anti-Toxoplasma foi realizada em 168 amostras IgG e IgM positivas de gestantes, coletadas em qualquer período da gestação, para avaliar o valor preditivo do risco de transmissão materno-fetal em amostra única, considerando as limitações da sorologia convencional. A IgM neonatal foi considerada o marcador sorológico de transmissão. Testes fluorométricos foram realizados para IgG, IgM (imunocaptura e avidez de IgG. Cinqüenta e uma das 128

  13. Estandarización en Colombia de una prueba ELISA para la evaluación de los niveles séricos de anticuerpos IgG contra diez serotipos de Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Lucía Carolina Leal-Esteban

    2012-03-01

    Conclusión. Esta prueba ELISA cuantifica de forma confiable los niveles de IgG sérica contra diez serotipos de S. pneumoniae y, de acuerdo con los resultados obtenidos en individuos sanos de nuestra población, en este trabajo se validan los parámetros internacionales para considerar adecuada la respuesta a la vacuna 23-valente contra este microorganismo.   DOI: http://dx.doi.org/10.7705/biomedica.v32i1.393

  14. Comparison of immunofluorescence and enzyme-linked immunosorbent assay and immunoglobulin G avidity techniques for screening of anti: Toxoplasma antibodies among single serum sample pregnant women in Tabriz, Iran

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    Mehrangiz Rajaii

    2015-01-01

    Full Text Available Background: Congenital toxoplasmosis is that pregnant women acquire the infection during gestation; diagnosis of the acute infection during pregnancy is a complex subject of maternal toxoplasmosis. Thus, the presence of immunoglobulin G (IgG and/or IgM Toxoplasma antibodies in a single serum sample drawn during gestation cannot be used to define whether the infection was recently acquired or chronic. Materials and Methods: At this cross-sectional descriptive study, sera of 391 pregnant women examined and compared. They were in an age range of 21-35 years, referred by gynecologists and infectious disease specialists, during March 2012-April 2013. They have referred, 215 (54.98%, 102 (26%, 74 (18.92% in the first, second and third trimesters of gestation, respectively. For each of them, a questionnaire was completed and serum samples were prepared in an equal condition, examined according to the procedures of indirect immunofluorescence (IIF, enzyme-linked immunosorbent assay (ELISA and IgG Avidity techniques. Results: We have found 111 (28.38% seronegative and 280 (71.61% seropositive cases by IIF and 124 (31.70% seronegative, 267 (68.28% seropositive cases by ELISA. The IgG avidity test confirmed 45 (69.23% and 7 (10.76% doubtful cases of IgM test in IIF and ELISA techniques. Conclusions: This study highlights how to manage pregnant women with toxoplasmosis, especially in a single serum sample condition.

  15. Validation of an ELISA for the concurrent detection of total antibodies (IgM and IgG to Rift Valley fever virus

    Directory of Open Access Journals (Sweden)

    Charlotte E. Ellis

    2014-05-01

    Full Text Available Rift Valley fever virus (RVFV infects humans and livestock, causing haemorrhaging andabortions in animals. Three major RVF epizootics have occurred in South Africa since the1950s and the outbreak in 2010 had a mortality rate of 10.7% in humans. Accurate and earlydetection is therefore essential for management of this zoonotic disease. Enzyme-linkedimmunosorbent assays (ELISAs have been developed for the detection of either IgM or IgGantibodies to RVFV in animal sera. In this study, data are presented on the validation of adouble-antigen ELISA for the simultaneous detection of both classes of antibodies to RVFV ina single test. ELISA plates were coated with a recombinant nucleoprotein. The nucleoprotein,conjugated to horseradish peroxidase, was used as the detecting reagent. A total of 534 serafrom sheep and cattle were used in the validation. The sheep sera were collected during a RVFpathogenesis study at the Agricultural Research Council (ARC – Onderstepoort VeterinaryInstitute and the cattle sera were collected during an outbreak of RVF in 2008 at the ARC –Animal Production Institute in Irene, Pretoria. The ELISA had a diagnostic sensitivity of 98.4%and a specificity of 100% when compared to a commercial cELISA. This convenient and fastassay is suitable for use in serological surveys or monitoring immune responses in vaccinatedanimals.

  16. Development of ELISAs for quantification of HMFG1-specific human anti-mouse IgG and IgM antibodies.

    Science.gov (United States)

    Oei, A L M; Boerman, O C; Geurts-Moespot, A; van Eerd, J E; van Tienoven, D; Courtenay-Luck, N; Thomas, C M G; Massuger, L F A G; Sweep, F C G J

    2007-01-01

    The aim of this study was to develop and validate ELISAs for quantification of HAMA-IgM and HAMA-IgG in serum of patients with ovarian cancer who enrolled in a large international randomized phase III trial of intraperitoneal Yttrium-90-labeled HMFG1 murine monoclonal antibody therapy. The capture antibody of these 2 assays was the murine antibody HMFG1, while mouse anti-human IgM-HRP or mouse anti-human IgG(Fc)-HRP served as tracer antibodies. A pool of HAMA-positive serum samples was used to prepare a series of assay standards and another pool served as reference preparation. The analytical sensitivity of the HAMA-IgM assay was 2.5 arbitrary units per mL (AU/mL) and 4.7 AU/mL for the HAMA-IgG ELISA. Diluted serum samples showed good parallelism with the HAMA-IgM and HAMA-IgG standard dose-response curves. Within-assay coefficient of variation was 7.5% for HAMA-IgM and 6.5% for HAMA-IgG. Between-assay variation was 14.2% for HAMA-IgM and 15.3% for HAMA-IgG. The developed HAMA-IgM and HAMA-IgG ELISAs show satisfactory reliability criteria (sensitivity, parallelism and precision) and are suitable for monitoring of HAMA-IgM and HAMA-IgG responses in ovarian cancer patients. These ELISAs will be used to monitor the development of HAMAs in patients who received radioimmunotherapy with murine HMFG1.

  17. The blocking activity of birch pollen-specific immunotherapy-induced IgG4 is not qualitatively superior to that of other IgG subclasses

    DEFF Research Database (Denmark)

    Ejrnaes, Anne M; Bødtger, Uffe; Larsen, Jørgen N

    2004-01-01

    blocking activity was found in the purified IgG4 fraction. There was no significant difference in the binding avidities (1/K(d)) measured in the two IgG fractions. Thus, it appears that SIT-induced specific IgG4 contributes to the IgG blocking of allergen binding to IgE in a simple quantitative manner...

  18. Immune response to pneumococcal polysaccharides 4 and 14 in elderly and young adults. I Antibody concentrations, avidity and functional activity

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    Carlone George M

    2005-06-01

    Full Text Available Abstract Streptococcus pneumoniae is a serious worldwide pathogen and the focus of numerous vaccine development projects. Currently the most widely accepted surrogate marker for evaluating the efficacy of a given vaccine is to utilize ELISA. Measurement of antibody concentration by ELISA without reduction in cross-reactive antibodies causes an overestimation of antibody concentration and therefore protection, this is most notable in the aged, an at risk group for this infection. We compared the immune response to the pneumococcal polysaccharides (PPS 4 and 14 of 20 young to 20 elderly adults. Pre-and post-vaccination IgG antibody concentrations and antibody avidity against PPS4 and PPS14 were measured using two different enzyme-linked immunosorbant assay (ELISA absorption protocols. All sera were pre-absorbed with either cell-wall polysaccharide (CPS, or CPS and serotype 22F polysaccharide. Pre- and post-vaccination IgG antibody concentrations for serotype 4, but not 14, were significantly lowered with the additional absorption with serotype 22F polysaccharide in both age groups. Young and elderly demonstrated a significant increase from pre- to post-immunization antibody concentration, using either absorption method; and opsonophagocytic antibody titers in response to both PPS4 and PPS14. The correlation coefficients between ELISA and opsonophagocytic assays were improved by additional absorption with serotype 22F in response to serotype 4, but not serotype 14 in all age groups. Opsonophagocytic antibody titers in a sub-group of elderly (>77 years of age were significantly lower than the opsonophagocytic antibody concentrations in young adults. These results suggest the importance of eliminating cross-reactive antibodies from ELISA measurements by absorption of serum and an age-related impairment in the antibody response to pneumococcal polysaccharides.

  19. Validación de un ELISA para la cuantificación de IgG antipolisacárido capsular de Neisseria meningitidis serogrupo C en sueros de ratones.

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    Maribel Cuello

    2001-12-01

    Full Text Available Se desarrolló un ensayo inmunoenzimático de fase sólida (ELISA indirecto para cuantificar anticuerpos IgG específicos antipolisacárido C en ratón, utilizando un prerrecubrimiento con Poli-L-lisina y luego el polisacárido capsular de Neisseria meningitidis serogrupo C (Instituto Finlay, La Habana, Cuba, para evaluar la respuesta inmune contra este componente en candidatos vacunales en estudios preclínicos. Como conjugado se utilizó anti-IgG ratón conjugado a fosfatasa alcalina, el cual se une a los anticuerpos antipolisacárido C producidos en ratones. La reacción es evidenciada por la degradación del sustrato p-nitrofenilfosfato. La detectibilidad del ensayo fue de 123,74 U/mL y la especificidad fue alta. La precisión interensayo, intraensayo y total, así como las desviaciones de la recuperación, linealidad y paralelismo no sobrepasaron el 10%. El ELISA permitió cuantificar los anticuerpos antipolisacárido C inducidos en ratones tanto por candidatos vacunales conjugados, como por la vacuna VA-MENGOC-BC y el polisacárido C sin conjugar.

  20. Padronização de ensaio imunoenzimático para pesquisa de anticorpos das classes IgM e IgG anti-Toxoplasma gondii e comparação com a técnica de imunofluorescência indireta Standardization of enzyme-linked immunosorbent assay ELISA to detect anti-Toxoplasma gondii IgM and IgG antibodies, and comparison with the indirect immunofluorescence technique

    Directory of Open Access Journals (Sweden)

    Cláudia Maria Antunes Uchôa

    1999-12-01

    Full Text Available A sorologia tem sido o método de escolha para o diagnóstico da toxoplasmose. Devido a isto, padronizamos um ensaio imunoenzimático (ELISA e comparamos seus resultados com a técnica de imunofluorescência indireta (IFI. A técnica padronizada apresentou na pesquisa de IgG sensibilidade (S de 96,7% e especificidade (E de 75%, com valor de predição de positividade (VPP de 83,3% e de negatividade (VPN de 94,7%, com uma concordância ajustada (K de 73,5%. A IFI apresentou S de 83,8%, E de 79,1% com VPP de 83,8 % e VPN de 79,1% com K de 63%. A concordância bruta entre os dois testes (ELISA/IFI foi de 88,3% para pesquisa de IgG e de 81,5% para pesquisa de IgM, sendo o K de 70,8% para IgG e de 1,3% para IgM, sendo o índice de correlação (r de 0,556 para IgG e de -0,023 para IgM. Podemos concluir que a ELISA-IgG padronizada é indicada nos processos de triagem sorológica, sendo a ELISA-IgM desaconselhada uma vez que apresentou baixos índices de concordância ajustada com a técnica de referência, sugerindo pouca confiabilidade dos resultados.Serology has been the most popular method to diagnose toxoplasmosis. Accordingly, this study standardizes an enzyme-linked immunosorbent assay (ELISA and compares its results with the IFI technique. In the IgG detection test, the standardized technique presented a sensibility (S of 96.77%, a specificity (SP of 75%, with a positive predictive value (PPV of 83.33%, a negative predictive value (NPV of 94.74%, and an adjusted concordance (K of 73.50%. The IFI exhibited 83.87% for S, 79.16% for SP, 83.81% for PPV, 79.16% for NPV, and 63% for K. The rough concordance between these two tests (ELISA/IFI was 88.35% for the IgG detection test and 81.55% for the IgM detection test. K was 70.82% and 1.31% for IgG and IgM, respectively, the correlation index (r being 0.556 for IgG and -0.023 for IgM. We can conclude that standardized ELISA-IgG is indicated in serologic selection processes, whereas the ELISA-IgM is

  1. Toxoplasmose em gestantes de Araraquara/SP: análise da utilização do teste de avidez de IgG anti-Toxoplasma na rotina do pré-natal = Toxoplasmosis in pregnant women from Araraquara/SP: analysis of Toxoplasma-specific IgG avidity test utilization in the prenatal care routine

    Directory of Open Access Journals (Sweden)

    Isabel, Thais F.

    2007-01-01

    Conclusões: O tratamento antiparasitário foi prescrito para algumas gestantes mesmo na presença de uma taxa de avidez de IgG anti-T. gondii alta no primeiro trimestre da gestação, mostrando que o teste de avidez não foi corretamente interpretado e utilizado na prática clínica

  2. Direct ELISA.

    Science.gov (United States)

    Lin, Alice V

    2015-01-01

    First described by Engvall and Perlmann, the enzyme-linked immunosorbent assay (ELISA) is a rapid and sensitive method for detection and quantitation of an antigen using an enzyme-labeled antibody. Besides routine laboratory usage, ELISA has been utilized in medical field and food industry as diagnostic and quality control tools. Traditionally performed in 96-well or 384-well polystyrene plates, the technology has expanded to other platforms with increase in automation. Depending on the antigen epitope and availability of specific antibody, there are variations in ELISA setup. The four basic formats are direct, indirect, sandwich, and competitive ELISAs. Direct ELISA is the simplest format requiring an antigen and an enzyme-conjugated antibody specific to the antigen. This chapter describes the individual steps for detection of a plate-bound antigen using a horseradish peroxidase (HRP)-conjugated antibody and luminol-based enhanced chemiluminescence (ECL) substrate. The methodological approach to optimize the assay by chessboard titration is also provided.

  3. Microsatellite typing and avidity analysis suggest a common source of infection in herds with epidemic Neospora caninum-associated bovine abortion.

    Science.gov (United States)

    Basso, W; Schares, S; Minke, L; Bärwald, A; Maksimov, A; Peters, M; Schulze, C; Müller, M; Conraths, F J; Schares, G

    2010-10-11

    Neosporosis is an important cause of reproductive failure in cattle worldwide. Two different abortion patterns associated with Neospora caninum infection have been observed in cattle herds: endemic and epidemic abortion outbreaks. The endemic pattern is characterized by an abortion problem in a herd persisting for several months or years, and is assumed to be caused by reactivation of a chronic infection. In epidemic outbreaks, abortions concentrate within a short period of time, most likely due to a recent point source exposure of naïve animals to N. caninum. The aim of the study was to characterize five N. caninum-associated epidemic abortion outbreaks in Germany by serological and molecular techniques, including a p38-avidity-ELISA and typing of N. caninum in clinical samples by multilocus-microsatellite analysis. DNA extracts from the brain of 18 N. caninum infected fetuses from epidemic abortion outbreaks were characterized using 10 N. caninum-microsatellite markers. Nested-PCR protocols were developed to amplify the marker regions MS1B, MS3, MS5, MS6A, MS6B, MS7, MS12 and MS21 from clinical samples for subsequent analysis by capillary electrophoresis. Microsatellites MS2 and MS10 were analyzed by previously reported sequencing techniques. Most dams which had aborted showed a low-avidity IgG response to the N. caninum p38-antigen, and in three of the five studied herds, the majority of the dams at risk, which had not aborted, had also low-avidity responses suggesting that infection with N. caninum had recently occurred in most animals. A common microsatellite pattern prevailed in all fetuses from each individual epidemic outbreak. This pattern was unique for each herd. Although the number of epidemic abortion outbreaks analyzed was limited, the observation of a common microsatellite pattern, accompanied by a low-avidity IgG response against N. caninum in the dams, supports the hypothesis of a recent infection from a common point source. The genetic diversity

  4. Avidity of Antibodies against HSV-2 and Risk to Neonatal Transmission among Mexican Pregnant Women

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    Antonia Herrera-Ortiz

    2013-01-01

    Full Text Available Objective. To determine HSV-2 seroprevalence, risk factors, and antibody avidity among a sample of Mexican pregnant women. Material and Methods. The avidity test was standardized with different urea concentrations and incubation times; the cut-off point was calculated to determine the low avidity (early infection. IgG antibodies against HSV-2 were detected from pregnant and postpartum women from Morelos, Mexico, and the avidity test was performed to positive samples. Multivariate regression logistic analysis was employed to evaluate demographic and sexual behavior characteristics associated with HSV-2 infection. Results. HSV-2 seroprevalence among Mexican women analyzed was 14.5% (333/2300, demographic factors (location of General Hospital, age, education level, and civil status, and risky sexual behaviors (STI self-report and number of sexual partners during last year were associated with HSV-2 infection. Seventeen women were detected with low avidity antibodies (early infection with a cut-off point of 66.1%. Conclusions. HSV-2 infection was common among this group of women from Mexico; the avidity test detected women with recent infections, and these women were more likely to transmit HSV-2 to their neonates. Neonatal herpes has no epidemiological surveillance, the disease could be overlooked, and so more studies are needed to estimate the magnitude of neonatal infection.

  5. Hu and Yo antibodies have heterogeneous avidity.

    Science.gov (United States)

    Totland, Cecilie; Aarseth, Jan; Vedeler, Christian

    2007-04-01

    Onconeural antibodies such as anti-Hu and anti-Yo may be important in the pathogenesis of paraneoplastic neurological syndromes. The avidity of these antibodies is not known. In this study, we compared the avidity of Hu and Yo antibodies both at single time points and over a time range of 2 months to 6 years. The avidity of Yo and Hu antibodies differed among the patients, but anti-Yo generally had higher avidity than anti-Hu. Whether Yo antibodies are more pathogenic than Hu antibodies are presently unknown.

  6. Malaria serology: performance of six Plasmodium falciparum antigen extracts and of three ways of determining serum titers in IgG and IgM-ELISA Sorologia da malária: desempenho de seis diferentes extratos antigênicos de Plasmodium falciparum e de três métodos de determinação de títulos de soros no teste ELISA-IgG e IgM

    Directory of Open Access Journals (Sweden)

    Maria Carmen Arroyo Sanchez

    1993-12-01

    Full Text Available The study evaluated six Plasmodium falciparum antigen extracts to be used in the IgG and IgM enzyme-linked immunosorbent assays (ELISA, for malaria diagnosis and epidemiological studies. Results obtained with eighteen positive and nine negative control sera indicated that there were statistically significant differences among these antigen extracts (Multifactor ANOVA, pEste estudo avaliou seis extratos antigênicos de Plasmodium falciparum para utilização no diagnóstico da malária e em estudos epidemiológicos, empregando o teste "enzyme-linked immunosorbent assay" (ELISA para a pesquisa de anticorpos das classes IgG e IgM. Os resultados obtidos empregando dezoito soros padrão positivo e nove padrão negativo, indicaram que houve diferenças estatisticamente significativas entre os extratos antigênicos estudados (Multifactor ANOVA, p <0,0001. Extratos antigênicos preparados com uréia, desoxicolato de sódio e Zwittergent forneceram melhores resultados que os demais e se comportaram de modo bastante semelhante, para a detecção de anticorpos IgG. Extratos antigenicos de uréia, alcalino e de desoxicolato de sódio foram os melhores na detecção de anticorpos IgM. Devido à relação linear obtida entre as densidades ópticas (ou os log10 respectivos e os log10 das diluições, com uma inclinação constante, os títulos dos soros foram determinados através de duas equações diferentes, que necessitam apenas de uma diluição do soro. Para anticorpos IgM, a expressão que emprega log10 da densidade óptica forneceu resultados que se correlacionaram melhor com a titulação (95% Bon-ferroni. Para anticorpos IgG, as diferenças entre resultados não foram significativas. A avaliação da reprodutibilidade dos títulos e das partidas de antígenos forneceu resultados satisfatórios.

  7. Elisa Valero

    OpenAIRE

    Valero Ramos, Elisa

    2009-01-01

    MENOR O IGUAL QUÉ. Los arquitectos, habitualmente aficionados a la matemática sencilla, a los enteros más que a los quebrados, a los módulos, las series, los ejes, las simetrías y las transparencias, admiramos ese oximorón miesiano de ¿menos es más¿ que es, también, el sumum del expresionismo elemental. Y tal vez la arquitecta Elisa Valero, cuya obra autodenominada menor tengo el honor de glosar con estas letras, milita también en este orden de ideas. Educada a partes iguales entre e...

  8. Avidity of onconeural antibodies is of clinical relevance.

    Science.gov (United States)

    Totland, Cecilie; Ying, Ming; Haugen, Mette; Mazengia, Kibret; Storstein, Anette; Aarseth, Jan; Martinez, Aurora; Vedeler, Christian

    2013-08-01

    Onconeural antibodies are important in the detection of paraneoplastic neurological syndromes (PNS). The avidity of Hu, Yo, and CRMP5 antibodies from 100 patients was determined by immunoprecipitation (IP), and 13 of the Yo positive sera were also tested by surface plasmon resonance (SPR). There was a significant association between the results from IP and SPR. Yo antibodies had higher avidity than Hu and CRMP5 antibodies, and both high- and low-avidity antibodies were associated with tumors and PNS. High-avidity Yo antibodies were mainly associated with ovarian cancer, whereas high-avidity Hu and CRMP5 antibodies were mainly associated with small-cell lung cancer. Low-avidity CRMP5 and Yo antibodies were less often detected by a commercial line blot than high-avidity antibodies. The failure to detect low-avidity onconeural antibodies may result in under diagnosis of PNS.

  9. Influence of maternal vaccination against diphtheria, tetanus, and pertussis on the avidity of infant antibody responses to a pertussis containing vaccine in Belgium.

    Science.gov (United States)

    Caboré, Raïssa Nadège; Maertens, Kirsten; Dobly, Alexandre; Leuridan, Elke; Van Damme, Pierre; Huygen, Kris

    2017-10-03

    Maternal antibodies induced by vaccination during pregnancy cross the placental barrier and can close the susceptibility gap to pertussis in young infants up to the start of primary immunization. As not only the quantity but also the quality of circulating antibodies is important for protection, we assessed whether maternal immunization affects the avidity of infant vaccine-induced IgG antibodies, in the frame of a prospective clinical trial on pregnancy vaccination in Belgium. Infants born from Tdap (Boostrix®) vaccinated (N = 55) and unvaccinated (N = 26) mothers were immunized with a hexavalent pertussis containing vaccine (Infanrix Hexa®) at 8, 12 and 16 weeks, followed by a fourth dose at 15 months of age. Right before and one month after this fourth vaccine dose, the avidity of IgG antibodies against diphtheria toxin (DT), tetanus toxin (TT), pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (Prn) was determined using 1.5 M ammonium thiocyanate as dissociating agent. In both groups, antibody avidity was moderate for TT, PT, FHA and Prn and low for DT after priming. After a fourth dose, antibody avidity increased significantly to high avidity for TT and PT, whereas it remained moderate for FHA and Prn and low for DT. The avidity correlated positively with antibody level in both study groups, yet not significantly for PT. When comparing both study groups, only PT-specific antibodies showed significantly lower avidity in infants born from vaccinated than from unvaccinated mothers after the fourth vaccine dose. The clinical significance of lower avidity of vaccine induced infant antibodies after maternal vaccination, if any, needs further investigation.

  10. Usefulness of Toxoplasma gondii recombinant antigens (GRA1, GRA7 and SAG1) in an immunoglobulin G avidity test for the serodiagnosis of toxoplasmosis

    DEFF Research Database (Denmark)

    Pietkiewicz, H; Hiszczyńska-Sawicka, E; Kur, J

    2007-01-01

    The precise diagnosis of an acute and recent Toxoplasma infection in pregnant women and the newborn child is important before treatment. This study describes a new Toxoplasma gondii IgG avidity test based on a combination of recombinant GRA1, GRA7 and SAG1 antigens and shows that this test is use...

  11. Becoming a Reader: Significant Social Influences on Avid Book Readers

    Science.gov (United States)

    Merga, Margaret K.

    2017-01-01

    Understanding how social influences can foster avid book reader identification is a key research goal that warrants further investigation beyond a limited early-years lens. The author's 2015 International Study of Avid Book Readers (ISABR) explored, as one of its key research questions, the influence positive social agents can have on avid book…

  12. Direct block scheduling technology: Analysis of Avidity

    Directory of Open Access Journals (Sweden)

    Felipe Ribeiro Souza

    Full Text Available Abstract This study is focused on Direct Block Scheduling testing (Direct Multi-Period Scheduling methodology which schedules mine production considering the correct discount factor of each mining block, resulting in the final pit. Each block is analyzed individually in order to define the best target period. This methodology presents an improvement of the classical methodology derived from Lerchs-Grossmann's initial proposition improved by Whittle. This paper presents the differences between these methodologies, specially focused on the algorithms' avidity. Avidity is classically defined by the voracious search algorithms, whereupon some of the most famous greedy algorithms are Branch and Bound, Brutal Force and Randomized. Strategies based on heuristics can accentuate the voracity of the optimizer system. The applied algorithm use simulated annealing combined with Tabu Search. The most avid algorithm can select the most profitable blocks in early periods, leading to higher present value in the first periods of mine operation. The application of discount factors to blocks on the Lerchs-Grossmann's final pit has an accentuated effect with time, and this effect may make blocks scheduled for the end of the mine life unfeasible, representing a trend to a decrease in reported reserves.

  13. Molecular cloning and differential IgG responses to a histidine-rich ...

    African Journals Online (AJOL)

    C1 immunoglobulin G (IgG) subclass levels were assessed by ELISA in 15 pairs and 18 pairs of selected and cross-matched infected and putatively immune subjects from Cameroon and Ecuador, respectively. IgG3 and IgG4 levels were shown to be significantly higher in putatively immune (immune protected) subjects.

  14. Are Avid Readers Lurking in Your Language Arts Classroom? Myths of the Avid Adolescent Reader

    Science.gov (United States)

    Wilson, Nance S.; Kelley, Michelle J.

    2010-01-01

    This article describes a pilot study conducted with 10 identified avid adolescent readers who completed the Adolescent Motivation to Read Profile (AMRP) (Pitcher, Albright, DeLaney, Walker, Seunarienesingh, & Moggie, 2007) that includes both a survey to determine students' self-concept and value of reading and an interview that sheds light on what…

  15. Evaluation of cysticercus-specific IgG (total and subclasses and IgE antibody responses in cerebrospinal fluid samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies

    Directory of Open Access Journals (Sweden)

    Lisandra Akemi Suzuki

    Full Text Available In the present study, an enzyme-linked immunosorbent assay (ELISA standardized with vesicular fluid of Taenia solium cysticerci was used to screen for IgG (total and subclasses and IgE antibodies in cerebrospinal fluid (CSF samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies and patients with other neurological disorders. The following results were obtained: IgG-ELISA: 100% sensitivity (median of the ELISA absorbances (MEA=1.17 and 100% specificity; IgG1-ELISA: 72.7% sensitivity (MEA=0.49 and 100% specificity; IgG2-ELISA: 81.8% sensitivity (MEA=0.46 and 100% specificity; IgG3-ELISA: 63.6% sensitivity (MEA=0.12 and 100% specificity; IgG4-ELISA: 90.9% sensitivity (MEA=0.85 and 100% specificity; IgE-ELISA 93.8% sensitivity (MEA=0.60 and 100% specificity. There were no significant differences between the sensitivities and specificities in the detection of IgG-ELISA and IgE-ELISA, although in CSF samples from patients with neurocysticercosis the MEA of the IgG-ELISA was significantly higher than that of the IgE-ELISA. The sensitivity and MEA values of the IgG4-ELISA were higher than the corresponding values for the other IgG subclasses. Future studies should address the contribution of IgG4 and IgE antibodies to the physiopathology of neurocysticercosis.

  16. Kinetic and HPV infection effects on cross-type neutralizing antibody and avidity responses induced by Cervarix®

    Science.gov (United States)

    Kemp, Troy J.; Safaeian, Mahboobeh; Hildesheim, Allan; Pan, Yuanji; Penrose, Kerri J.; Porras, Carolina; Schiller, John T.; Lowy, Douglas R.; Herrero, Rolando; Pinto, Ligia A.

    2012-01-01

    Background We previously demonstrated that Cervarix® elicits antibody responses against vaccine-related types for which clinical efficacy was demonstrated (HPV-31 and -45). Here, we evaluated the kinetics of neutralization titers and avidity of Cervarix®-induced antibodies up to 36 months of follow-up in unexposed and HPV infected women. Methods A subset of women who participated in the Cost Rica HPV-16/18 Vaccine Trial had pre- and post-vaccination sera tested for antibody responses to HPV-16, -18, -31, -45, and -58 using a pseudovirion-based neutralization assay, and HPV-16 antibody avidity using an HPV-16 L1 VLP (virus-like particle)-based ELISA developed in our laboratory. Results In uninfected women, neutralizing antibody titers did not reach significance until after the 3rd dose for HPV-31 (month 12, p=0.009) and HPV-45 (month 12, p=0.003), but then persisted up to month 36 (HPV-31, p=0.01; HPV-45, p=0.002). Individuals infected with HPV-16 or HPV-31 at enrollment developed a significantly higher median antibody response to the corresponding HPV type after one dose, but there was not a difference between median titers after three doses compared to the HPV negative group. Median HPV-16 antibody avidity and titer increased over time up to month 12; however, the HPV-16 avidity did not correlate well with HPV-16 neutralizing antibody titers at each time point examined, except for month 6. The median avidity levels were higher in HPV-16 infected women at month 1 (p=0.04) and lower in HPV-16 infected women at month 12 (p=0.006) compared to the HPV negative women. Conclusions The persistence of cross-neutralization titers at month 36 suggests cross-reactive antibody responses are likely to persist long-term and are not influenced by infection status at enrollment. However, the weak correlation between avidity and neutralization titers emphasizes the need for examining avidity in efficacy studies to determine if high avidity antibodies play a critical role in

  17. AVID Teacher Leadership: Administrator and Teacher Perceptions of Leadership Attributes

    Science.gov (United States)

    Mills, Shirley J.; Huerta, Jeffery J.; Watt, Karen M.; Martinez, Jorge

    2014-01-01

    This study examined the perceptions of teachers and administrators with regard to AVID (Advancement via Individual Determination) teacher leaders. The purpose was to compare whether teachers and administrators agree on the types of attributes needed for teacher leaders involved in implementing AVID as a school reform effort. Results revealed that…

  18. Maternal immunoglobulin G avidity as a diagnostic tool to identify pregnant women at risk of congenital cytomegalovirus infection.

    Science.gov (United States)

    Kaneko, Masatoki; Ohhashi, Masanao; Minematsu, Toshio; Muraoka, Junsuke; Kusumoto, Kazumi; Sameshima, Hiroshi

    2017-03-01

    The immunoglobulin (Ig) G avidity index (AI) is useful to detect primary cytomegalovirus (CMV) infection. However, because IgG matures with time, this index is not useful to detect a primary infection, unless measured at an appropriate time. We aimed to clarify the difference between using IgG AI and IgM positivity according to the stage of pregnancy to identify congenital CMV infection risk. We collected the serum samples from 1115 pregnant women who underwent maternal screening for primary infection (n = 956) and were referred to our hospital because of CMV IgM positivity (n = 155) or had abnormal fetal ultrasonography findings (n = 4). The same sera samples were used to measure CMV IgM, IgG, and IgG AI. An IgG AI of congenital infection. Fourteen mothers gave birth to infected neonates. The sensitivity, specificity, and negative predictive values of the low IgG AI group with IgM-positive samples to discriminate between women with congenital infection at ≤14 weeks of gestation were 83.3, 83.8, and 99.1, respectively, which were higher than those of other subjects. Uni- and multivariate analyses revealed that IgM positivity and low IgG AI were independent variables associated with congenital infection at any stage of pregnancy, except low IgG AI at ≥15 weeks of gestation. Low IgG AI with IgM positivity at ≤14 weeks of gestation was a good indicator of congenital infection, which should prove useful in obstetric practice. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  19. ELISA technique standardisation for human toxocariasis diagnosis

    International Nuclear Information System (INIS)

    Espinoza, Y.; Suarez, R.; Huiza, A.

    2003-01-01

    To standardise ELISA technique for toxocara canis human infection diagnosis by using excreted-secreted antigen prepared in our country. T. canis eggs were collected by incubation with formalin (2%) at 28 o C in order to obtain third stage larvae that were freed and incubated in RPMI at 37 o C for 7 days; the medium was replaced by a similar one and stored at -20 o C. Antigen was concentrated and protein dosage was made. Sera from patients with toxocariasis and newborns were used as positive and negative controls by ELISA technique, dilutions 1/4 to 1/1024. Polystyrene plates were sensitised with antigen in several concentrations and conjugated peroxidase with horseradish IgG, anti human IgG and substrate OPD were used. Absorbance was read with spectrophotometer (Multiskan plus labsystems) at 492 nm. Cut off point was determined by negative sera absorbencies arithmetic mean plus 3 standard deviations. Antigen concentration was 50 ug/mL, sera dilution 1/128, conjugate dilution 1/1000 with optical density above 0,241. ELISA technique for serologic diagnosis of human infection by toxocara canis could be used in epidemiological studies in our country. Its efficacy will be determined in future studies

  20. Inflammatory pseudotumor: A gallium-avid mobile mesenteric mass

    International Nuclear Information System (INIS)

    Auringer, S.T.; Scott, M.D.; Sumner, T.E.

    1991-01-01

    An 8-yr-old boy with a 1-mo history of culture-negative fever and anemia underwent gallium, ultrasound, and computed tomography studies as part of the evaluation of a fever of unknown origin. These studies revealed a mobile gallium-avid solid abdominal mass subsequently proven to be an inflammatory pseudotumor of the mesentery, a rare benign mass. This report documents the gallium-avid nature of this rare lesion and discusses associated characteristic clinical, pathologic, and radiographic features

  1. Anti-dog IgG secondary antibody successfully detects IgG in a variety of aquatic mammals

    Science.gov (United States)

    Roehl, Katherine; Jankowski, Mark D.; Hofmeister, Erik K.

    2016-01-01

    Serological tests play an important role in the detection of wildlife diseases. However, while there are many commercial assays and reagents available for domestic species, there is a need to develop efficient serological assays for wildlife. In recent years, marine mammals have represented a wildlife group with emerging infectious diseases, such as influenza, brucellosis, and leptospirosis. However, with the exception of disease-agent-specific assays or functional assays, few reports describe the use of antibody detection assays in marine mammals. In an indirect enzyme-linked immunoassay (EIA) or an immunofluorescence assay, antibody is detected using an antitarget species secondary conjugated antibody. The sensitivity of the assay depends on the avidity of the binding reaction between the bound antibody and the detection antibody. A commercial polyclonal antidog IgG conjugated antibody was tested in an EIA for its ability to sensitively detect the IgG of seven marine mammals including sea otter (Enhydra lutris), polar bear (Ursus maritimus), grey seal (Halichoerus grypus), harbor seal (Phoca vitulina), northern elephant seal (Mirounga angustirostris), California sea lion (Zalophus californianus), Pacific walrus (Odobenus rosmarus) and one freshwater mammal: Asian small-clawed otter (Aonyx cinerea). With the exception of Asian small-clawed sea otters, the detection of IgG in these marine mammals either exceeded or was nearly equal to detection of dog IgG. The use of the tested commercial antidog IgG antibody may be a valid approach to the detection of antibody response to disease in sea mammals.

  2. The Dot Blot ELISA.

    Science.gov (United States)

    Gerbig, Donald G., Jr.; Fenk, Christopher J.; Goodhart, Amy S.

    2000-01-01

    Uses two laboratory techniques, Enzyme Linked Immunosorbent Assay (ELISA) and Western Blot, to demonstrate antibody-antigen binding concepts. Includes a list of required materials and directions for the procedure, and makes suggestions for classroom applications. (Contains 13 references.) (YDS)

  3. Comparison of dot-ELISA and standard ELISA for detection of Neisseria meningitidis outer membrane complex-specific antibodies

    Directory of Open Access Journals (Sweden)

    Elza FT Belo

    Full Text Available Dot-ELISA using the outer membrane complex antigens of Neisseria meningitidis as a target was standardized for rapid detection of meningococcal-specific antibodies in human serum. We investigated the level of meningococcal-specific IgG, IgA, and IgM in serum using dot-ELISA with outer membrane antigens prepared from Neisseria meningitidis serotype B:4.19:P1.15,3,7,9 (a strain isolated from a Brazilian epidemic. The dot-ELISA is based on the same principles as the standard ELISA and is useful for detection of anti-N. meningitidis B antibodies in serum of patients with meningococcal infections. For the assay, outer membrane complexes (OMCs were absorbed by nitrocellulose membrane and blocked with a 5% skim milk solution. Serum samples were drawn upon hospital admission and during convalescence from patients with meningococcal septicemia, and single samples were drawn from uninfected controls. We retrospectively examined a total of 57 serum samples: 35 from patients infected with N. meningitidis B, 12 from patients infected with Haemophilus influenzae b, and 10 from health individuals. When performed at room temperature, dot-ELISA took approximately four hours to perform, and the optimum antigen concentration was 0.42 µg per dot. The specificity of IgG, IgM, and IgA demonstrates that dot-ELISA using OMCs from N. meningitidis B as a target is suitable for serologic verification of clinically suspected meningococcal disease in patients and for titer determination of antibodies produced during different phases of natural infection. Furthermore, the sensitivity of dot-ELISA was comparable to that of standard ELISA. Overall, dot-ELISA is simple to perform, rapid, and low cost. Further validation of the test as a screening tool is required.

  4. Cornelius Elisa Bertus Bremekamp

    NARCIS (Netherlands)

    Lanjouw, J.

    1969-01-01

    Cornelis Elisa Bertus Bremekamp was born at Dordrecht on February 7th 1888. He is therefore now just past eighty, and he has been a member of the Koninklijke Nederlandse Botanische Vereniging for sixty years. He studied at the Utrecht State University and, like many of his contemporaries, was

  5. A quantitative comparative analysis of Advancement via Independent Determination (AVID) in Texas middle schools

    Science.gov (United States)

    Reed, Krystal Astra

    The "Advancement via Individual Determination (AVID) program was designed to provide resources and strategies that enable underrepresented minority students to attend 4-year colleges" (AVID Center, 2013, p. 2). These students are characterized as the forgotten middle in that they have high test scores, average-to-low grades, minority or low socioeconomic status, and will be first-generation college students (AVID, 2011). Research indicates (Huerta, Watt, & Butcher, 2013) that strict adherence to 11 program components supports success of students enrolled in AVID, and AVID certification depends on districts following those components. Several studies (AVID Center, 2013) have investigated claims about the AVID program through qualitative analyses; however, very few have addressed this program quantitatively. This researcher sought to determine whether differences existed between student achievement and attendance rates between AVID and non-AVID middle schools. To achieve this goal, the researcher compared eighth-grade science and seventh- and eighth-grade mathematics scores from the 2007 to 2011 Texas Assessment of Knowledge and Skills (TAKS) and overall attendance rates in demographically equivalent AVID and non-AVID middle schools. Academic Excellence Indicator System (AEIS) reports from the Texas Education Agency (TEA) were used to obtain 2007 to 2011 TAKS results and attendance information for the selected schools. The results indicated a statistically significant difference between AVID demonstration students and non-AVID students in schools with similar CI. No statistically significant differences were found on any component of the TAKS for AVID economically disadvantaged students. The mean scores indicated an achievement gap between non-AVID and AVID demonstration middle schools. The findings from the other three research questions indicated no statistically significant differences between AVID and non-AVID student passing rates on the seventh- and eighth

  6. Adenovirus-specific IgG maturation as a surrogate marker in acute exacerbations of COPD.

    Science.gov (United States)

    Boeck, Lucas; Gencay, Mikael; Roth, Michael; Hirsch, Hans H; Christ-Crain, Mirjam; Mueller, Beat; Tamm, Michael; Stolz, Daiana

    2014-08-01

    B cells in airways and lung parenchyma may be involved in COPD evolution; however, whether their pathogenic role is beneficial or harmful remains controversial. The objective of this study was to investigate the maturation of adenovirus-specific immunoglobulins in patients with COPD with respect to clinical outcome. The presence of adenovirus-specific immunoglobulins during acute exacerbation of COPD (AECOPD) was analyzed at exacerbation and 2 to 3 weeks later. Patients with detectable adenovirus-specific IgM and low IgG avidity were grouped into fast and delayed IgG maturation. The clinical outcome of both groups was evaluated. Of 208 patients, 43 (20.7%) had serologic evidence of recent adenovirus infection and were grouped by fast IgG maturation (26 patients) and delayed IgG maturation (17 patients). Baseline characteristics, AECOPD therapy, and duration of hospitalization were similar in both groups, but the AECOPD recurrence rate within 6 months was higher (P = .003), and there was a trend for earlier AECOPD-related rehospitalizations (P = .061) in the delayed IgG maturation group. The time to rehospitalization or death within 2 years was shorter in patients with delayed IgG maturation (P = .003). Adenovirus-specific IgG maturation was an independent predictor of the number of AECOPD recurrences within 6 months (P = .001) and the occurrence of hospitalization or death within 2 years (P = .005). Delayed immunoglobulin avidity maturation following COPD exacerbation is associated with worse outcomes. ISRCTN Register; No.: ISRCTN77261143; URL: www.isrctn.org.

  7. The Role of Leaders in AVID Schools and the Impact on Student Achievement

    OpenAIRE

    Noone, Colleen Callahan

    2010-01-01

    The Advancement Via Individual Determination (AVID) program is designed to provide resources and strategies to enable underrepresented minority students to attend four-year colleges. Research indicates that the success of the students enrolled in AVID depends on the strict adherence to the AVID components. The purpose of this study was to determine whether the principal and administratorâ s understanding of, commitment to and involvement in AVID accounted for the difference in student achi...

  8. Anti-food and anti-microbial IgG subclass antibodies in inflammatory bowel disease.

    Science.gov (United States)

    Jansen, Anke; Mandić, Ana D; Bennek, Eveline; Frehn, Lisa; Verdier, Julien; Tebrügge, Irene; Lutz, Holger; Streetz, Konrad; Trautwein, Christian; Sellge, Gernot

    2016-12-01

    Inflammatory bowel disease (IBD), particularly Crohn's disease (CD), is associated with increased microbial-specific IgG and IgA antibodies, whereas alterations of anti-food antibodies are still disputed. The knowledge about IgG subclass antibodies in IBD is limited. In this study we analysed IgG subclass antibodies specific for nutritional and commensal antigens in IBD patients and controls. Serum IgG1, IgG2, IgG3 and IgG4 specific for wheat and milk extracts, purified ovalbumin, Escherichia coli and Bacteroides fragilis lysates and mannan from Saccharomyces cerevisiae were analysed by ELISA in patients with CD (n = 56), ulcerative colitis (UC; n = 29), acute gastroenteritis/colitis (n = 12) as well as non-inflammatory controls (n = 62). Anti-Saccharomyces cerevisiae antibodies (ASCA) of all IgG subclasses and anti-B. fragilis IgG1 levels were increased in CD patients compared to UC patients and controls. The discriminant validity of ASCA IgG2 and IgG4 was comparable with that of ASCA pan-IgG and IgA, whereas it was inferior for ASCA IgG1/IgG3 and anti-B. fragilis IgG1. Complicated CD defined by the presence of perianal, stricturing or penetrating disease phenotypes was associated with increased ASCA IgG1/IgG3/IgG4, anti-B. fragilis IgG1 and anti-E. coli IgG1 levels. Anti-food IgG subclass levels were not different between IBD patients and controls and did not correlate with food intolerance. In contrast to anti-microbial Abs, food-specific IgG responses were predominately of the IgG4 isotype and all food-specific IgG subclass levels correlated negatively with age. Our study supports the notion that the adaptive immune recognition of food and commensal antigens are differentially regulated.

  9. Evaluation of cysticercus-specific IgG (total and subclasses and IgE antibody responses in cerebrospinal fluid samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies Avaliação das respostas de anticorpos anti-cisticercos IgG (total e subclasses e IgE em amostras de líquido cefalorraquidiano de pacientes com neurocisticercose apresentando produção intratecal de anticorpos específicos IgG

    Directory of Open Access Journals (Sweden)

    Lisandra Akemi Suzuki

    2013-01-01

    Full Text Available In the present study, an enzyme-linked immunosorbent assay (ELISA standardized with vesicular fluid of Taenia solium cysticerci was used to screen for IgG (total and subclasses and IgE antibodies in cerebrospinal fluid (CSF samples from patients with neurocysticercosis showing intrathecal production of specific IgG antibodies and patients with other neurological disorders. The following results were obtained: IgG-ELISA: 100% sensitivity (median of the ELISA absorbances (MEA=1.17 and 100% specificity; IgG1-ELISA: 72.7% sensitivity (MEA=0.49 and 100% specificity; IgG2-ELISA: 81.8% sensitivity (MEA=0.46 and 100% specificity; IgG3-ELISA: 63.6% sensitivity (MEA=0.12 and 100% specificity; IgG4-ELISA: 90.9% sensitivity (MEA=0.85 and 100% specificity; IgE-ELISA 93.8% sensitivity (MEA=0.60 and 100% specificity. There were no significant differences between the sensitivities and specificities in the detection of IgG-ELISA and IgE-ELISA, although in CSF samples from patients with neurocysticercosis the MEA of the IgG-ELISA was significantly higher than that of the IgE-ELISA. The sensitivity and MEA values of the IgG4-ELISA were higher than the corresponding values for the other IgG subclasses. Future studies should address the contribution of IgG4 and IgE antibodies to the physiopathology of neurocysticercosis.No presente estudo, uma reação imunoenzimática (ELISA padronizada com o fluido vesicular de cisticercos de Taenia solium foi utilizada para avaliar as respostas de anticorpos anti-cisticercos IgG (total e subclasses e IgE em amostras de líquido cefalorraquidiano (LCR de pacientes com neurocisticercose apresentando produção intratecal de anticorpos específicos IgG e pacientes com outras desordens neurológicas. Os seguintes resultados foram obtidos: ELISA-IgG: 100% de sensibilidade (mediana das absorbâncias das reações ELISA (MAE=1,17 e especificidade 100%; ELISA-IgG1: sensibilidade 72,7% (MAE=0,49 e especificidade 100%; ELISA-IgG2

  10. Nasal Immunization Confers High Avidity Neutralizing Antibody Response and Immunity to Primary and Recurrent Genital Herpes in Guinea Pigs.

    Science.gov (United States)

    Persson, Josefine; Zhang, Yuan; Olafsdottir, Thorunn A; Thörn, Karolina; Cairns, Tina M; Wegmann, Frank; Sattentau, Quentin J; Eisenberg, Roselyn J; Cohen, Gary H; Harandi, Ali M

    2016-01-01

    Genital herpes is one of the most prevalent sexually transmitted infections in both the developing and developed world. Following infection, individuals experience life-long latency associated with sporadic ulcerative outbreaks. Despite many efforts, no vaccine has yet been licensed for human use. Herein, we demonstrated that nasal immunization with an adjuvanted HSV-2 gD envelope protein mounts significant protection to primary infection as well as the establishment of latency and recurrent genital herpes in guinea pigs. Nasal immunization was shown to elicit specific T cell proliferative and IFN-γ responses as well as systemic and vaginal gD-specific IgG antibody (Ab) responses. Furthermore, systemic IgG Abs displayed potent HSV-2 neutralizing properties and high avidity. By employing a competitive surface plasmon resonance (SPR) analysis combined with a battery of known gD-specific neutralizing monoclonal Abs (MAbs), we showed that nasal immunization generated IgG Abs directed to two major discontinuous neutralizing epitopes of gD. These results highlight the potential of nasal immunization with an adjuvanted HSV-2 envelope protein for induction of protective immunity to primary and recurrent genital herpes.

  11. Nasal Immunization Confers High Avidity Neutralizing Antibody Response and Immunity to Primary and Recurrent Genital Herpes in Guinea Pigs

    Science.gov (United States)

    Persson, Josefine; Zhang, Yuan; Olafsdottir, Thorunn A.; Thörn, Karolina; Cairns, Tina M.; Wegmann, Frank; Sattentau, Quentin J.; Eisenberg, Roselyn J.; Cohen, Gary H.; Harandi, Ali M.

    2016-01-01

    Genital herpes is one of the most prevalent sexually transmitted infections in both the developing and developed world. Following infection, individuals experience life-long latency associated with sporadic ulcerative outbreaks. Despite many efforts, no vaccine has yet been licensed for human use. Herein, we demonstrated that nasal immunization with an adjuvanted HSV-2 gD envelope protein mounts significant protection to primary infection as well as the establishment of latency and recurrent genital herpes in guinea pigs. Nasal immunization was shown to elicit specific T cell proliferative and IFN-γ responses as well as systemic and vaginal gD-specific IgG antibody (Ab) responses. Furthermore, systemic IgG Abs displayed potent HSV-2 neutralizing properties and high avidity. By employing a competitive surface plasmon resonance (SPR) analysis combined with a battery of known gD-specific neutralizing monoclonal Abs (MAbs), we showed that nasal immunization generated IgG Abs directed to two major discontinuous neutralizing epitopes of gD. These results highlight the potential of nasal immunization with an adjuvanted HSV-2 envelope protein for induction of protective immunity to primary and recurrent genital herpes. PMID:28082979

  12. Increased levels of IgG antibodies against human HSP60 in patients with spondyloarthritis

    DEFF Research Database (Denmark)

    Hjelholt, Astrid; Carlsen, Thomas; Deleuran, Bent

    2013-01-01

    severity in relation to HLA-B27 was evaluated.Serum samples from 82 patients and 50 controls were analysed by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (Ig)G1, IgG2, IgG3 and IgG4 antibodies against human HSP60 and HSP60 from Chlamydia trachomatis, Salmonella enteritidis...... bacterial and human HSP60, also named HSPD1, are highly homologous, cross-reactivity has been suggested in disease initiation. In this study, levels of antibodies against bacterial and human HSP60 were analysed in SpA patients and healthy controls, and the association between such antibodies and disease...... and Campylobacter jejuni. Disease severity was assessed by the clinical scorings Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI) and Bath Ankylosing Spondylitis Metrology Index (BASMI). Levels of IgG1 and IgG3 antibodies against human HSP60...

  13. AVID: An integrative framework for discovering functional relationships among proteins

    Directory of Open Access Journals (Sweden)

    Keating Amy E

    2005-06-01

    Full Text Available Abstract Background Determining the functions of uncharacterized proteins is one of the most pressing problems in the post-genomic era. Large scale protein-protein interaction assays, global mRNA expression analyses and systematic protein localization studies provide experimental information that can be used for this purpose. The data from such experiments contain many false positives and false negatives, but can be processed using computational methods to provide reliable information about protein-protein relationships and protein function. An outstanding and important goal is to predict detailed functional annotation for all uncharacterized proteins that is reliable enough to effectively guide experiments. Results We present AVID, a computational method that uses a multi-stage learning framework to integrate experimental results with sequence information, generating networks reflecting functional similarities among proteins. We illustrate use of the networks by making predictions of detailed Gene Ontology (GO annotations in three categories: molecular function, biological process, and cellular component. Applied to the yeast Saccharomyces cerevisiae, AVID provides 37,451 pair-wise functional linkages between 4,191 proteins. These relationships are ~65–78% accurate, as assessed by cross-validation testing. Assignments of highly detailed functional descriptors to proteins, based on the networks, are estimated to be ~67% accurate for GO categories describing molecular function and cellular component and ~52% accurate for terms describing biological process. The predictions cover 1,490 proteins with no previous annotation in GO and also assign more detailed functions to many proteins annotated only with less descriptive terms. Predictions made by AVID are largely distinct from those made by other methods. Out of 37,451 predicted pair-wise relationships, the greatest number shared in common with another method is 3,413. Conclusion AVID provides

  14. Development of a sandwich ELISA for quantification of immunoglobulin G in mink blood

    DEFF Research Database (Denmark)

    Mathiesen, Ronja; Chriél, Mariann; Struve, T.

    2016-01-01

    early immunity and thus their resistance against pathogenic agents found in the environment. This study describes a sandwich ELISA for quantification of the concentration of total immunoglobulin G in mink blood. The ELISA was validated with serum samples from females (n=8) and their kits (litters of 4......-12). Preliminary results show that the IgG concentration among kits from the same litter was similar, while litter to litter variation was high....

  15. Kinetics of IgG antibody to cytomegalovirus (CMV after birth and seroprevalence of anti-CMV IgG in Chinese children

    Directory of Open Access Journals (Sweden)

    Chen Jie

    2012-12-01

    Full Text Available Abstract Background Prevalence of cytomegalovirus (CMV infection is 90–100% in developing countries; however, the kinetics of anti-CMV IgG in infants remains elusive. Methods Sera from 112 mother-newborn pairs and longitudinal samples from 41 infants up to 2-year old were tested for anti-CMV IgG and IgM. Additionally, samples from 837 healthy children were included. Results Of 112 mothers, 108 (96.4% were anti-CMV IgG positive; their 108 newborns were also seropositive. In a 2-year follow-up among 40 infants of positive mothers, anti-CMV IgG level in 8 individuals decreased with time and became undetectable by age of 3.5–8 months, and that in 32 others decreased at 1- and 3.5-month old, and then increased. Based on the positive IgM, rising IgG levels, and low anti-CMV IgG avidity index, 76.7% of the primary infections were demonstrated to occur during 1–3.5 months of age. The overall seroprevalence of anti-CMV in 837 children was 82.4%, which was generally constant from 2 to 8 years old (χ2 = 3.150, p = 0.790. Conclusions The maternally acquired anti-CMV IgG in infants disappears before 8-month old. Primary CMV infection in Chinese children mostly occurs during 1–3.5 months of age. Whether the relatively lower seroprevalence of anti-CMV in Chinese children found in this survey may reflect the positive rate in child-bearing age women in the future remains to be further studied.

  16. Kinetics of IgG antibody to cytomegalovirus (CMV) after birth and seroprevalence of anti-CMV IgG in Chinese children.

    Science.gov (United States)

    Chen, Jie; Hu, Lingqing; Wu, Meiling; Zhong, Tianying; Zhou, Yi-Hua; Hu, Yali

    2012-12-10

    Prevalence of cytomegalovirus (CMV) infection is 90-100% in developing countries; however, the kinetics of anti-CMV IgG in infants remains elusive. Sera from 112 mother-newborn pairs and longitudinal samples from 41 infants up to 2-year old were tested for anti-CMV IgG and IgM. Additionally, samples from 837 healthy children were included. Of 112 mothers, 108 (96.4%) were anti-CMV IgG positive; their 108 newborns were also seropositive. In a 2-year follow-up among 40 infants of positive mothers, anti-CMV IgG level in 8 individuals decreased with time and became undetectable by age of 3.5-8 months, and that in 32 others decreased at 1- and 3.5-month old, and then increased. Based on the positive IgM, rising IgG levels, and low anti-CMV IgG avidity index, 76.7% of the primary infections were demonstrated to occur during 1-3.5 months of age. The overall seroprevalence of anti-CMV in 837 children was 82.4%, which was generally constant from 2 to 8 years old (χ2 = 3.150, p = 0.790). The maternally acquired anti-CMV IgG in infants disappears before 8-month old. Primary CMV infection in Chinese children mostly occurs during 1-3.5 months of age. Whether the relatively lower seroprevalence of anti-CMV in Chinese children found in this survey may reflect the positive rate in child-bearing age women in the future remains to be further studied.

  17. Seroprevalence of Toxoplasma gondii in wild kangaroos using an ELISA.

    Science.gov (United States)

    Parameswaran, N; O'Handley, R M; Grigg, M E; Fenwick, S G; Thompson, R C A

    2009-06-01

    Infection with Toxoplasma gondii is a significant problem in Australian marsupials, and can lead to devastating disease and predispose animals to predation. T. gondii infection in kangaroos is also of public health significance due to the kangaroo meat trade. A moderate seroprevalence of T. gondii was observed in a study of western grey kangaroos located in the Perth metropolitan area in Western Australia. Of 219 kangaroos tested, 15.5% (95%CI: 10.7-20.3) were positive for T. gondii antibodies using an ELISA developed to detect T. gondii IgG in macropod marsupials. When compared with the commercially available MAT (modified agglutination test), the ELISA developed was in absolute agreement and yielded a kappa coefficient of 1.00. Of 18 kangaroos tested for the presence of T. gondii DNA by PCR, the 9 ELISA positive kangaroos tested PCR positive and the 9 ELISA negative kangaroos tested PCR negative indicating the ELISA protocol was both highly specific and sensitive and correlated 100% with the more labour intensive PCR assay.

  18. A new ELISA for determination of potency in snake antivenoms.

    Science.gov (United States)

    Rial, A; Morais, V; Rossi, S; Massaldi, H

    2006-09-15

    A competitive ELISA for potency determination of bothropic equine antivenom was developed and compared to the conventional in vivo ED(50) assay, with the aim of partially substituting the in vivo assay in the monitoring of antivenom immunoglobulin levels. On this purpose, blood samples were taken at different times during and after the immunization protocol of the lot of horses used for production of snake antivenom at the Instituto de Higiene, Uruguay. Both the competitive ELISA and the ED(50) assay were performed on those samples. In addition, a group of five commercial pepsin-digested antivenoms were tested by both methods. A significant (P<0.001) correlation (Pearson's r=0.957) was found between the ELISA titres and the corresponding ED(50) values, indicating that the in vitro test can estimate the neutralizing antibody capacity of the sera as well as the in vivo assay. By means of this new ELISA, it was found that the immunized animals maintained good venom antibody titres, in the order of 20-50% of the maximum achieved, even 10 month after the end of the immunization schedule. The main advantage of our ELISA design is its ability to correctly estimate the neutralization capacity of crude hyperimmune plasma and antivenom sera independently of their antibody composition in terms of whole IgG or F(ab')(2) fragment.

  19. Elisa Biagini (Florencia, 1970

    Directory of Open Access Journals (Sweden)

    Berta González Saavedra

    2017-01-01

    Full Text Available Elisa Biagini vive en Italia tras haber estudiado y enseñado en los Estados Unidos durante varios años. Sus poesías han sido publicadas en varias revistas y antologías italianas y americanas, entre otras. Algunas de las más recientes son Nuovissima poesia italiana (Mondadori, 2004 y Parola plurale (Sosselam 2005. Ha publicado siete colecciones poéticas, algunas bilingües, entre las que figuran  L'Ospite (Einaudi, 2004, Fiato. Parole per musica (Edizionidif, 2006, Nel bosco (Einaudi, 2007, The guest in the wood (Chelsea editions, 2013 – 2014 Best Translated Book Award, y la reciente Da una crepa (Einaudi, 2014. Sus poesías han sido traducidas al inglés, español, francés, portugués, japonés, croata, eslovaco, alemán, albanés, ruso, árabe y chino. Ha participado en importantes festivales italianos e internacionales (entre otros, en Italia “Festival della Letteratura” de Mantua, “Festival Poesia” de Parma, “RomaPoesia” de Roma, y en el extranjero “Stanza- Scotland's International Poetry Festival” en St. Andrews, Escocia, “Dubai International Poetry Festival” en Emiratos Árabes Unidos, “PoesieFestivalBerlin” en Berlín, “International Writers Workshop” en Hong Kong, “Struga Poetry Festival” en Struga, Macedonia, “Poetry Parnassus” en Londres, “Printemps des poètes” en Luxemburgo, “Queensland Poetry Festival” en Brisbane, Australia, “Festival Internacional de Poesía de Granada” en Nicaragua, “Xu Zhimo Poetry and Art Festival” en el King's College de Cambridge. Asimismo es traductora de poesía americana y, además de editar algunas colecciones de poetisas americanas contemporáneas, se ha encargado de la edición del volumen Nuovi poeti americani (Einaudi, 2006. Infine, insegna Scrittura Creativa (poesia, Travel Writing e Storia dell'Arte in Italia e all’estero, además de colaborar con artistas visules, coreógrafos y músicos. Entre otras actividades, es artista visual. www.elisabiagini.it

  20. INVESTIGATION OF TOXOPLASMA INFECTIONS IN NATIVE POULTRY OF TABRIZ CITY USING ELISA AND INDIRECT IMMUNOFLUORESCENCE METHODS

    Directory of Open Access Journals (Sweden)

    Muhammad Fathollahzadeh

    2017-12-01

    Full Text Available Toxoplasma gondii is widely distributed in humans and other animals and birds throughout the world. Toxoplasmosis is a common zoonotic infection in the world and in immune-deficient patients, it may cause acute and lethal infection. The aim of current study was to investigate the prevalence of antibodies against Toxoplasma gondii IgG by ELISA and indirect immune fluorescence in backyard fowls in Tabriz city. Blood samples were evaluated to detect anti - Toxoplasma gondii IgG in backyard fowls. The prevalence of Antibody against Toxoplasma IgG in native poultry of Tabriz was detected 18%in ELISA and 30% in indirect immune-fluorescence (IFA method. Results of indirect IFA method indicated, 14% of roosters and 16% of hens were infected. Because of high rate of infection with Toxoplasma gondii in rural poultry, monitoring program along with anti-parasite treatment should be implemented.

  1. T cell avidity and tumor recognition: implications and therapeutic strategies

    Directory of Open Access Journals (Sweden)

    Roszkowski Jeffrey J

    2005-09-01

    Full Text Available Abstract In the last two decades, great advances have been made studying the immune response to human tumors. The identification of protein antigens from cancer cells and better techniques for eliciting antigen specific T cell responses in vitro and in vivo have led to improved understanding of tumor recognition by T cells. Yet, much remains to be learned about the intricate details of T cell – tumor cell interactions. Though the strength of interaction between T cell and target is thought to be a key factor influencing the T cell response, investigations of T cell avidity, T cell receptor (TCR affinity for peptide-MHC complex, and the recognition of peptide on antigen presenting targets or tumor cells reveal complex relationships. Coincident with these investigations, therapeutic strategies have been developed to enhance tumor recognition using antigens with altered peptide structures and T cells modified by the introduction of new antigen binding receptor molecules. The profound effects of these strategies on T cell – tumor interactions and the clinical implications of these effects are of interest to both scientists and clinicians. In recent years, the focus of much of our work has been the avidity and effector characteristics of tumor reactive T cells. Here we review concepts and current results in the field, and the implications of therapeutic strategies using altered antigens and altered effector T cells.

  2. Anti Helicobacter pylori IgG and IgA response in patients with gastric cancer and chronic gastritis.

    Science.gov (United States)

    Manojlovic, Nebojsa; Babic, Dragana; Filipovic-Ljeshovic, Ivana; Pilcevic, Dijana

    2008-01-01

    Immune response against Helicobacter pylori is important for the course and outcome of infection. We conducted study looking for the difference in anti H. pylori IgG and IgA between patients with intestinal type of gastric cancer, superficial and atrophic gastritis. For this study, 133 patients infected with H. pylori were enrolled: 50 with superficial gastritis, 42 with atrophic gastritis and 41 with gastric cancer. Anti H. pylori IgG and IgA ELISA tests were performed. The difference in antibody titers of IgG and IgA, frequency of IgA > IgG ratio and combination of low IgG and IgA > IgG ratio were analyzed. The patients with gastritis had higher titer of IgG that the patients with gastric cancer (p gastritis had higher titer of IgA than the patients with gastric cancer (p IgG ratio is more frequent in patients with gastric cancer than in the patients with superficial gastritis (p IgG is more frequent in the patients with gastric cancer than in the patients with gastritis (p cancer elicit different anti H. pylori IgG and IgA response than the patients with superficial and atrophic gastritis. Low IgG and IgA predominance seems characteristic for gastric cancer.

  3. Determination of rubella IgG and IgM antibodies among women of ...

    African Journals Online (AJOL)

    Rubella is an important Toga virus disease, causing significant human public health problem in Nigeria. In this study effort was made to determine rubella prevalence in women in Kano State. One thousand blood sera were tested with IgM specific Enzyme Immuno Assay (EIA) and IgG indirect solid phase ELISA based kits.

  4. Measurement of allergen-specific IgG in serum is of limited value for the management of dogs diagnosed with cutaneous adverse food reactions

    NARCIS (Netherlands)

    Hagen-Plantinga, E A; Leistra, M H G; Sinke, J D; Vroom, M W; Savelkoul, H F J; Hendriks, W H

    Conflicting results have been reported in the literature in terms of the usefulness of serological testing for IgG against food allergens in dogs with cutaneous adverse food reaction (CAFR). The aim of the present study was to evaluate the suitability of a commercially available IgG ELISA for

  5. Measurement of allergen-specific IgG in serum is of limited value for the management of dogs diagnosed with cutaneous adverse food reactions

    NARCIS (Netherlands)

    Hagen-Plantinga, E.A.; Leistra, M.H.G.; Sinke, J.D.; Vroom, M.W.; Savelkoul, H.F.J.; Hendriks, W.H.

    2017-01-01

    Conflicting results have been reported in the literature in terms of the usefulness of serological testing for IgG against food allergens in dogs with cutaneous adverse food reaction (CAFR). The aim of the present study was to evaluate the suitability of a commercially available IgG ELISA for

  6. Production and Purification of a Polyclonal Antibody Against Purified Mouse IgG2b in Rabbits Towards Designing Mouse Monoclonal Isotyping Kits

    Directory of Open Access Journals (Sweden)

    Sadeq Eivazi

    2015-03-01

    Full Text Available Purpose: Mouse IgG subclasses containing IgG1, IgG2a, IgG2b and IgG3 have been defined and described both physiochemically and immunologically. Methods: Sepharose beads conjugated with protein A affinity chromatography was used for purification of mouse IgG2b. Sodium citrate buffer (0.1 M, pH: 3.5 was used for separation of mouse IgG2b. Verification of the purified fractions was monitored by SDS-PAGE (polyacrylamide gel electrophoresis in reducing condition. Immunized rabbit serum was collected and precipitated at the final concentration of 50% ammonium sulfate. After dialysis against tris-phosphate buffer (pH: 8.1 ion exchange chromatography column was used for purification of rabbit anti-mouse IgG2b. The periodate method was performed for conjugation with some variations. After conjugation, direct ELISA was used to determine the titer of HRP conjugated rabbit IgG against mouse IgG2b. Results: The titer of rabbit anti-mouse IgG2b that determined by ELISA was 32000. The purity of rabbit anti-mouse IgG2b was about 95%. The optimum dilution of prepared HRP conjugated IgG was 1:10000. This study showed that ion-exchange chromatography and affinity chromatography could be appropriate techniques for purification of mouse IgG and IgG subclasses respectively. Conclusion: This study showed that affinity chromatography could be an appropriate method for purification of IgG2b antibodies.

  7. Pattern of pre-existing IgG subclass responses to a panel of asexual stage malaria antigens reported during the lengthy dry season in Daraweesh, Sudan

    DEFF Research Database (Denmark)

    Nasr, A; Iriemenam, N C; Troye-Blomberg, M

    2011-01-01

    The anti-malarial IgG immune response during the lengthy and dry season in areas of low malaria transmission as in Eastern Sudan is largely unknown. In this study, ELISA was used for the measurement of pre-existing total IgG and IgG subclasses to a panel of malaria antigens, MSP2-3D7, MSP2-FC27...

  8. Non-apical membrane antigen 1 (AMA1 IgGs from Malian children interfere with functional activity of AMA1 IgGs as judged by growth inhibition assay.

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    Kazutoyo Miura

    Full Text Available BACKGROUND: Apical membrane antigen 1 (AMA1 is one of the best-studied blood-stage malaria vaccine candidates. When an AMA1 vaccine was tested in a malaria naïve population, it induced functionally active antibodies judged by Growth Inhibition Assay (GIA. However, the same vaccine failed to induce higher growth-inhibitory activity in adults living in a malaria endemic area. Vaccination did induce functionally active antibodies in malaria-exposed children with less than 20% inhibition in GIA at baseline, but not in children with more than that level of baseline inhibition. METHODS: Total IgGs were purified from plasmas collected from the pediatric trial before and after immunization and pools of total IgGs were made. Another set of total IgGs was purified from U.S. adults immunized with AMA1 (US-total IgG. From these total IgGs, AMA1-specific and non-AMA1 IgGs were affinity purified and the functional activity of these IgGs was evaluated by GIA. Competition ELISA was performed with the U.S.-total IgG and non-AMA1 IgGs from malaria-exposed children. RESULTS: AMA1-specific IgGs from malaria-exposed children and U.S. vaccinees showed similar growth-inhibitory activity at the same concentrations. When mixed with U.S.-total IgG, non-AMA1 IgGs from children showed an interference effect in GIA. Interestingly, the interference effect was higher with non-AMA1 IgGs from higher titer pools. The non-AMA1 IgGs did not compete with anti-AMA1 antibody in U.S.-total IgG in the competition ELISA. CONCLUSION: Children living in a malaria endemic area have a fraction of IgGs that interferes with the biological activity of anti-AMA1 antibody as judged by GIA. While the mechanism of interference is not resolved in this study, these results suggest it is not caused by direct competition between non-AMA1 IgG and AMA1 protein. This study indicates that anti-malaria IgGs induced by natural exposure may interfere with the biological effect of antibody induced by an AMA1

  9. Antibody isotypes, including IgG subclasses, in Ecuadorian patients with pulmonary Paragonimiasis

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    Angel Guevara E.

    1995-08-01

    Full Text Available An ELISA test was developed to detect Paragonimus-specific antibodies, including IgG subclasses, using P. mexicanus crude water-soluble antigens. The test was standardized to detect antibodies in sera of Ecuadorian patients with pulmonary paragonimiasis and negative controls from the endemic area. The detected mean levels of IgG (0.753, SEM: 0.074 and IgM (0.303, SEM: 0.033 were significantly elevated (P<0.05. Within the IgG subclasses, IgG4 showed the highest detected mean level (0.365, SEM: 0.116 and the other three subclasses showed considerably lower mean levels (IgG1, 0.186 SEM: 0.06; IgG2, 0.046 SEM: 0.01; IgG3, 0.123 SEM: 0.047. The number of P. mexicanus eggs found in sputum of infected individuals showed a positive correlation with the level of antibodies detected for IgM, IgG and its subclasses (P<0.001. The relevance of these findings in Ecuadorian patients suffering from pulmonary paragonimiasis is discussed.

  10. Measurement of spesific IgG to 14 foods in the serum of 32 alleric patients

    International Nuclear Information System (INIS)

    Zou Hanbing; Xu Yiping

    2006-01-01

    To evaluate the serum specific IgG to 14 food allergens in allergic patients, the food specific IgG was measured by ELISA in 32 allergic patients as well as 22 normal subjects. Results showed that the food specific IgG increased in 32 allergic patients. The positive rates for allergic patients were: shrimp: 34.4%, peanut:21.9%, egg:18.8%, crab:15.6%, wheat:12. 5%, ling:9.4%, corn:6.3%, soja:6.3%, beef:3.1%, mushroom:3.1%, tomato:3.1%, chicken:0, pork:0, rice:0. Only low increased levels of specific IgG to egg and pork in normal subjects were found with the same positive rate of 4.5% and the specific IgG to other food were negative. The fact that the food specific IgG increased in allergic disease patients means that not only IgE but also IgG could be prodused in allergic patients and there exists some relation between the two antibodies. It suggests that the measurement of food specific IgG in patients suffering from food allergy might be useful for diagnosis, prevention and treatment for such patients. (authors)

  11. Predictive value of Borrelia burgdorferi IgG antibody levels in patients referred to a tertiary Lyme centre.

    Science.gov (United States)

    Zwerink, M; Zomer, T P; van Kooten, B; Blaauw, G; van Bemmel, T; van Hees, B C; Vermeeren, Y M; Landman, G W

    2017-12-24

    A two-step testing strategy is recommended in serological testing for Lyme borreliosis; positive and indeterminate enzyme-linked immunosorbent assay (ELISA) results are confirmed with immunoblots. Several ELISAs quantify the concentration of antibodies tested, however, no recommendation exists for an upper cut-off value at which an IgG ELISA is sufficient and the immunoblot can be omitted. The study objective was to determine at which IgG antibody level an immunoblot does not have any additional predictive value compared to ELISA results. Data of adult patients who visited a tertiary Lyme centre between 2008 and 2014 were analysed. Both an ELISA (Enzygnost Lyme link VlsE IgG) and immunoblot (recomLine blot Borrelia) were performed. Clinical data were extracted from the patient's digital medical record. Positive predictive values (PPVs) for either previous or active infection with Borrelia burgdorferi s.l. were calculated for different cut-off ELISA IgG antibody levels where the immunoblot was regarded as reference test. In total, 1454 patients were included. According to the two-step test strategy, 486 (33%), 69 (5%) and 899 (62%) patients had positive, indeterminate and negative Borrelia IgG serology, respectively. At IgG levels of 500 IU/ml and higher, all immunoblots were positive, resulting in a 100% PPV (95% CI: 97.0-100). At IgG levels of 200 IU/ml and higher, the PPV was 99.3% (95% CI: 97.4-99.8). In conclusion, at IgG levels of 200 IU/ml and higher, an ELISA was sufficient to detect antibodies to Borrelia burgdorferi s.l. At those IgG levels, a confirmatory immunoblot may be omitted in patients referred to a tertiary Lyme centre. Before these results can be implemented in routine diagnosis of Lyme borreliosis, confirmation of the results is necessary in other patient populations and using other quantitative ELISAs and immunoblots. Copyright © 2017 Elsevier GmbH. All rights reserved.

  12. Thermoglobin, oxygen-avid hemoglobin in a bacterial hyperthermophile.

    Science.gov (United States)

    Miranda, J J L; Maillett, David H; Soman, Jayashree; Olson, John S

    2005-11-04

    The hemoglobin family of proteins, ubiquitous in all domains of life, evolved from an ancestral protein of primordial function to extant hemoglobins that perform a myriad of functions with diverged biochemical properties. Study of homologs in bacterial hyperthermophiles may shed light on both mechanisms of adaptation to extreme conditions and the nature of the ancestral protein. A hemoglobin was identified in Aquifex aeolicus, cloned, recombinantly expressed, purified, and characterized. This hemoglobin is monomeric, resistant to thermal and chemical denaturation, pentacoordinate in the ferrous deoxygenated state, and oxygen-avid. The oxygen equilibrium dissociation constant is approximately 1 nm at room temperature, due in part to a hydrogen bond between the bound ligand and a tyrosine residue in the distal pocket. These biochemical properties of A. aeolicus thermoglobin, AaTgb, may have been shared by the ancestral hemoglobin, thus suggesting possible primordial functions and providing a starting point for consequent evolution of the hemoglobin family.

  13. Thallium-avid leiomyosarcoma of the chest wall: case report

    International Nuclear Information System (INIS)

    LaManna, M.M.; Lumia, F.J.; Feierstein, M.; Maranhao, V.

    1988-01-01

    A patient presented with leiomyosarcoma of the chest wall and parietal pleura, low grade malignant type, probably recurring from a section of a tumor of the parietal pleura diagnosed in 1979. During the course of pre-operative cardiac clearance, a resting thallium scan was performed. Examination of the study demonstrates dramatic thallium uptake in the left lung corresponding to the patient's lung mass. Extra cardiac accumulation of 201 Tl has been described in pulmonary neoplasms. The authors present an interesting case of a thallium-avid leiomyosarcoma of the chest wall which presented rather dramatic and discrete uptake of the isotope clearly distinguishable from the pulmonary activity which is seen left ventricular dysfunction, or low-level stress

  14. The use of ELISAs for monitoring exposure of pig herds to Brachyspira hyodysenteriae

    Directory of Open Access Journals (Sweden)

    Song Yong

    2012-01-01

    Full Text Available Abstract Background Swine dysentery (SD, a mucohaemorrhagic diarrhoeal disease of pigs, results from infection of the large intestine with the spirochaete Brachyspira hyodysenteriae. ELISA systems using whole spirochaete cells (WC and the B. hyodysenteriae outer membrane lipoprotein Bhlp29.7 previously have been established as potential diagnostic tools for SD. However, their true value in identifying infected herds remains unclear. The present study aimed to compare the performance of whole-cell and Bhlp29.7 based ELISAs in detecting specific immunoglobulin class IgG and IgM to B. hyodysenteriae in growing pigs, and additionally evaluated whether meat juice could serve as a source of specific antibodies. Results Levels of circulating IgG and IgM reacting with WC spirochaete preparations and recombinant Bhlp29.7 peaked 4-6 weeks post-infection in the experimentally challenged pigs, and remained elevated in the present study. In a cohort of pigs on an infected farm levels of antibody directed against both antigens showed a progressive increase with time. However, other than for the level of IgG against WC antigen, a significant increase in antibody levels also was observed in a cohort of pigs on a non-infected farm. In addition, assays using meat juice had 100% specificity and equivalent sensitivity to those based on serum, and likewise the best performance was achieved using the WC IgG ELISA. Conclusions IgG ELISAs using either WC or Bhlp29.7 as plate-coating antigens were shown to be useful for monitoring the dynamics of B. hyodysenteriae infection in grower pigs. Of the two antigens, the WC preparation tended to give better discrimination between pigs from infected and non-infected farms. Testing of meat juice was shown to have potential for identifying infected herds.

  15. Perbedaan Metode ELISA Sandwich A dan B dalam Deteksi Antigen Membran Toxoplasma gondii

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    ADI PRAYITNO

    2004-11-01

    Full Text Available Spreading of toxoplasmosis to fetus can by placenta, so it caused theabortion, born dead or congenital defect. To diagnosis this disease for fixed the acute infection must get the significant increasing of IgG by the soft fee. The objections of this study are to know the difference between ELISA Sandwich A and B in detecting of membrane antigen of Toxoplasma gondii (T. gondii in placenta tissue of pregnant women three-semester I and II with spontaneous abortion in Surakarta. One hundred serum and placenta tissue samples of pregnant women three-semester I and II with spontaneous abortion are got from dr. Muwardi Hospital. IgM anti Toxo from serum was examined by Toxo ISAGA Kit and IgG anti Toxo by Toxo Screen DA Kit. Detecting of membrane antigen of T. goodie from placenta tissue were done by ELISA Sandwich A and B. The result of this experiment showed that 33% were positive IBM and or Gig anti Toxo. Detection of membrane antigen toward 33 samples with positive Toxo (IgG positive was highly significant different between ELISA Sandwich A (3% positive toward ELISA Sandwich B (72.7% positive.

  16. Comparative evaluation of competitive ELISA test in Colombian cattle

    International Nuclear Information System (INIS)

    Marino, O.; Rueda, E.; Sedano, L.; Zuniga, I.; Calderon, C.; Ortega, A.; Puentes, A.

    1998-01-01

    In order to contribute to the definition of the best ELISA test for screening and differential diagnosis of Brucella abortus to be applied for control programmes, a total of 2971 sera from Colombian cattle were tested for brucellosis. Conventional agglutination tests, Buffered Plate antigen test (BPAT) and Rose Bengal (RB) as well as Complement Fixation test (CFT) (Alton, et al. 1988) were used comparatively. Radial immunodiffusion test (RID) was also performed to all sera. The sera were also tested using four different ELISAs: indirect ELISA from FAO/IAEA and the indirect ELISA modified by Nielsen, et al. 1992 as well as two competitive ELISAs: one competitive ELISA used B. abortus O-polysaccharide antigen and an enzyme conjugated monoclonal to the O-polysaccharide for competition and detection. The second competitive ELISA used lipopolysaccharide (sLPS) antigen, a different monoclonal antibody for competition but also specific for the O-polysaccharide and a commercially available goat anti-mouse IgG enzyme conjugate for detection. The sera were analyzed based on its population status, 987 positive obtained from Brucella abortus infected herds based on clinical and/or bacteriological evidence and a high prevalence of brucellosis, CFT percentage of positive animals in the herd was greater than 5%. Eight hundred sixty six (866) negative sera from non-vaccinated cattle from a brucellosis free area and 1118 negative sera obtained from reglamentary vaccinated areas under a free herd program. Initial cut-off values were derived using negative serum samples. The diagnostic sensitivity and specificity was defined from frequency histograms based on this cut-off values and using 2x2 tables, corresponding confidence limits (95%) were calculated. The data were also analysed using signal detection analysis (ROC). Kappa statistics was determined for all tests and populations, accuracy was used as index of comparison to evaluate different assays. The data support the initial

  17. The use of filter paper plasticized with polyvinyl alcohol-glutaraldehyde in ELISA

    Directory of Open Access Journals (Sweden)

    G.H.T.S. Barbosa

    2000-07-01

    Full Text Available F1-antigen purified from Yersinia pestis was covalently linked to 5-mm diameter filter paper discs plasticized with polyvinyl alcohol-glutaraldehyde. These discs were used both for ELISA and dot-ELISA for the detection of anti-F1 IgG in rabbits. The best conditions were achieved using 1.25 µg of F1 antigen/disc, 3% w/v skim milk in PBS as blocking agent, anti-IgG peroxidase conjugate diluted 12,000 times, and serum from rabbits immunized or not against Y. pestis, diluted 6,400 times. The absorbance values obtained from the comparative study between this procedure and conventional ELISA were not significantly different but the low cost of the reagents employed in ELISA using the filter paper discs plasticized with polyvinyl alcohol-glutaraldehyde makes this method economically attractive.

  18. ELISA for the detection of venom antigens in experimental and clinical envenoming by Loxosceles intermedia spiders.

    Science.gov (United States)

    Chávez-Olórtegui, C; Zanetti, V C; Ferreira, A P; Minozzo, J C; Mangili, O C; Gubert, I C

    1998-04-01

    Enzyme linked immunosorbent assays (ELISA) were developed to detect antigens from Loxosceles intermedia spider venom. Hyperimmune horse anti-Loxosceles intermedia IgGs were prepared by immunoaffinity chromatography and used to set up a sandwich-type ELISA. The specificity of the assay was demonstrated by its capacity to correctly discriminate the circulating antigens in mice that were experimentally inoculated with L. intermedia venom from those inoculated with L. gaucho, L. laeta, and Phoneutria nigriventer spider venoms, Tityus serrulatus scorpion venom and Bothrops jararaca, Crotalus durissus terrificus, Lachesis muta muta and Micrurus frontalis snake venoms. Measurable absorbance signals were obtained with 0.8 ng of venom per assay. The ELISA also detected antigens in the sera of patients envenomed by L. intermedia. Therefore, after standardization for clinical use this ELISA may be a valuable tool for clinicians and epidemiologists.

  19. Deficit of IgG2 in HIV-positive pregnant women is responsible of inadequate IgG2 levels in their HIV-uninfected children in Malawi.

    Science.gov (United States)

    Baroncelli, Silvia; Galluzzo, Clementina Maria; Liotta, Giuseppe; Andreotti, Mauro; Mancinelli, Sandro; Mphwere, Robert; Bokola, Enok; Amici, Roberta; Marazzi, Maria Cristina; Palombi, Leonardo; Lucaroni, Francesca; Giuliano, Marina

    2018-02-27

    Transplacental passage of IgGs is impaired in HIV + pregnant women, possibly determining an inadequate immunological protection in their children. We aimed to determine the impact of maternal immunological IgG profile and immunoactivation status on the efficiency of transplacental passage of IgG subclasses in HIV + mothers. 16 mother/infants pairs were studied in Malawi. Mothers received antiretroviral therapy (ART) from the third trimester of pregnancy. Determinations of pre-ART levels of maternal sCD14, of IgG subclasses in mothers at delivery and in their 1-month-old infants, were performed using commercial ELISA kits. At delivery, after a median of 10 weeks of ART, 12/16 mothers were hypergammaglobulinemic, with IgG levels (20.5 mg/ml, 95% CI:18.8-26.8) directly correlated to the plasmatic levels of sCD14 (r = 0.640, p = 0.014). IgG1 levels (17.9 mg/ml) accounted for 82% of IgG, IgG3 and IgG4 levels were in the normal range. A profound deficit of IgG2 was observed both in mothers (0.60 mg/ml) and in infants (0.14 mg/ml). Placental transfer ratio (range 0.16-0.42) did not show a selective impairment between the different IgG subclasses. The transplacental passage of all IgG subclasses was decreased in the presence of maternal IgG over 16 mg/ml (significantly for IgG1, p = 0.031) and of high levels of sCD14 (p = 0.063). Transplacental passage was reduced for all IgG subclasses and inversely correlated to high levels of maternal IgGs and to the degree of immunoactivation. The profound depression of IgG2 in mothers suggests that IgG2 neonatal levels mostly reflect the maternal deficit rather than a selective impairment of IgG2 transfer.

  20. The Diagnostic Value of ELISA Method for Pertussis in Children

    Directory of Open Access Journals (Sweden)

    O. P. Popova

    2016-01-01

    Full Text Available Because of low effectiveness of laboratory methods for diagnosing pertussis it is important to look for new ways of verification of this infection. The article presents the analysis of the diagnostic value of ELISA method, which involves the identification of antibodies of different isotypes (IgM, IgG, IgA to pertussis toxoid (PT and filamentous haemagglutinin (FHA. The study included 279 children: 114 were under 1 year of age, 165 — older than 1 year. The pertussis was confirmed in 74.3 ± 2.6% of patients by using ELISA method. A significant proportion of seronegative patients (46.1 ± 6.2 per cent was revealed in the group of patients under 1 year. The pattern of production of antibodies in unvaccinated children was different. It depended on the age of the children and timing of illness. A low proportion of diagnostically significant indicators of IgM-antibodies at 2—3 weeks of illness was typical for patients under 1 year of age (e.g. 6.7 ± 6.5% as compared to 20.0 ± 7.9% and 50.0 ± 15.3 — 1—3 and 4—6 years of age. The diagnosis of pertussis in children under 1 year of age was confirmed mainly by the detection of IgG, starting from the 4th week of the disease. In the examination of vaccinated children diagnostically significant levels of IgA and IgG were identified (even in the late stages of the disease. Thus, the results of the analysis show special significance of using ELISA method for the diagnosis of pertussis in vaccinated children.

  1. Serum IgG antibodies against Wallemia sebi and Fusarium species in Finnish farmers.

    Science.gov (United States)

    Lappalainen, S; Pasanen, A L; Reiman, M; Kalliokoski, P

    1998-12-01

    Wallemia sebi and Fusarium species are common fungi in agricultural environments. Because Fusarium species are difficult to culture, and W. sebi has only recently been found in an agricultural environment with the use of a new culture medium, immunochemical methods may be important for evaluating exposure to these fungi. Immunoglobulin (Ig) G antibodies against W. sebi and two Fusarium species were measured in the sera of Finnish farmers to estimate exposure to fungi. Serum-specific IgG antibody levels against the fungi were determined in farmers with farmer's lung disease and asthma, as well as in asymptomatic farmers with low and high IgG antibody levels against other agricultural microorganisms, and in control persons (printing and office workers). Both enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunofiltration assay (ELIFA) techniques were used. Significantly higher IgG antibody levels against the fungi studied were observed in farmer's lung patients and asymptomatic farmers with high IgG levels against other agricultural fungi, than in the control group. On the other hand, the development of IgG antibodies to the fungi in asthmatic farmers remained unclear because of a discrepancy between the ELISA and ELIFA results. Cross-reactivity between Fusarium species was detected. The results indicate that Finnish farmers are exposed to W. sebi and Fusarium species in their work environment to a greater extent than has earlier been evaluated with air sampling and microbiologic methods.

  2. Gold nanoparticle-based colorimetric ELISA for quantification of ractopamine.

    Science.gov (United States)

    Han, Shuaijuan; Zhou, Tianjiao; Yin, Bingjie; He, Pingli

    2018-03-07

    The work describes a gold nanoparticle-based colorimetric enzyme-linked immunosorbent assay (ELISA) for ractopamine. The ELISA is based on an indirect competitive approach. In the presence of ractopamine, gold(III) ions are oxidized by H 2 O 2 to form red AuNPs. On the other hand, the AuNP in solution are purple-blue due to aggregation if the sample does not contain ractopamine. The absorption, best measured at 560 nm, increases linearly in the 2 to 512 ng·mL -1 ractopamine concentration range, and the detection limit is as low as 0.35 ng·mL -1 in urine. Ractopamine can also be detected visually, even in the presence of other β-agonists and antibiotics. The results obtained by this method are consistent with those obtained by LC-MS/MS as demonstrated by analysis of sheep urine. The ELISA method described here is inexpensive, easy-to-use, and suitable for rapid screening of ractopamine in animal samples. Graphical abstract Schematic presentation of a colorimetric indirect competitive immunoassay for ractopamine. It is based on the use of catalase labeled IgG and the measurement of the absorption of red gold nanoparticles (AuNPs) that are generated by the reaction of gold ions with H 2 O 2 . In the absence of ractopamine, the solution becomes blue.

  3. Short communication: ELISA system for screening of bovine mastitis caused by Prototheca zopfii.

    Science.gov (United States)

    Kano, Rui; Sato, Ayano; Sobukawa, Hideto; Sato, Yuko; Ito, Takaaki; Suzuki, Kazuyuki; Hasegawa, Atsuhiko; Kamata, Hiroshi

    2016-08-01

    Prototheca zopfii is an achlorophyllic alga that causes bovine mastitis, resulting in a reduction in milk production and the secretion of thin, watery milk with white flakes. This study evaluated the use of an ELISA system for distinguishing cows with mastitis due to P. zopfii genotype 2 from healthy cows and cows with chronic candidal mastitis. We also investigated the transitional changes of specific antibody titers in healthy cows injected with inactivated P. zopfii genotype 2 cells. The ELISA system exhibited the highest sensitivity (94%) and specificity (100%) for chronic protothecal mastitis when the positive cutoff value was set at 43.4 ELISA units. Anti-protothecal IgG titers were positive in all cows after they were inoculated with inactivated P. zopfii genotype 2 cells. These results indicated that ELISA detection of anti-protothecal IgG in serum provided specificity and sensitivity sufficient for diagnosing protothecal mastitis. Thus, an ELISA system incorporating this specific antiserum is expected to be valuable for definitive field-based diagnosis of bovine mastitis due to P. zopfii genotype 2. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Development of purification processes for fully human bispecific antibodies based upon modification of protein A binding avidity.

    Science.gov (United States)

    Tustian, Andrew D; Endicott, Christine; Adams, Benjamin; Mattila, John; Bak, Hanne

    2016-01-01

    There is strong interest in the design of bispecific monoclonal antibodies (bsAbs) that can simultaneously bind 2 distinct targets or epitopes to achieve novel mechanisms of action and efficacy. Multiple bispecific formats have been proposed and are currently under development. Regeneron's bispecific technology is based upon a standard fully human IgG antibody in order to minimize immunogenicity and improve the pharmacokinetic profile. A single common light chain and 2 distinct heavy chains combine to form the bispecific molecule. One of the heavy chains contains a chimeric Fc sequence form (called Fc*) that ablates binding to Protein A via the constant region. As a result of co-expression of the 2 heavy chains and the common light chain, 3 products are created, 2 of which are homodimeric for the heavy chains and one that is the desired heterodimeric bispecific product. The Fc* sequence allows selective purification of the FcFc* bispecific product on commercially available affinity columns, due to intermediate binding affinity for Protein A compared to the high avidity FcFc heavy chain homodimer, or the weakly binding Fc*Fc* homodimer. This platform requires the use of Protein A chromatography in both a capture and polishing modality. Several challenges, including variable region Protein A binding, resin selection, selective elution optimization, and impacts upon subsequent non-affinity downstream unit operations, were addressed to create a robust and selective manufacturing process.

  5. AVIDITY EVALUATION OF LOCAL IgA ANTIBODIES IN PERSONS IMMUNIZED WITH LIVE INFLUENZA VACCINE

    Directory of Open Access Journals (Sweden)

    S. A. Donina

    2008-01-01

    Full Text Available Abstract. At present, immunogenicity evaluation of influenza vaccines is performed by quantitative assessment of increased serum antibodies. It was, however, shown that the degree of human defense against influenza is mostly related to their qualitative characteristics, i.e., avidity (functional activity. Leading role of local immunity is demonstrated in protection against influenza. Such immunity is mediated by IgA antibodies from mucosal airways. Meanwhile, the avidity issues for local antibodies still remain open.In present study, an attempt was undertaken to evaluate post-vaccination local immunological memory for influenza A virus, according to IgA antibodies from upper respiratory secretions. Two techniques were used to evaluate antibody avidity, that were previously applied for studying this phenomenon with serum imunoglobulins, i.e., a dynamic test (measurement of antigen-antibody reaction rates, and a test with urea, a chaotropic agent (avidity is determined as a strength of antigen-antibody complex. A total of 202 persons (18 to 20 years old were enrolled into the study.With both tests, a broad range of individual avidity values was observed for the antibodies. A significant cohort (up to 30 per cent of persons immunized with live influenza vaccine, showed sharply increased avidity of secretory IgA antibodies by both methods, along with accumulation of these immunoglobulins after vaccination. A reverse relationship is revealed between avidity levels of these antibodies before vaccination, and increase of this parameter post-immunization. The data present convincing arguments for specific renewal of local humoral immunological memory, as induced by live influenza vaccine. The study substantiates a necessity for application of the both tests in parallel, when determining avidity of secretory IgA antibodies. (Med. Immunol., vol. 10, N 4-5, pp 423-430.

  6. IgG4-related disease

    DEFF Research Database (Denmark)

    Detlefsen, Sönke; Klöppel, Günter

    2018-01-01

    disease (IgG4-RD). The histologic key findings are lymphoplasmacytic infiltration rich in IgG4-positive plasma cells combined with storiform fibrosis and obliterative phlebitis. Among the organs mainly affected by IgG4-RD are the pancreas and the extrahepatic bile ducts. The pancreatic and biliary...... alterations have been described under the terms autoimmune pancreatitis (AIP) and sclerosing cholangitis, respectively. These diseases are currently more precisely called IgG4-related pancreatitis (or type 1 AIP to distinguish it from type 2 AIP that is unrelated to IgG4-RD) and IgG4-related sclerosing...... cholangitis (IgG4-related SC). Clinically and grossly, both diseases commonly imitate pancreatic and biliary adenocarcinoma, tumors that are well known for their dismal prognosis. As IgG4-RD responds to steroid treatment, making a resection of a suspected tumor unnecessary, a biopsy is often required...

  7. Detection of pemphigus autoantibodies by IIF and ELISA tests in patients with pemphigus vulgaris and foliaceus and in healthy relatives

    NARCIS (Netherlands)

    Torzecka, Jolanta Dorota; Narbutt, Joanna; Sysa-Jedrzejowska, Anna; Waszczykowska, Elzbieta; Lukamowicz, Jolanta; Pas, Hendri H.

    2003-01-01

    BACKGROUND: Pemphigus is a life-threatening, autoimmune blistering disease, mediated by IgG autoantibodies. The aim of our study was to assess the usefulness of a new enzyme-linked immunosorbent assay (ELISA) in detecting circulating pemphigus autoantibodies, and to compare its sensitivity and

  8. Anti-rubella, Mumps and Measles IgG Antibodies in Medical Students of Tehran University.

    Science.gov (United States)

    Keshavarz, Maryam; Nicknam, Mohammad Hossein; Tebyanian, Majid; Shahkarami, Mohammad Kazem; Izad, Maryam

    2016-06-01

    Measles, mumps and rubella are viral infectious diseases that may result in serious complications. Since the production of vaccines, the number of cases of these diseases has been dropped. Nevertheless, these infectious diseases are still one of the major health problems in developing countries. In this study, in order to evaluate the protective responses against measles, mumps and rubella, the level and avidity of virus-specific IgG antibodies were measured in 53 medical students of Tehran University, aged between 20-30 years. Except for mumps vaccine, all the students had been vaccinated against measles and rubella according to Iran's nationwide mass vaccination protocol for all persons aged 5-25 in 2003. Our results showed that 96.2% of the volunteers had a protective level (>15 IU/ml) of IgG against rubella, 79.2% had a protective level (>11 IU/ml) of IgG against measles and 64.16% had a protective level (>11 IU/ml) of IgG against mumps. Over ten years after nationwide measles-rubella vaccination campaign, most young adults aged 20-30 had protective levels of humoral immunity against measles and rubella. However, Iranian young population is still unvaccinated against mumps, and therefore relatively large number of young adults had no protective level of IgG against it. This finding may be due to reduction in circulating of wild strain. We recommend screening of medical students for immunity against infectious agents such as measles, mumps, rubella, because they are at a high risk of these infectious agents.

  9. Sandwich ELISA Using a Mouse/Human Chimeric CSLEX-1 Antibody.

    Science.gov (United States)

    Yamashita, J; Kobayashi, I; Tatematsu, K; Sezutsu, H; Noda, K; Ishihara, H

    2016-11-01

    An assay using a mouse antisialyl Lewis X (sLeX) antibody (CSLEX-1) is used clinically for screening and monitoring patients with breast cancer in Japan. However, the IgM isoform of CSLEX-1 is not preferred for the assay because the bulkiness of IgM generally causes poor accessibility to the antigen. To solve this problem, we developed an antisLeX mouse/human chimeric IgG antibody, CH-CSLEX-1, using transgenic silkworms. The performance of a homologous sandwich ELISA of CH-CSLEX1 was then evaluated. To generate CH-CSLEX-1, we used a GAL4/UAS binary gene expression system in transgenic silkworms. The reactivities of CSLEX-1 and CH-CSLEX-1 were determined in a Biacore analysis. To confirm antigen specificity, 3 antigens [sLeX, sLeA, and Lewis Y (LeY)] were used. CH-CSLEX-1 formed correctly as an IgG class of immunoglobulin molecule with an isoelectric point close to the predicted value. The best combination for capturing and probing in a sandwich ELISA was determined as a homologous combination of CH-CSLEX-1. The CH-CSLEX-1 assay specifically detected sLeX, but not sLeA and LeY. A correlation analysis with 107 human samples showed good concordance between the conventional CSLEX-1 assay (homologous sandwich ELISA using CSLEX-1) and the CH-CSLEX-1 assay (r = 0.98). Moreover, the CH-CSLEX-1 assay was not affected by either human antimouse IgG antibodies (HAMA IgG) or HAMA IgM. The mouse/human chimeric antibody CH-CSLEX-1 allowed the establishment of a highly specific sandwich ELISA for sLeX that was not affected by HAMA. © 2016 American Association for Clinical Chemistry.

  10. Development and primary application of ELISA method for detecting residual calf serum protein

    International Nuclear Information System (INIS)

    Li Hua; Yu Jiankun; Hong Chao; Long Ruixiang; Zhai Yougang; Xu Qiongfang; Cui Pingfang; Ding Xuefeng; Xie Zhongping

    2005-01-01

    To develop a new DAS-ELISA (double antibody sandwich ELISA) kit for detecting residual calf serum protein (CSP) in vaccines, calf sera from different district were pooled and used to immunize rabbits and hens respectively. Then, the IgY from yolk and the anti-CSP IgG from rabbit were separated and purified. The purified IgY was used as the coating antibody, and purified rabbit anti-CSP IgG was labeled by HRP. The optimal concentration of IgY was 25-30 μg/mL. The coating buffer was 0.01mol/L PBS(pH7.4) containing 0.4% glutin. The optimal dilutions of HRP-IgG were from 1:2000 to 1:3000. The sensitivity of this ELISA method was higher (up to 2.5μg/mL) than that of current RPHA, the variation coefficient was about 6.3%-9.4%, and the recovery rate was 90.4%-112.8%. Furthermore, there was no cross-reaction with sera of pig, monkey and guinea pig. Twenty lots of vaccines with Al(OH) 3 or without Al(OH) 3 were tested by ELISA and RPHA respectively. The results proved that the adjuvant of Al(OH) 3 had fewer influence on ELISA than on RPHA, the variation of PRHA among different lots of vaccines was more significant than ELISA. The ELISA method is a highly sensitive and useful method to detect CSP in vaccines. (authors)

  11. Elisa technology consolidation study overview

    Science.gov (United States)

    Fitzsimons, E. D.; Brandt, N.; Johann, U.; Kemble, S.; Schulte, H.-R.; Weise, D.; Ziegler, T.

    2017-11-01

    The eLISA (evolved Laser Interferometer Space Antenna) mission is an ESA L3 concept mission intended to detect and characterise gravitational radiation emitted from astrophysical sources [1]. Current designs for eLISA [2] are based on the ESA study conducted in 2011 to reformulate the original ESA/NASA LISA concept [3] into an ESA-only L1 candidate named NGO (New Gravitational Observatory) [4]. During this brief reformulation period, a number of significant changes were made to the baseline LISA design in order to create a more costeffective mission. Some of the key changes implemented during this reformulation were: • A reduction in the inter satellite distance (the arm length) from 5 Gm to 1 Gm. • A reduction in the diameter of the telescope from 40 cm to 20 cm. • A reduction in the required laser power by approximately 40%. • Implementation of only 2 laser arms instead of 3. Many further simplifications were then enabled by these main design changes including the elimination of payload items in the two spacecraft (S/C) with no laser-link between them (the daughter S/C), a reduction in the size and complexity of the optical bench and the elimination of the Point Ahead Angle Mechanism (PAAM), which corrects for variations in the pointing direction to the far S/C caused by orbital dynamics [4] [5]. In the run-up to an L3 mission definition phase later in the decade, it is desirable to review these design choices and analyse the inter-dependencies and scaling between the key mission parameters with the goal of better understanding the parameter space and ensuring that in the final selection of the eLISA mission parameters the optimal balance between cost, complexity and science return can be achieved.

  12. Snake venomics of monocled cobra (Naja kaouthia) and investigation of human IgG response against venom toxins

    DEFF Research Database (Denmark)

    Laustsen, Andreas Hougaard; Gutiérrez, José María; Lohse, Brian

    2015-01-01

    /cardiotoxins. IgGs isolated from a person who had repeatedly self-immunized with a variety of snake venoms were immunoprofiled by ELISA against all venom fractions. Stronger responses against larger toxins, but lower against the most critical α-neurotoxins were obtained. As expected, no neutralization potential...

  13. Canine specific ELISA for coagulation factor VII

    DEFF Research Database (Denmark)

    Knudsen, Tom; Kjelgaard-Hansen, Mads; Tranholm, Mikael

    2011-01-01

    available to date. In this study, a canine specific ELISA for measurement of FVII:Ag in plasma was developed and validated. The FVII:Ag ELISA correctly diagnosed homozygous and heterozygous hereditary FVII deficiency. Together with activity based assays, such as FVII:C, the FVII:Ag ELISA should be valuable...

  14. Vaccination for birch pollen allergy. Induction of affinity-matured or blocking IgG antibodies does not account for the reduced binding of IgE to Bet v 1

    DEFF Research Database (Denmark)

    Svenson, Morten; Jacobi, Henrik H; Bødtger, Uffe

    2003-01-01

    the amount of Bet v 1 compared with IgE, and together they accounted for more than 85% of the serum binding capacity. While SAV induced minimal changes in IgE binding, the IgG binding capacities increased 6-32 times. In contrast, the binding avidities (K(d) 28-40pM) changed less than 20%, pre- and post...

  15. [Evaluation of immunoglobulin G concentration in colostrum of mares by ELISA, refractometry and colostrometry].

    Science.gov (United States)

    Venner, Monica; Markus, R G; Strutzberg-Minder, K; Nogai, K; Beyerbach, M; Klug, E

    2008-01-01

    In 360 samples of colostrum and 36 samples of blood of warmblood mares, the concentration of immunoglobulin G (IgG) was evaluated in the post partal period with an ELISA and the results were compared to values obtained with 2 field methods--refractometry and colostrometry. A significant correlation (p refractometry (r = +0.93). So both field-methods seem suitable for evaluation of the colostral IgG-concentration in mares. Further the kinetic of the IgG concentration in colostrum, the volume of colostrum and the total amount of IgG was measured in the 12 hours post partum (p.p.) in each half udder of 36 mares of different parity. Immediately p.p. primiparous mares have a greater mean concentration of IgG (68 mg/ml) than multiparous mares (51 mg/ml). However, multiparous mares have a mean colostral volume of 1020 ml whereas, in primiparous mares, a mean volume of 527 ml was determined within the first three hours p.p. As a result of this the total amount of IgG was lower in primiparous (31.5 g) than in multiparous mares (48.5 g). A significant decrease of IgG concentration was measured in multiparous mares in the 1.5 hours following partum versus 3 hours in primiparous mares. The mean IgG concentration in the blood serum of the 36 mares immediately p.p. was 13.4 +/- 3.6 mg/ml. No significant correlation was observed between values of IgG concentration in the blood and in the colostrum of the mares.

  16. Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA.

    Science.gov (United States)

    Tang, Qiushi; Gruntman, Alisha M; Flotte, Terence R

    2017-01-01

    In this chapter we describe an enzyme-linked immunosorbent assay (ELISA) to quantitatively measure human alpha-1 antitrypsin (AAT) protein levels in serum, other body fluids or liquid media. This assay can be used to measure the expression of the human AAT (hAAT) gene in a variety of gene transfer or gene downregulation experiments.A hAAT-specific capture antibody and a HRP-conjugated anti-AAT detection antibody are used in this assay. The conjugated anti-AAT used in this protocol, instead of the typical sandwich which employs an unconjugated antibody followed by a specifically conjugated IgG, makes the assay simpler and decreases variability. This provides a useful tool to evaluate the AAT levels in clinical and research samples and can allow fairly rapid testing of a large number of samples.

  17. Serum total IgG and IgG4 levels in thyroid eye disease

    Directory of Open Access Journals (Sweden)

    Sy A

    2016-10-01

    Full Text Available Aileen Sy, Rona Z Silkiss Department of Ophthalmology, California Pacific Medical Center, San Francisco, CA, USA Purpose: To investigate the relationship between immunoglobulin G (IgG4-related disease (IgG4-RD and thyroid eye disease (TED with respect to IgG levels. Patients and methods: A retrospective review of total IgG, IgG subclass, and thyroid stimulating immunoglobulin (TSI levels in 24 patients with TED. Results: Five patients (20.8% demonstrated serum IgG4 levels consistent with IgG4-RD without any additional systemic disease. Total IgG and IgG subclass levels were found to be an inadequate proxy for TSI elevation. Conclusion: There may be a subtype of TED patients with elevated IgG4 in the absence of IgG4-RD systemic findings. Keywords: thyroid eye disease, IgG subclass, IgG4, Graves’ disease, Graves’ ophthalmopathy, IgG4-RD

  18. A high boronate avidity monolithic capillary for the selective enrichment of trace glycoproteins.

    Science.gov (United States)

    Li, Daojin; Li, Yang; Li, Xinglin; Bie, Zijun; Pan, Xianghua; Zhang, Qian; Liu, Zhen

    2015-03-06

    Boronate affinity materials, as effective sample enrichment sorbents for glycoproteomic analysis, have attracted increasing attention in recent years. However, most of boronate affinity materials suffer from an apparent limitation, limited binding strength. As a result, extraction of glycoproteins of trace concentration is rather difficult or impossible. In this study, we present a high boronate avidity monolithic capillary. Branched polyethyleneimine (PEI) was used as a scaffold to amplify the number of boronic acid moieties. While 2,4-difluoro-3-formyl-phenylboronic acid (DFFPBA), which exhibited ultrahigh affinity toward cis-diol-containing compounds, was employed as an affinity ligand. Due to the PEI-assisted synergistic multivalent binding, the monolithic column exhibited high boronate avidity toward glycoproteins, with binding constants of 10(-6)-10(-7)M. Such binding strength was the highest among already reported boronic acid-functionalized materials that can be used for glycoproteomic analysis. Besides, the boronate avidity monolithic column exhibited one additional beneficial feature, lowered binding pH (≥6.5). These features greatly favored the selective enrichment of trace glycoproteins from real samples. The feasibility for practical applications was demonstrated with the selective enrichment of trace glycoproteins in human saliva. As compared with other boronate avidity/affinity materials, the boronate avidity monolithic capillary exhibited the best performance. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Serodiagnosis of toxocariasis by ELISA using crude antigen of Toxocara canis larvae.

    Science.gov (United States)

    Jin, Yan; Shen, Chenghua; Huh, Sun; Sohn, Woon-Mok; Choi, Min-Ho; Hong, Sung-Tae

    2013-08-01

    Toxocariasis is a worldwide zoonosis caused by larvae of ascarid nematodes of dogs or cats, Toxocara canis or T. cati. Diagnosis of human toxocariasis currently relies on serology that uses T. canis excretory-secretory antigen to detect specific IgG antibodies by ELISA. We investigated the serodiagnostic efficacy of ELISA using crude antigen of T. canis larvae (TCLA). Serum specimens of 64 clinically confirmed toxocariasis, 115 healthy controls, and 119 other tissue-invading helminthiases were screened by ELISA using TCLA. The ELISA using TCLA showed 92.2% (59/64 patient samples) sensitivity and 86.6% (103/119) specificity. Its positive diagnostic predictivity was 78.7% and negative predictivity was 97.8%. No serum of healthy controls reacted but that of anisakiasis (45.5%), gnathostomiasis (19.2%), clonorchiasis (15.8%), sparganosis (11.1%), and cysticercosis (6.3%) cross-reacted. Immunoblot analysis on TCLA recognized antigenic proteins of 28- and 30-kDa bands in their dominant protein quantity and strong blotting reactivity. The present results indicate that the ELISA using our TCLA antigen is acceptable by the sensitivity and specificity for serodiagnosis of human toxocariasis. ELISA with TCLA is recommended to make differential diagnosis for patients with any sign of organ infiltration and eosinophilia.

  20. [Application of food allergens specific IgG antibody detection in chronic diarrhea in children].

    Science.gov (United States)

    Ou-Yang, Wen-Xian; You, Jie-Yu; Duan, Bai-Ping; Chen, Chang-Bin

    2008-02-01

    The causes of chronic diarrhea in children are complex. At present, food allergy is generally viewed as an important cause of this disorder, and IgG-mediated delayed allergy plays a major role in this process. This study aimed to explore the link between food specific IgG and chronic diarrhea in children, as well as the value of food allergens-specific IgG antibody detection in the management of this disorder. Eighty-two children with chronic diarrhea and 30 healthy controls were enrolled. Serum levels of specific IgG antibody to 14 kinds of food were detected using ELISA. The results were classified into four grades: Grade 0 (negative), Grade 1 (mild allergy), Grade 2 (moderate allergy) and Grade 3 (severe allergy). The patients received a diet treatment based on the results of food specific IgG antibody detection. Children with negative IgG antibody were allowed to continue their current diet. In children with Grade 1 allergy, the food responsible for the IgG antibody positive test was given only at an interval of four days. In children with Grade 2 or 3, the offending food was eliminated from the diet. Of the 82 children with chronic diarrhea, 79 (96.2%) had increased specific IgG levels for one or more of the 14 foods tested compared to 8 (26.7%) of the controls (P chronic childhood diarrhea. Food specific IgG antibody detection may assist in the dietary management of this disorder.

  1. An enzyme-linked immunosorbent assay (ELISA) for methylphenidate (Ritalin ) in urine.

    Science.gov (United States)

    Lewis, Mark G; Lewis, John G; Elder, Peter A; Moore, Grant A

    2003-09-01

    A direct enyzme-linked immunosorbent assay (ELISA) for urinary immunoreactive methylphenidate (Ritalin), in which a standard 96-well microtiter plate is used, is described. For this ELISA, a methylphenidate-thyroglobulin conjugate is immobilized to the microtiter plate and competes with methylphenidate in the standard or urine sample for antibody-binding sites. After washing, the sheep methylphenidate antibody bound to immobilized methylphenidate is detected with peroxidase-labelled goat antisheep IgG. Following a further wash, tetramethylbenzidine is added, color is developed, and the plate is read at 450 nm on an ELISA plate reader. This method is unaffected by drugs of abuse and is suitable for routine use in the toxicology laboratory.

  2. Rapidly decreased serum IgG to Campylobacter pylori following elimination of Campylobacter in histological chronic biopsy Campylobacter-positive gastritis.

    Science.gov (United States)

    van Bohemen, C G; Langenberg, M L; Rauws, E A; Oudbier, J; Weterings, E; Zanen, H C

    1989-01-15

    The anaerobic bacterium Campylobacter pylori (Cp) is thought to be associated with chronic gastritis. This paper presents clinical data underpinning this view. Five patients with histological chronic gastritis as determined by diagnostic endoscopy, which was associated with Cp as determined by positive biopsy cultures, all possessed statistically raised serum IgG ELISA titers to Cp during a longitudinal period of observation of 15 months. Treatment with the antibiotics amoxycillin (clamoxyl) or colloidal bismuth subcitrate (denol) eliminated Cp within one month. Associated with this, serum IgG ELISA titers were found to decrease sharply and rapidly. Tagamet and spiramycin had little effect. Although the data are preliminary, they support the assumed Cp involvement in chronic gastritis and suggest that specific serum IgG ELISA titers to Cp are useful parameters in monitoring disease status, exceeding bacteriological culture of biopsy specimens in speed and convenience.

  3. Avidity of anti-circumsporozoite antibodies following vaccination with RTS,S/AS01E in young children.

    Directory of Open Access Journals (Sweden)

    Ally Olotu

    Full Text Available The nature of protective immune responses elicited by immunization with the candidate malaria vaccine RTS,S is still incompletely understood. Antibody levels correlate with protection against malaria infection, but considerable variation in outcome is unexplained (e.g., children may experience malaria despite high anti-circumsporozoite [CS] titers.We measured the avidity index (AI of the anti-CS antibodies raised in subgroup of 5-17 month old children in Kenya who were vaccinated with three doses of RTS,S/AS01E between March and August 2007. We evaluated the association between the AI and the subsequent risk of clinical malaria. We selected 19 cases (i.e., with clinical malaria and 42 controls (i.e., without clinical malaria, matching for anti-CS antibody levels and malaria exposure. We assessed their sera collected 1 month after the third dose of the vaccine, in March 2008 (range 4-10 months after the third vaccine, and at 12 months after the third vaccine dose. The mean AI was 45.2 (95% CI: 42.4 to 48.1, 45.3 (95% CI: 41.4 to 49.1 and 46.2 (95% CI; 43.2 to 49.3 at 1 month, in March 2008 (4-10 months, and at 12 months after the third vaccination, respectively (p = 0.9 by ANOVA test for variation over time. The AI was not associated with protection from clinical malaria (OR = 0.90; 95% CI: 0.49 to 1.66; p = 0.74. The AI was higher in children with high malaria exposure, as measured using the weighted local prevalence of malaria, compared to those with low malaria exposure at 1 month post dose 3 (p = 0.035.Our data suggest that in RTS,S/AS01E-vaccinated children residing in malaria endemic countries, the avidity of anti-circumsporozoite antibodies, as measured using an elution ELISA method, was not associated with protection from clinical malaria. Prior natural malaria exposure might have primed the response to RTS,S/AS01E vaccination.

  4. IgG subclass antibodies to human and bacterial HSP60 are not associated with disease activity and progression over time in axial spondyloarthritis

    DEFF Research Database (Denmark)

    Carlsen, Thomas Gelsing; Hjelholt, Astrid; Jurik, Anne Grethe

    2013-01-01

    in the same serum sample, IgG1 and IgG3, respectively, indicating that there was no cross-reaction. CONCLUSIONS: A significant association was observed between axial SpA and the presence of IgG1/IgG3 antibodies to human HSP60 and of IgG1 to S. enterica Enteritidis and C. trachomatis. Generation of antibodies...... and patient follow-up. In this study, we have focused on these parameters in a cohort of axial SpA patients with a well-established set of clinical characteristics, including MRI changes and human leukocyte antigen B27. METHODS: IgG subclass antibodies (IgG1, IgG2, IgG3 and IgG4) against recombinant HSP60...... of three reactive arthritis-related bacteria; human HSP60; and the microorganisms Chlamydia trachomatis and C. pneumoniae were determined by ELISA. Serum samples collected from 2004 to 2006 and in 2010 and 2011 from 39 axial SpA patients were analyzed and compared with samples from 39 healthy controls...

  5. Association between {sup 18}F-FDG avidity and the BRAF mutation in papillary thyroid carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Suk Hyun; Han, Sang Won; Lee, Hyo Sang; Chae, Sun Young; Lee, Jong Jin; Song, Dong Eun; Ryu, Jin Sook [Asan Medical Center, University of Ulsan College of Medicine, Seoul (Korea, Republic of)

    2016-03-15

    The BRAF mutation, a potential prognostic factor in papillary thyroid carcinoma (PTC), is associated with a high expression of the glucose transporter gene. We investigated which clinicopathologic factors, including BRAF mutation status, influence {sup 18}F-fluoro-2-deoxyglucose ({sup 18}F-FDG) avidity. We retrospectively reviewed 55 patients who underwent BRAF analysis from biopsy-confirmed PTC and {sup 18}F-FDG positron emission tomography/computed tomography within 6 months before undergoing thyroid surgery from September 2008 to August 2014. Tumors were considered to be {sup 18}F-FDG avid if the uptake was greater than that of the liver. {sup 18}F-FDG uptake of PTCs was also analyzed semiquantitatively using SUV{sub max}. The association between {sup 18}F-FDG avidity and clinicopathologic variables (age, tumor size, perithyroidal extension, cervical lymph node status, and BRAF mutation status) was investigated. Twenty-nine (52.7 %) of 55 patients had {sup 18}F-FDG-avid PTCs. PTCs with the BRAF mutation showed higher {sup 18}F-FDG avidity (24/38, 63.2 %) than those without (5/17, 29.4 %). The BRAF mutation (p = 0.025) and tumor size (p = 0.003) were significantly associated with {sup 18}F-FDG avidity in univariate analysis, and the BRAF mutation status remained significant after adjusting for tumor size in multivariate analysis (p = 0.015). In the subgroup of tumor size ≥ 1 cm, the BRAF mutation was the only factor significantly associated with {sup 18}F-FDG avidity (p = 0.021). The mean SUV{sub max} of PTCs with the BRAF mutation was significantly higher than that of those without (4.89 ± 6.12 vs. 1.96 ± 1.10, p = 0.039). The BRAF mutation must be one of the most important factors influencing {sup 18}F-FDG avidity in PTCs, especially in those with a tumor size ≥ 1 cm.

  6. Cutoff Values of Serum IgG4 and Histopathological IgG4+ Plasma Cells for Diagnosis of Patients with IgG4-Related Disease

    Directory of Open Access Journals (Sweden)

    Yasufumi Masaki

    2012-01-01

    Full Text Available IgG4-related disease is a new disease classification established in Japan in the 21st century. Patients with IgG4-related disease display hyper-IgG4-gammaglobulinemia, massive infiltration of IgG4+ plasma cells into tissue, and good response to glucocorticoids. Since IgG4 overexpression is also observed in other disorders, it is necessary to diagnose IgG4-related disease carefully and correctly. We therefore sought to determine cutoff values for serum IgG4 and IgG4/IgG and for IgG4+/IgG+ plasma cells in tissue diagnostic of IgG4-related disease. Patients and Methods. We retrospectively analyzed serum IgG4 concentrations and IgG4/IgG ratio and IgG4+/IgG+ plasma cell ratio in tissues of 132 patients with IgG4-related disease and 48 patients with other disorders. Result. Serum IgG4 >135  mg/dl demonstrated a sensitivity of 97.0% and a specificity of 79.6% in diagnosing IgG4-related disease, and serum IgG4/IgG ratios >8% had a sensitivity and specificity of 95.5% and 87.5%, respectively. IgG4+cell/IgG+ cell ratio in tissues >40% had a sensitivity and specificity of 94.4% and 85.7%, respectively. However, the number of IgG4+ cells was reduced in severely fibrotic parts of tissues. Conclusion. Although a recent unanimous consensus of all relevant researchers in Japan recently established the diagnostic criteria for IgG4-related disease, findings such as ours indicate that further discussion is needed.

  7. Serum IgG4 Concentration in IgG4-Related Disease.

    Science.gov (United States)

    Kawa, Shigeyuki; Skold, Markus; Ramsden, David B; Parker, Antony; Harding, Stephen J

    2017-09-01

    IgG4-related disease (IgG4-RD) is an immune-mediated and chronic fibroinflammatory condition that affects almost any organ and often involves multiple organs in the same patient. In this review article, we address the clinical utility of measuring serum immunoglobulin G subclass 4 concentration ([IgG4]) in IgG4-RD diagnosis and in disease monitoring. We discuss the latest literature on the relevance of [IgG4] to the investigation and management of IgG4RDs. In addition, we discuss the potential role of serum [IgG4] measurements in other inflammatory conditions and cancers. Increasing awareness of IgG4-RD among clinicians has led to a growing list of organ systems that can be affected by this chronic condition and the development of new organ-specific diagnostic guidelines. Diagnosis of IgG4-RD depends on multiple clinical and laboratory tests, including serology. Quantification of serum [IgG4] is included in all IgG4-RD diagnostic guidelines available to-date. The scientific literature supports the idea that elevated serum [IgG4], typically > 135 mg/dL, identifies patients with a more active form of the disease, which correlates with increased concentrations of inflammatory serum biomarkers and hypocomplementemia, increased number of organs affected by the disease, and more extensive organ involvement. These patients seem more resistant to treatment and experience a shorter time to disease relapse compared to IgG4-RD patients with normal serum [IgG4] at the time of diagnosis. Despite better understanding of how to diagnose IgG4-RD, monitoring for accurate prediction of disease relapse, which may involve organs not affected at the time of presentation, is poorly understood. Timely diagnosis and early detection of disease relapse is important to avoid delayed treatment and potential organ damage.

  8. Benign Bone Conditions That May Be FDG-avid and Mimic Malignancy

    NARCIS (Netherlands)

    Kwee, Thomas C.; de Klerk, John M. H.; Nix, Maarten; Heggelman, Ben G. F.; Dubois, Stefan V.; Adams, Hugo J. A.

    Positron emission tomography with the radiotracer F-18-fluoro-2-deoxy-D-glucose (FDG) plays an important role in the evaluation of bone pathology. However, FDG is not a cancer-specific agent, and knowledge of the differential diagnosis of benign FDG-avid bone alterations that may resemble malignancy

  9. Searching for avidity by chemical ligation of combinatorially self-assembled DNA-encoded ligand libraries.

    Science.gov (United States)

    Matysiak, Stefan; Hellmuth, Klaus; El-Sagheer, Afaf H; Shivalingam, Arun; Ariyurek, Yavuz; de Jong, Marco; Hollestelle, Martine J; Out, Ruud; Brown, Tom

    2017-12-19

    DNA encoded ligands are self-assembled into bivalent complexes and chemically ligated to link their identities. To demonstrate their potential as a combinatorial screening platform for avidity interactions, the optimal bivalent aptamer design (examplar ligands) for human alpha-thrombin is determined in a single round of selection and the DNA scaffold replaced with minimal impact on the final design.

  10. Porin A-specific antibody avidity in patients who are convalescing from meningococcal B disease.

    NARCIS (Netherlands)

    Vermont, C.L.; Dijken, H.H. van; Groot, R. de; Dobbelsteen, G.P. van den

    2005-01-01

    Porin A (PorA), which determines the serosubtype of Neisseria meningitidis, is the main antigen of a candidate vaccine against serogroup B meningococci, which has been shown to induce high-avidity antibodies in children. We characterized the immune response of children after convalescing from

  11. What Do Chaotrope-Based Avidity Assays for Antibodies to HIV-1 Envelope Glycoproteins Measure?

    Science.gov (United States)

    Alexander, Marina R; Ringe, Rajesh; Sanders, Rogier W; Voss, James E; Moore, John P; Klasse, Per Johan

    2015-06-01

    When HIV-1 vaccine candidates that include soluble envelope glycoproteins (Env) are tested in humans and other species, the resulting antibody responses to Env are sifted for correlates of protection or risk. One frequently used assay measures the reduction in antibody binding to Env antigens by an added chaotrope (such as thiocyanate). Based on that assay, an avidity index was devised for assessing the affinity maturation of antibodies of unknown concentration in polyclonal sera. Since a high avidity index was linked to protection in animal models of HIV-1 infection, it has become a criterion for evaluating antibody responses to vaccine candidates. But what does the assay measure and what does an avidity index mean? Here, we have used a panel of monoclonal antibodies to well-defined epitopes on Env (gp120, gp41, and SOSIP.664 trimers) to explore how the chaotrope acts. We conclude that the chaotrope sensitivity of antibody binding to Env depends on several properties of the epitopes (continuity versus tertiary- and quaternary-structural dependence) and that the avidity index has no simple relationship to antibody affinity for functional Env spikes on virions. We show that the binding of broadly neutralizing antibodies against quaternary-structural epitopes is particularly sensitive to chaotrope treatment, whereas antibody binding to epitopes in variable loops and to nonneutralization epitopes in gp41 is generally resistant. As a result of such biases, the avidity index may at best be a mere surrogate for undefined antibody or other immune responses that correlate weakly with protection. An effective HIV-1 vaccine is an important goal. Such a vaccine will probably need to induce antibodies that neutralize typically transmitted variants of HIV-1, preventing them from infecting target cells. Vaccine candidates have so far failed to induce such antibody responses, although some do protect weakly against infection in animals and, possibly, humans. In the search for

  12. Antigens of worms and eggs showed a differentiated detection of specific IgG according to the time of Schistosoma mansoni infection in mice

    Directory of Open Access Journals (Sweden)

    Rafaella Fortini Queiroz Grenfell

    2012-08-01

    Full Text Available INTRODUCTION: The correlation between the immunological assay and the antibody titer can offer a tool for the experimental analysis of different phases of the disease. METHODS: Two simple immunological assays for Schistosoma mansoni in mice sera samples based on specific IgG detection for worms soluble antigens and eggs soluble antigens were standardized and evaluated in our laboratory. Fifty mice were used in negative and positive groups and the results obtained by enzyme-linked immunosorbent assays (ELISA assays were compared with the number of worms counted and the IgG titers at different times of infection. RESULTS: Data showed that ELISA using adult worm antigens (ELISA-SWAP presented a satisfactory correlation between the absorbance value of IgG titers and the individual number of worms counted after perfusion technique (R²=0.62. In addition, ELISA-SWAP differentially detected positive samples with 30 and 60 days post infection (p=0.011 and 0.003, respectively, whereas ELISA using egg antigens (ELISA-SEA detected samples after 140 days (p=0.03. CONCLUSIONS: These data show that the use of different antigens in immunological methods can be used as potential tools for the analysis of the chronological evolution of S. mansoni infection in murine schistosomiasis. Correlations with human schistosomiasis are discussed.

  13. Serum IgG galactosylation in psoriatic arthritis patients undergoing a biological treatment. A preliminary report

    Directory of Open Access Journals (Sweden)

    Anna Jakubiak-Augustyn

    2014-11-01

    Full Text Available Aim of the study : A group of psoriatic arthritis patients (PSA, 11 cases has undergone a treatment. For each patient two serum samples were collected: before the treatment (sample before and after the treatment (sample after. Performed investigation aimed to provide an information whether a treatment of the PSA patients improves the galactosylation of IgG N-glycans, which previously was shown for rheumatoid arthritis (RA patients. Material and methods : Immunoglobulin G (IgG was isolated from serum of PSA patients, before and after the treatment, using affinity chromatography on Protein A-Sepharose, and was further analyzed regarding the galactose content, using gas chromatography-mass spectrometry (GC-MS method and ELISA test, performed with two lectins: Ricinus communis (RCA-I and Griffonia simplicifolia (GSL-II. Based on ELISA results an agalactosylation factor (AF was calculated for each before and after IgG sample. Results: Obtained data regarded galactose content in IgG, AF calculation and estimation of two laboratory indices of inflammation: erythrocyte sedimentation rate (ESR and C-reactive protein (CRP concentration in serum. Based on statistical methods it was shown that average difference (d of after and before measures of AF for PSA patients was Med HL = –0.097 (95% CI: –0.49–0.25, which means that applied treatment did not affect AF value for this group of patients in a statistically significant manner (p = 0.2936. Conclusions: The study demonstrated that, in the contrary to RA patients, IgG galactosylation in PSA patients, undergoing a treatment, showed no statistically significant changes.

  14. Optimization of a paper-based ELISA for a human performance biomarker.

    Science.gov (United States)

    Murdock, Richard C; Shen, Li; Griffin, Daniel K; Kelley-Loughnane, Nancy; Papautsky, Ian; Hagen, Joshua A

    2013-12-03

    Monitoring aspects of human performance during various activities has recently become a highly investigated research area. Many new commercial products are available now to monitor human physical activity or responses while performing activities ranging from playing sports, to driving, and even sleeping. However, monitoring cognitive performance biomarkers, such as neuropeptides, is still an emerging field due to the complicated sample collection and processing, as well as the need for a clinical lab to perform analysis. Enzyme-linked immunosorbent assays (ELISAs) provide specific detection of biomolecules with high sensitivity (picomolar concentrations). Even with the advantage of high sensitivity, most ELISAs need to be performed in a laboratory setting and require around 6 h to complete. Transitioning this assay to a platform where it reduces cost, shortens assay time, and is able to be performed outside a lab is invaluable. Recently developed paper diagnostics provide an inexpensive platform on which to perform ELISAs; however, the major limiting factor for moving out of the laboratory environment is the measurement and analysis instrumentation. Using something as simple as a digital camera or camera-enabled Windows- or Android-based tablets, we are able to image paper-based ELISAs (P-ELISAs), perform image analysis, and produce response curves with high correlation to target biomolecule concentration in the 10 pM range. Neuropeptide Y detection was performed. Additionally, silver enhancement of Au NPs conjugated with IgG antibodies showed a concentration-dependent response to IgG, thus eliminating the need for an enzyme-substrate system. Automated image analysis and quantification of antigen concentrations are able to be performed on Windows- and Android-based mobile platforms.

  15. Measurement of allergen-specific IgG in serum is of limited value for the management of dogs diagnosed with cutaneous adverse food reactions.

    Science.gov (United States)

    Hagen-Plantinga, E A; Leistra, M H G; Sinke, J D; Vroom, M W; Savelkoul, H F J; Hendriks, W H

    2017-02-01

    Conflicting results have been reported in the literature in terms of the usefulness of serological testing for IgG against food allergens in dogs with cutaneous adverse food reaction (CAFR). The aim of the present study was to evaluate the suitability of a commercially available IgG ELISA for identifying food allergens in dogs, by challenging dogs with specific food ingredients, selected on the basis of IgG reactivity in serum samples. A total of 24 adult dogs with CAFR were enrolled into the study and 16 healthy dogs were included as a control group. Blood samples were obtained for measurement of specific IgG antibodies against 39 commonly used pet food ingredients by ELISA. Participating owners were surveyed to obtain information on their pet's dietary history. Eleven healthy control dogs and 12 dogs with CAFR were subsequently challenged in a blinded cross-over design experiment with both positive and negative food ingredients, selected on the basis of the ELISA test results. There was substantial individual variation in ELISA test results to the various food allergens, but no significant difference in IgG reactivity comparing the CAFR and control groups. None of the control dogs developed any clinical signs of an allergic reaction during the dietary challenge study. In the CAFR group, six of 12 dogs developed clinical signs after the negative challenge, and two of nine dogs developed clinical signs after the positive challenge. It was concluded that the ELISA test for dietary allergen-specific IgG is of limited value in the management of dogs with CAFR. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. The fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi additionally binds IgG and contributes to virulence in a mouse model.

    Science.gov (United States)

    Meehan, M; Lynagh, Y; Woods, C; Owen, P

    2001-12-01

    The major cell-wall-associated protein of the equine pathogen Streptococcus equi subsp. equi is an M-like fibrinogen-binding protein (FgBP) which binds equine fibrinogen (Fg) avidly, through residues located at the extreme N-terminus of the molecule. In this study, it is shown that FgBP additionally binds equine IgG-Fc. When tested against polyclonal IgG from ten other animal species, it was found that FgBP binds human, rabbit, pig and cat IgG, but does not bind mouse, rat, goat, sheep, cow or chicken IgG. Through the use of a panel of recombinant FgBP truncates containing defined deletions of sequence, it was shown that residues in the central regions of FgBP are important in IgG binding. An fbp knockout mutant which does not express FgBP on the cell surface was also constructed. Mutant cells failed to autoaggregate, bound no detectable equine Fg or IgG-Fc, were rapidly killed in horse blood, and showed greatly decreased virulence in a mouse model. Results suggest that FgBP is the major surface structure responsible for binding either Fg or IgG, that the molecule has pronounced antiphagocytic properties, and that it is a likely factor contributing to the virulence of wild-type S. equi subsp. equi.

  17. Altered antibody response to influenza H1N1 vaccine in healthy elderly people as determined by HI, ELISA, and neutralization assay

    NARCIS (Netherlands)

    Remarque, E.J.; Bruijn, de I.A.; Boersma, W.J.A.; Masurel, N.; Ligthart, G.J.

    1998-01-01

    To determine the influence of ageing per se as well as of priming histories on the antibody response to influenza vaccination, haemagglutination inhibition (HI), ELISA IgG, IgA, IgM and neutralizing antibody titres were studied in 43 healthy young subjects (mean age 23 years) and 55 healthy elderly

  18. IgG4 and IgE co-positive group found in idiopathic orbital inflammatory disease.

    Science.gov (United States)

    Zhao, Peng-Xiang; Adzavon, Yao Mawulikplimi; Ma, Jian-Min; Shang, Lei; Chen, Dan-Ying; Xie, Fei; Liu, Meng-Yu; Zhang, Xin; Lyu, Bao-Bei; Zhang, Ming-Zi; Yang, Lin-Qi; Ma, Xue-Mei

    2018-01-01

    To reveal the cytokines involved in idiopathic orbital inflammatory disease (IOID) and the relationship between Th17 cells, IgE and IOID pathogenesis. Whole blood samples were processed immediately after collection and serological IgG4, IgG, and IgE antibodies were tested using ELISA. IOID and orbital cavernous hemangioma (CH) tissue samples underwent Bio-Plex multiplex cytokine detection. Hematoxylin-Eosin (HE) staining of all paraffin samples suggested the histological features of IOIDs, and expressions of IgG4 and IL-17A in affected tissues were detected by immunohistochemistry. Among 40 IOID plasma samples, 52.5% (21/40) were positive for IgG4 and 25% (10/40) were positive for IgE. Overlapped IgG4 or IgE positive samples accounted for 22.5% (9/40). Therefore, IOID samples were separated into three groups. The IgE+/IgG4+ group had a relevantly lower level of pro-inflammatory cytokine expression. IL-4 (Th2 cell related), IL-10 and TGF-β1 (Treg cell immunity related) were elevated in all three groups. Some of the Th17 cell related cytokines ( i.e. IL-17A/F, IL-25, IL-23, and IL-33) displayed higher expression levels in the IgE-/IgG4- group compared to the other two groups. We discovered an IgG4-IgE co-positive group as well as Th17 cell immune involvement in IgG4-IgE co-negative subgtroup in IOID for the first time. The pathogenesis of IOID could differ from different subgroups according to the IgG4 and IgE detection. Therefore, we recommend that, Treatment stratagy should be made according to the clinical assessment of IgG4-IgE and Th17 profile detection.

  19. Evaluation of a Newly Developed ELISA Against Widal, TUBEX-TF and Typhidot for Typhoid Fever Surveillance

    Science.gov (United States)

    2011-01-01

    Company, Sparks, Maryland, USA) using BD-Difco Salmonella flagellar d (TH), BD-Difco Salmonella O group D and Brucella abortus antigens...illness patients in Egypt. Am J Trop Med Hyg 77: 390-392. 14. Memish ZA, Almuneef M, Mah MW, Qassem LA, Osoba AO (2002) Comparison of the Brucella ...standard agglutination test with the ELISA IgG and IgM in patients with Brucella bacteremia. Diagn Microbiol Infect Dis 44: 129-132. 15. Fadeel MA

  20. Rapid and simple IgG specific test for the exclusion of heparin induced thrombocytopenia (HIT).

    Science.gov (United States)

    Kolde, Hans-Jürgen; Dostatni, Ralf; Mauracher, Susanne

    2011-08-29

    The exclusion of heparin induced thrombocytopenia (HIT) is required for selecting the most appropriate anticoagulation therapy in affected patients. It requires the combination of clinical data with the detection of antibodies directed against platelet factor 4 (PF4) in complex with polyanions (PA) such as heparin. We developed a lateral flow immunoassay (LFIA) for PF4/PA complex specific IgG antibodies based on gold nanoparticles. Unlike most other assays, the initial immune reaction takes place in the liquid phase. The sensitivity of the assay has been adjusted with clinical samples aiming in the reliable detection of sera which are positive in a functional platelet activation assay. Sera from 60 patients with suspected HIT were investigated. LFIA identified correctly all samples (n=20) which were positive in a functional assay (HIPA) and an IgG specific ELISA. It correlated with ELISA, but false positive results were less frequent (7 samples were negative with LFIA and HIPA but positive with ELISA). The LFIA may be a suitable tool for the rapid exclusion of HIT within 10 min.

  1. FDG avidity and PET/CT patterns in primary gastric lymphoma

    Energy Technology Data Exchange (ETDEWEB)

    Radan, Lea [Rambam Health Care Campus, Department of Nuclear Medicine, Haifa (Israel); Fischer, Doron [Rambam Health Care Campus, Department of Diagnostic Imaging, Haifa (Israel); Bar-Shalom, Rachel; Israel, Ora [Technion - Israel Institute of Technology, Department of Nuclear Medicine, Rambam Health Care Campus and R. and B. Rappaport School of Medicine, Haifa (Israel); Dann, Eldad J. [Technion - Israel Institute of Technology, Department of Hematology, Rambam Health Care Campus, and R. and B. Rappaport School of Medicine, Haifa (Israel); Epelbaum, Ron; Haim, Nissim [Technion - Israel Institute of Technology, Department of Oncology, Rambam Health Care Campus, and R. and B. Rappaport School of Medicine, Haifa (Israel); Gaitini, Diana [Rambam Health Care Campus, and R. and B. Rappaport School of Medicine, Technion - Israel Institute of Technology, Department of Diagnostic Imaging, Haifa (Israel)

    2008-08-15

    The use of 18F-fluoro-deoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) in primary gastric lymphoma (PGL) is challenging due to physiologic FDG activity in the stomach and variability in the degree of uptake in various histologic subtypes. This study assesses FDG avidity and PET/CT patterns in newly diagnosed PGL. Sixty-two PET/CT studies of newly diagnosed PGL were reviewed (24 low-grade mucosa-associated lymphoid tissue [MALT], 38 aggressive non-Hodgkin's lymphoma [AGNHL]). FDG avidity, patterns (focal/diffuse), and intensity (visually vs. the liver and SUVmax) were assessed and compared to 27 controls. Gastric CT abnormalities and extragastric sites were recorded. Gastric FDG uptake was found in 55/62 (89%) PGL (71% MALT vs. 100% AGNHL, p < 0.001) and 63% controls. A diffuse pattern was found in 60% PGL (76% MALT vs. 53% AGNHL, p = NS) and 47% controls. FDG uptake higher than liver was found in 82% PGL (58% MALT vs. 97% AGNHL, p < 0.05) and 63% controls. SUVmax in FDG-avid PGLs was 15.3 {+-} 11.7 (5.4 {+-} 2.9 MALT vs. 19.7 {+-} 11.5 AGNHL, p < 0.001) and 4.6 {+-} 1.4 in controls. CT abnormalities were found in 79% PGL (thickening, n = 49; ulcerations, n = 22). Extra-gastric FDG-avid sites were seen in none of MALT, but 61% of AGNHL (nodal, n = 18; nodal and extranodal, n = 5). FDG avidity was present in 89% of PGLs, including all patients with AGNHL but only 71% of MALT. FDG uptake can be differentiated, in particular in AGNHL-PGL, from physiologic tracer activity by intensity but not by pattern. Extragastric foci on PET and structural CT abnormalities are additional parameters that can improve PET/CT assessment of PGL. Defining FDG avidity and PET/CT patterns in AGNHL and a subgroup of MALT-PGL before treatment may be important for further monitoring therapy response. (orig.)

  2. High avidity antibodies to full-length VAR2CSA correlate with absence of placental malaria.

    Directory of Open Access Journals (Sweden)

    Yeung Lo Tutterrow

    Full Text Available VAR2CSA mediates sequestration of Plasmodium falciparum-infected erythrocytes in the placenta, increasing the risk of poor pregnancy outcomes. Naturally acquired antibodies (Ab to placental parasites at delivery have been associated with improved pregnancy outcomes, but Ab levels and how early in pregnancy Ab must be present in order to eliminate placental parasites before delivery remains unknown. Antibodies to individual Duffy-binding like domains of VAR2CSA have been studied, but the domains lack many of the conformational epitopes present in full-length VAR2CSA (FV2. Thus, the purpose of this study was to describe the acquisition of Ab to FV2 in women residing in high and low transmission areas and determine how Ab levels during pregnancy correlate with clearance of placental parasites. Plasma samples collected monthly throughout pregnancy from pregnant women living in high and low transmission areas in Cameroon were evaluated for Ab to FV2 and the proportion of high avidity Ab (i.e., Ab that remain bound in the presence of 3M NH(4SCN was assessed. Ab levels and proportion of high avidity Ab were compared between women with placental malaria (PM(+ and those without (PM(- at delivery. Results showed that PM(- women had significantly higher Ab levels (p = 0.0047 and proportion of high avidity Ab (p = 0.0009 than PM(+ women throughout pregnancy. Specifically, women with moderate to high Ab levels (>5,000 MFI and those with ≥ 35% high avidity Ab at 5-6 months were found to have 2.3 (95% CI, 1.0-4.9 and 7.6-fold (p = 0.0013, 95% CI: 1.2-50.0 reduced risk of placental malaria, respectively. These data show that high levels of Ab to FV2, particularly those with high avidity for FV2, produced by mid-pregnancy are important in clearing parasites from the placenta. Both high Ab levels and proportion of high avidity Ab to FV2 may serve as correlates of protection for assessing immunity against placental malaria.

  3. Optimized expression of full-length IgG1 antibody in a common E. coli strain.

    Directory of Open Access Journals (Sweden)

    Conrad En Zuo Chan

    Full Text Available Multi-polypeptide proteins such as antibodies are difficult to express in prokaryotic systems such as E. coli due to the complexity of protein folding plus secretion. Thus far, proprietary strains or fermenter cultures have been required for appreciable yields. Previous studies have shown that expression of heterologous proteins in E. coli can be enhanced by the reduction of protein translation rates. In this paper, we demonstrate that useful quantities of full-length IgG can be expressed and purified from the common E. coli laboratory strain HB2151 in standard shaking culture using a simple strategy of reduced inducer concentration combined with delayed induction times to modulate translation rates. Purified IgG had only marginally reduced avidity compared to mammalian derived IgG. This indicates that this technique can be used to derive antibodies of potentially equal utility as those expressed in mammalian cell culture, particularly for applications where effector functions mediated by the glycosylated residues in the Fragment Crystallizable (Fc portion of the immunoglobulin are not required.

  4. The anti-IRBP IgG1 and IgG2a response does not correlate with susceptibility to experimental autoimmune uveitis

    Directory of Open Access Journals (Sweden)

    L. Vieira de Moraes

    2006-06-01

    Full Text Available Susceptibility to experimental autoimmune uveitis (EAU in inbred mice has been associated with a dominant Th1 response. Elevated anti-inter-photoreceptor retinoid-binding protein (anti-IRBP IgG2a/IgG1 antibody ratios have been implicated as candidate markers to predict disease severity. In the present study, both the anti-IRBP antibody isotype and severity of EAU phenotypes were examined in 4 non-isogenic genetically selected mouse lines to determine if they can be used as general markers of disease. Mice between 8 and 12 weeks old selected for high (H III or low (L III antibody response and for maximum (AIR MAX or minimum (AIR MIN acute inflammatory reaction (AIR were immunized with IRBP. Each experiment was performed with at least 5 mice per group. EAU was evaluated by histopathology 21 days after immunization and the minimal criterion was inflammatory cell infiltration of the ciliary body, choroid and retina. Serum IgG1- and IgG2a-specific antibodies were determined by ELISA. EAU was graded by histological examination of the enucleated eyes. The incidence of EAU was lower in AIR MIN mice whereas in the other strains approximately 40% of the animals developed the disease. Low responder animals did not produce anti-IRBP IgG2a antibodies or interferon-gamma. No correlation was observed between susceptibility to EAU and anti-IRBP isotype profiles. Susceptibility to EAU is related to the intrinsic capacity to mount higher inflammatory reactions and increased production of anti-IRBP IgG2a isotype is not necessarily a marker of this immunologic profile.

  5. Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

    NARCIS (Netherlands)

    Tran, Thanh Nga T.; de Vries, Peter J.; Hoang, Lan Phuong; Phan, Giao T.; Le, Hung Q.; Tran, Binh Q.; Vo, Chi Mai T.; Nguyen, Nam V.; Kager, Piet A.; Nagelkerke, Nico; Groen, Jan

    2006-01-01

    BACKGROUND: The reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients. METHODS: 781 pairs of acute (t0) and convalescent sera, obtained after three weeks (t3) and 161 corresponding pairs of filter paper blood spots were tested

  6. Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

    NARCIS (Netherlands)

    Tran, T.N.T.; de Vries, P.J.; Hoang, L.P.; Phan, G.T.; Le, H.Q.; Tran, B.Q.; Vo, C.M.T.; Nguyen, N.V.; Kager, P.A.; Nagelkerke, N.; Groen, J.

    2006-01-01

    Background: The reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients. Methods: 781 pairs of acute (t0) and convalescent sera, obtained after three weeks (t3) and 161 corresponding pairs of filter paper blood spots were tested

  7. Desarrollo de un ensayo casero para la detección de IgG contra el core del virus de la Hepatitis B

    Directory of Open Access Journals (Sweden)

    Yismelvy Márquez-Toro

    2016-12-01

    Full Text Available Hepatitis B infection affects individuals worldwide, especially in Latin America. Serological assay for HBV antibodies and antigen detection, are critical for HBV diagnostic and treatment. Available commercial ELISA kits are expensive and in our country not always are readily available. The aim of this study was develop a homemade ELISA kit for serological detection IgG anti-HBV core (IgG anti-HBc accessible for our patients. 114 samples were analyzed: 17 from seronegatives individuals, 48 from seropositives patients and 49 belonging to indigenous population from Mérida State. Exposure to HBV was determined using the Murex anti-HBc test (DiaSorin, UK. Homemade ELISA shows 100% specificity and del 100 % sensitivity, and was in very good agreement with serological status, Kappa= 1 (CI 95%: 0,767-1, X2 (p<0,001. In indigenous population (Wayuu and mestizos, overall IgG anti-HBc prevalence was 22%. Taking into account these results, homemade IgG anti-HBc ELISA is efficacious and low cost usefully, for HBV diagnosis.

  8. Characteristics of enzyme-linked immunosorbent assay for detection of IgG antibodies specific to Сhlamydia trachomatis heat shock protein (HSP-60

    Directory of Open Access Journals (Sweden)

    O. Yu. Galkin

    2017-02-01

    Full Text Available The goal of this work was to study sensitivity and specificity of the developed ELISA set for the identification of IgG antibodies against Chlamydia trachomatis HSP-60 (using biotinylated tyramine-based signal amplification system. The study was conducted using a panel of characterized sera, as well as two reference ELISA sets of similar purpose. According to the results of ELISA informative value parameters, the ELISA we have developed showed the highest specificity and sensitivity parameters (no false negative or false positive results were registered. In 4 out of 15 intralaboratory panel serum samples initially identified as negative, anti-HSP-60 IgG-antibodies test result in reference ELISA sets upon dilution changed from negative to positive. The nature of titration curves of false negative sera and commercial monoclonal antibodies А57-В9 against C. trachomatis HSP-60 after incubation for 24 h was indicative of the presence of anti-idiotypic antibodies in these samples. Upon sera dilution, idiotypic-anti-idiotypic complexes dissociated, which caused the change of test result. High informative value of the developed ELISA set for identification of IgG antibodies against C. trachomatis HSP-60 has been proven. Anti-idiotypic antibodies possessing C. trachomatis anti-HSP-60 activity and being one of the causes of false negative results of the relevant ELISA-based tests have been identified in blood sera of individuals infected with chlamydial genitourinary infection agents.

  9. An ELISA for detection of apoptosis.

    OpenAIRE

    Salgame, P; Varadhachary, A S; Primiano, L L; Fincke, J E; Muller, S; Monestier, M

    1997-01-01

    We describe a simple and convenient enzyme-linked immunosorbent assay (ELISA) for the detection of apoptosis in tissue culture. An early event in apoptosis is DNA fragmentation followed by release of nucleosomes into the cytoplasm. Our sandwich assay uses a pair of monoclonal antibodies specific for two nucleosomal epitopes to capture and detect cytoplasmic nucleosomes onto the ELISA plate. Our assay is about 500 times more sensitive than the detection of apoptotic DNA ladder by agarose elect...

  10. Field trial of brucellosis competitive ELISA

    International Nuclear Information System (INIS)

    Perez, B.; Rojas, M.

    1998-01-01

    2990 sera samples from cattle were tested for antibodies to Brucella abortus using 8 serological tests for. The tests used were Rose Bengal (RBT), Buffer Plate Agglutination Test (BPAT), Complement Fixation (CFT), 2 Indirect and 2 Competitive Enzyme Linked Immunosorbent Assays (ELISA). Bacteriological evaluation from milk was done also. All tests were compared with respect to diagnostic specificity in vaccinated herds which were considered to be Brucella-free. The diagnostic specificity of the Indirect and Competitive ELISA was greater than 99,8%. Estimates of relative sensitivity were obtained from infected herds. The diagnostic sensitivity of the Indirect ELISA was greater than 95,8% and for the Competitive ELISA between 98,8 and 100 %, the last value refers to the Competitive ELISA Prototype II (SLPS antigen/M84 Mab), which was found highly suitable to differentiate vaccinated from brucella-infected cattle. The use of C-ELISA II for monitoring bovine populations under an eradication programme is recommended. (author)

  11. [The IgG antibody response in patients colonized by Helicobacter pylori].

    Science.gov (United States)

    Figueroa, G; Acuña, R; Jashés, M; Troncoso, M; Toledo, M S; Arellano, L

    1990-11-01

    The IgG antibody response specific to Helicobacter pylori was evaluated through ELISA in a group of 92 gastric patients colonized by this bacteria. 74 had gastritis and 19 gastroduodenal ulcer. Three control groups were studied in a similar way: normal adult volunteers (n = 17), adults with E coli or S typhi bacteremia (n = 30) and normal infants (n = 30). IgG antibody response to H pylori was demonstrated in 98% of colonized patients and 0% of infants. Asymptomatic individuals and those with bacteremia had high rates of antibody response (76 and 90% respectively), although this rate and also the titers of antibody response were significantly lower than that of colonized patients (p pylori in the majority of colonized gastric patients and asymptomatic adults suggests that this infection is very common in our population.

  12. Radiometallated receptor-avid peptide conjugates for specific in vivo targeting of cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Hoffman, T.J.; Quinn, T.P.; Volkert, W.A. E-mail: VolkertW@health.missouri.edu

    2001-07-01

    New receptor-avid radiotracers are being developed for site-specific in vivo targeting of a myriad of receptors expressed on cancer cells. This review exemplifies strategies being used to design radiometallated peptide conjugates that maximize uptake in tumors and optimize their in vivo pharmacokinetic properties. Efforts to produce synthetic peptide analogues that target the following three receptor systems are highlighted: Gastrin releasing peptide (GRP), alpha-melanocyte stimulating hormone ({alpha}-MSH), and guanylate cyclase-C (GC-C) receptors.

  13. Avidity Modulation of Folate-Targeted Multivalent Dendrimers for Evaluating Biophysical Models of Cancer Targeting Nanoparticles

    Science.gov (United States)

    Silpe, Justin E.; Sumit, Madhuresh; Thomas, Thommey P.; Huang, Baohua; Kotlyar, Alina; van Dongen, Mallory A.; Banaszak Holl, Mark M.; Orr, Bradford G.; Choi, Seok Ki

    2013-01-01

    We investigated two types of generation 5 polyamidoamine (PAMAM) dendrimers, each conjugated stochastically with a mean number of five or ten methotrexate (MTX) ligands per dendrimer (G5-MTX5, G5-MTX10), for their binding to surface-immobilized folate binding protein (FBP) as a function of receptor density. The binding study was performed under flow by surface plasmon resonance spectroscopy. Two multivalent models were examined to simulate binding of the dendrimer to the receptor surface, showing that at relatively high receptor density, both dendrimer conjugates exhibit high avidity. However, upon reducing the receptor density by a factor of three and thirteen relative to the high density level, the avidity of the lower-valent G5-MTX5 decreases by up to several orders of magnitude (KD = nM to μM), whereas the avidity of G5-MTX10 remains largely unaffected regardless of the density variation. Notably, on the 13-fold reduced FBP surface, G5-MTX5 displays binding kinetics similar to that of monovalent methotrexate, which is patently different from the still tight binding of the higher-valent G5-MTX10. Thus, the binding analysis demonstrates that avidity displayed by multivalent MTX conjugates varies in response to the receptor density, and it can be modulated for achieving tighter, more specific binding to the higher receptor density by modulation of ligand valency. We believe this study provides experimental evidence supportive of the mechanistic hypothesis of multivalent NP uptake to a cancer cell over a healthy cell where the diseased cell expresses the folate receptor at higher density. PMID:23855478

  14. Intratypic differentiation of poliovirus strains by enzyme-linked immunosorbent assay (ELISA): poliovirus type 1.

    Science.gov (United States)

    Glikmann, G; Moynihan, M; Petersen, I; Vestergaard, B F

    1983-01-01

    A double antibody sandwich-ELISA has been developed for the detection of antigenic differences between wild and vaccine derived strains of Poliovirus type 1. Poliovirus strains antibodies were prepared in rabbits by immunization with virus suspensions of: Sabin LSc2ab (vaccine derived) and Brunhilde and Mahoney (wild types). IgG fractions were purified from antiserum by precipitation with ammonium sulphate and DEAE-Sephadex A50 chromatography. Purified IgG antibodies were used for coating of microtest plates (catching antibodies). The same reagents labeled with horseradish peroxidase were used as conjugates (detecting antibodies). Detecting antibodies were made strain specific by cross-absorption with the heterologous virus strain. Absorbed and non-absorbed detecting antibodies were subsequently used for detection and quantitation of the poliovirus antigen(s) bound to IgG-coated surfaces. Poliovirus laboratory strains and isolates from sixty-six individuals were differentiated intratypically as vaccine derived or wild types when the ELISA was performed using absorbed conjugates. No intermediate strains were found, and all clinical samples tested fell in two distinct categories. Conversely, when detecting antibodies were used before absorption a high degree of homology between wild and vaccine strains was demonstrated and the differentiation between the two groups was poorly achieved. The ELISA has been optimized in terms of specificity and sensitivity. Less than 10 ng of poliovirus antigens could be detected by non-absorbed detecting antibodies whereas 18 ng was the minimal amount detected by the same antibodies after absorption. Preparation of strain specific antibodies did not require a previous concentration of the poliovirus suspension used for the absorption. It is proposed that the developed ELISA is capable of: 1) detection of low amounts of poliovirus antigens in clinical samples, and 2) intratypic differentiation of poliovirus antigens as either vaccine

  15. Investigating the role of T-cell avidity and killing efficacy in relation to type 1 diabetes prediction.

    Directory of Open Access Journals (Sweden)

    Anmar Khadra

    2011-05-01

    Full Text Available During the progression of the clinical onset of Type 1 Diabetes (T1D, high-risk individuals exhibit multiple islet autoantibodies and high-avidity T cells which progressively destroy beta cells causing overt T1D. In particular, novel autoantibodies, such as those against IA-2 epitopes (aa1-577, had a predictive rate of 100% in a 10-year follow up (rapid progressors, unlike conventional autoantibodies that required 15 years of follow up for a 74% predictive rate (slow progressors. The discrepancy between these two groups is thought to be associated with T-cell avidity, including CD8 and/or CD4 T cells. For this purpose, we build a series of mathematical models incorporating first one clone then multiple clones of islet-specific and pathogenic CD8 and/or CD4 T cells, together with B lymphocytes, to investigate the interaction of T-cell avidity with autoantibodies in predicting disease onset. These models are instrumental in examining several experimental observations associated with T-cell avidity, including the phenomenon of avidity maturation (increased average T-cell avidity over time, based on intra- and cross-clonal competition between T cells in high-risk human subjects. The model shows that the level and persistence of autoantibodies depends not only on the avidity of T cells, but also on the killing efficacy of these cells. Quantification and modeling of autoreactive T-cell avidities can thus determine the level of risk associated with each type of autoantibodies and the timing of T1D disease onset in individuals that have been tested positive for these autoantibodies. Such studies may lead to early diagnosis of the disease in high-risk individuals and thus potentially serve as a means of staging patients for clinical trials of preventive or interventional therapies far before disease onset.

  16. MIBG avidity correlates with clinical features, tumor biology, and outcomes in neuroblastoma: A report from the Children's Oncology Group.

    Science.gov (United States)

    DuBois, Steven G; Mody, Rajen; Naranjo, Arlene; Van Ryn, Collin; Russ, Douglas; Oldridge, Derek; Kreissman, Susan; Baker, David L; Parisi, Marguerite; Shulkin, Barry L; Bai, Harrison; Diskin, Sharon J; Batra, Vandana; Maris, John M; Park, Julie R; Matthay, Katherine K; Yanik, Gregory

    2017-11-01

    Prior studies suggest that neuroblastomas that do not accumulate metaiodobenzylguanidine (MIBG) on diagnostic imaging (MIBG non-avid) may have more favorable features compared with MIBG avid tumors. We compared clinical features, biologic features, and clinical outcomes between patients with MIBG nonavid and MIBG avid neuroblastoma. Patients had metastatic high- or intermediate-risk neuroblastoma and were treated on Children's Oncology Group protocols A3973 or A3961. Comparisons of clinical and biologic features according to MIBG avidity were made with chi-squared or Fisher exact tests. Event-free (EFS) and overall (OS) survival compared using log-rank tests and modeled using Cox models. Thirty of 343 patients (8.7%) had MIBG nonavid disease. Patients with nonavid tumors were less likely to have adrenal primary tumors (34.5 vs. 57.2%; P = 0.019), bone metastases (36.7 vs. 61.7%; P = 0.008), or positive urine catecholamines (66.7 vs. 91.0%; P < 0.001) compared with patients with MIBG avid tumors. Nonavid tumors were more likely to be MYCN amplified (53.8 vs. 32.6%; P = 0.030) and had lower norepinephrine transporter expression. Patients with MIBG nonavid disease had a 5-year EFS of 50.0% compared with 38.7% for patients with MIBG avid disease (P = 0.028). On multivariate testing in high-risk patients, MIBG avidity was the sole adverse prognostic factor for EFS identified (hazard ratio 1.77; 95% confidence interval 1.04-2.99; P = 0.034). Patients with MIBG nonavid neuroblastoma have lower rates of adrenal primary tumors, bone metastasis, and catecholamine secretion. Despite being more likely to have MYCN-amplified tumors, these patients have superior outcomes compared with patients with MIBG avid disease. © 2017 Wiley Periodicals, Inc.

  17. TRIM5α SPRY/coiled-coil interactions optimize avid retroviral capsid recognition.

    Directory of Open Access Journals (Sweden)

    Marcin D Roganowicz

    2017-10-01

    Full Text Available Restriction factors are important components of intrinsic cellular defense mechanisms against viral pathogens. TRIM5α is a restriction factor that intercepts the incoming capsid cores of retroviruses such as HIV and provides an effective species-specific barrier to retroviral infection. The TRIM5α SPRY domain directly binds the capsid with only very weak, millimolar-level affinity, and productive capsid recognition therefore requires both TRIM5α dimerization and assembly of the dimers into a multivalent hexagonal lattice to promote avid binding. Here, we explore the important unresolved question of whether the SPRY domains are flexibly linked to the TRIM lattice or more precisely positioned to maximize avidity. Biochemical and biophysical experiments indicate that the linker segment connecting the SPRY domain to the coiled-coil domain adopts an α-helical fold, and that this helical portion mediates interactions between the two domains. Targeted mutations were generated to disrupt the putative packing interface without affecting dimerization or higher-order assembly, and we identified mutant proteins that were nevertheless deficient in capsid binding in vitro and restriction activity in cells. Our studies therefore support a model wherein substantial avidity gains during assembly-mediated capsid recognition by TRIM5α come in part from tailored spacing of tethered recognition domains.

  18. Synthetic Cancer-Targeting Innate Immune Stimulators Give Insights into Avidity Effects.

    Science.gov (United States)

    Conibear, Anne C; Pötgens, André J G; Thewes, Karine; Altdorf, Claudia; Hilzendeger, Clarissa; Becker, Christian F W

    2018-03-02

    Multispecific and multivalent antibodies are seen as promising cancer therapeutics, and numerous antibody fragments and derivatives have been developed to exploit avidity effects that result in increased selectivity. Most of these multispecific and multivalent antibody strategies make use of recombinant expression of antigen-binding modules. In contrast, chemical synthesis and chemoselective ligations can be used to generate a variety of molecules with different numbers and combinations of binding moieties in a modular and homogeneous fashion. In this study we synthesized a series of targeted immune system engagers (ISErs) by using solid-phase peptide synthesis and chemoselective ligations. To explore avidity effects, we constructed molecules bearing different numbers and combinations of two "binder" peptides that target ephrin A2 and integrin α 3 receptors and an "effector" peptide that binds to formyl peptide receptors and stimulates an immune response. We investigated various strategies for generating multivalent and multispecific targeted innate immune stimulators and studied their activities in terms of binding to cancer cells and stimulation of immune cells. This study gives insights into the influence that multivalency and receptor density have on avidity effects and is useful for the design of potential anticancer therapeutics. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Dispensing an enzyme-conjugated solution into an ELISA plate by adapting ink-jet printers.

    Science.gov (United States)

    Lonini, Luca; Accoto, Dino; Petroni, Silvia; Guglielmelli, Eugenio

    2008-04-24

    The rapid and precise delivery of small volumes of bio-fluids (from picoliters to nanoliters) is a key feature of modern bioanalytical assays. Commercial ink-jet printers are low-cost systems which enable the dispensing of tiny droplets at a rate which may exceed 10(4) Hz per nozzle. Currently, the main ejection technologies are piezoelectric and bubble-jet. We adapted two commercial printers, respectively a piezoelectric and a bubble-jet one, for the deposition of immunoglobulins into an ELISA plate. The objective was to perform a comparative evaluation of the two classes of ink-jet technologies in terms of required hardware modifications and possible damage on the dispensed molecules. The hardware of the two printers was modified to dispense an enzyme conjugate solution, containing polyclonal rabbit anti-human IgG labelled with HRP in 7 wells of an ELISA plate. Moreover, the ELISA assay was used to assess the functional activity of the biomolecules after ejection. ELISA is a common and well-assessed technique to detect the presence of particular antigens or antibodies in a sample. We employed an ELISA diagnostic kit for the qualitative screening of anti-ENA antibodies to verify the ability of the dispensed immunoglobulins to bind the primary antibodies in the wells. Experimental tests showed that the dispensing of immunoglobulins using the piezoelectric printer does not cause any detectable difference on the outcome of the ELISA test if compared to manual dispensing using micropipettes. On the contrary, the thermal printhead was not able to reliably dispense the bio-fluid, which may mean that a surfactant is required to modify the wetting properties of the liquid.

  20. Chicken IgY-based coproantigen capture ELISA for diagnosis of human opisthorchiasis.

    Science.gov (United States)

    Teimoori, Salma; Arimatsu, Yuji; Laha, Thewarach; Kaewkes, Sasithorn; Sereerak, Piya; Sripa, Manop; Tangkawattana, Sirikachorn; Brindley, Paul J; Sripa, Banchob

    2017-08-01

    Diagnosis of Opisthorchis viverrini infection by conventional stool examination is increasingly difficult due to the low intensity of the infection after several rounds of control programmes in endemic regions as well as coinfections with intestinal flukes. Therefore sensitive and specific diagnostic test is needed. In this study, a coproantigen sandwich ELISA using recombinant O. viverrini cathepsin F (rOv-CF) was developed. This sandwich ELISA employing chicken IgY raised against rOv-CF in combination with rabbit IgG antibody to the somatic O. viverrini antigens showed a lower detection limit (LLD) of 70ng native O. viverrini somatic antigens by spiking the parasite antigens into control feces. When applied to the diagnosis, the IgY-based sandwich ELISA exhibited sensitivity and specificity of 93.3% and 76.7%, respectively, in an investigation of 90 human cases positive or negative for opisthorchiasis. The positive predictive value (PPV) and negative predictive value (NPV) for this coproantigen detection were 66.7% and 95.2%, respectively. This IgY-based sandwich ELISA using parasite cathepsin F detection shows a promising immunodiagnostic alternative for human opisthorchiasis in endemic regions. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. Combined use of Western blot/ELISA to improve the serological diagnosis of human tuberculosis

    Directory of Open Access Journals (Sweden)

    S. T. Beck

    Full Text Available Two recombinant antigens and a crude bacterial antigen of a wild M. tuberculosis strain were used to detect specific IgG antibodies in sera from 52 patients with pulmonary tuberculosis, confirmed by an acid-fast smear and serum culture of these patients and that of 25 contacts. The patients were not infected with HIV. We evaluated the sensitivity and specificity of ELISA, based on the recombinant TbF6® and TbF6/DPEP antigen and a search for reactivity patterns in the Western blot technique, using whole mycobacterium antigen. Serum samples from 22 healthy individuals and from 30 patients with lung diseases other than tuberculosis were used as controls. The best ELISA results were obtained with the TbF6/DPEP antigen combination, which gave 85% sensitivity and 91% specificity. ELISA sensitivity improved from 85% to 92% when the Western blot results were used. Western blot specificity was 100% when antibody reactivity with different antigenic bands was analyzed and associated. The association of TbF6/DPEP antigens used in ELISA with specific patterns of reactivity determined by Western blot can help make an identification when classic methods for the diagnosis of pulmonary tuberculosis are not sufficient.

  2. Combined use of Western blot/ELISA to improve the serological diagnosis of human tuberculosis.

    Science.gov (United States)

    Beck, Sandra Trevisan; Leite, O M; Arruda, R S; Ferreira, A W

    2005-02-01

    Two recombinant antigens and a crude bacterial antigen of a wild M. tuberculosis strain were used to detect specific IgG antibodies in sera from 52 patients with pulmonary tuberculosis, confirmed by an acid-fast smear and serum culture of these patients and that of 25 contacts. The patients were not infected with HIV. We evaluated the sensitivity and specificity of ELISA, based on the recombinant TbF6 and TbF6/DPEP antigen and a search for reactivity patterns in the Western blot technique, using whole mycobacterium antigen. Serum samples from 22 healthy individuals and from 30 patients with lung diseases other than tuberculosis were used as controls. The best ELISA results were obtained with the TbF6/DPEP antigen combination, which gave 85% sensitivity and 91% specificity. ELISA sensitivity improved from 85% to 92% when the Western blot results were used. Western blot specificity was 100% when antibody reactivity with different antigenic bands was analyzed and associated. The association of TbF6/DPEP antigens used in ELISA with specific patterns of reactivity determined by Western blot can help make an identification when classic methods for the diagnosis of pulmonary tuberculosis are not sufficient.

  3. Detection of IgG anti-beta 2 glycoprotein-I antibodies in Saudi patients with systemic lupus erythematosis

    International Nuclear Information System (INIS)

    Alenzi, Faris Q.

    2008-01-01

    Objective was to develop an assay for the measurement of this anti-human beta2-glycoprotein I. This study was conducted from September 2004 to December 2006. The patients attending the Rheumatology Clinic were chosen from several centers in the Eastern region of Saudi Arabia because they had complications. An enzyme-linked immunosorbent (ELISA) assay was optimized and developed to measure antibody levels in humans. Fifty normal blood donors and 50 systemic lupus erythrematosis (SLE) patients were selected for this experiment. Raised IGg a beta 2 GPI antibody levels were associated with the presence of venous thrombosis and thrombocytopenia. The real value of IgG a beta 2 GPI as a predictor for the future clinical complications needs to be confirmed in prospective controlled studies investigating clinical complications in relationship to IgG a beta 2 GPI and to other risk factors for thrombosis. (author)

  4. Natural anti-insulin autoantibodies in cats: enzyme-linked immunosorbent assay for the determination of plasma anti-insulin IgG and its concentrations in domestic cats.

    Science.gov (United States)

    Takashima, Satoshi; Nishii, Naohito; Hachisu, Tatsuyuki; Kojima, Masaaki; Kigure-Hoshino, Megumi; Ogawa, Shizuko; Suzuki, Takafumi; Iwasawa, Atsushi; Ohba, Yasunori; Kitagawa, Hitoshi

    2013-12-01

    Anti-insulin immunoglobulin G (IgG) has been found in the sera of healthy cats. To determine the concentrations of these antibodies, an enzyme-linked immunosorbent assay (ELISA) for anti-insulin IgG was developed. ELISA maintained the linearity of a standard concentration line between 67.5 and 2160 ng/ml. The coefficients of variances (CVs) of intra-assays in two different plasma samples were 4.0% and 3.7%, respectively. The inter-assay CVs in two different plasma samples were 5.1% and 6.9%, respectively. The dilution curves of two samples were rectilinear. Anti-insulin IgG was detected in all 84 of the healthy cats that were tested. Plasma anti-insulin IgG concentrations ranged from 80 to 1578 μg/ml, with a median concentration of 221 μg/ml, and this value correlated positively with total plasma IgG concentrations (r=0.383, pglucose tolerance test, plasma anti-insulin IgG concentrations did not alter, even with changes in plasma glucose and insulin concentrations. The ELISA that was developed was able to determine plasma anti-insulin IgG in domestic cats, and confirmed that all healthy cats had plasma anti-insulin IgG. Determining the plasma concentrations of anti-insulin IgG in cats with various pathological conditions might clarify the role of anti-insulin IgG. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. O blood group as a risk factor for Helicobacter pylori IgG seropositivity among pregnant Sudanese women

    Directory of Open Access Journals (Sweden)

    Gasim I. Gasim

    2017-06-01

    Full Text Available The objective was to investigate the prevalence and the association between blood groups and Helicobacter pylori IgG seropositivity among pregnant Sudanese women. A cross-sectional survey was carried- out at Saad Abul Ela Maternity Hospital, Khartoum, Sudan during the period of July 2014 through December 2015. Questionnaires covering socio-demographic and obstetrics information were administered. Specific H. pylori IgG antibody was analysed using ELISA. One hundred eighty six pregnant women were enrolled. The mean (SD of the age, parity was 28.3 (2.6 years and 2.6 (3.5, respectively. Of the 186 women, 42 (22.6%, 24 (12.9%, 11(5.9% and 109 (58.6% had blood group A, B, AB and O, respectively. H. pylori IgG seropositivity rate was 132/186 (71.0%. There was no significant difference in age and parity between women with H. pylori IgG seropositive and seronegative. Compared with the women with H. pylori IgG seronegative, significantly higher numbers of women with H. pylori IgG seropositive had O blood group, [84/132(63.6 versus 25/54(46.3, P<0.001]. In binary logistic regression, women with O blood group (OR= 2.084, 95% CI=1.060 -4.097, P=0.033 were at a higher H. pylori IgG seropositivity. The current study showed that women with blood group O were at higher risk for H. pylori IgG seropositivity.

  6. IgG4 related sclerosing mastitis: expanding the morphological spectrum of IgG4 related diseases.

    Science.gov (United States)

    Chougule, Abhijit; Bal, Amanjit; Das, Ashim; Singh, Gurpreet

    2015-01-01

    IgG4 related disease (IgG4RD) is a recently recognised condition characterised by mass forming lesions associated with storiform fibrosis, obliterative phlebitis, lymphoplasmacytic infiltrate rich in IgG4 positive plasma cells and elevated serum IgG4 levels. Although rare, mammary involvement has been reported as IgG4 related sclerosing mastitis, the morphological counterpart of a growing family of IgG4 related diseases. A total of 17 cases belonging to mass forming benign inflammatory breast lesions such as plasma cell mastitis, granulomatous lobular mastitis, non-specific mastitis and inflammatory pseudotumour were investigated as a possible member of IgG4 related sclerosing mastitis. Clinical, radiological, histopathological and immunohistochemistry findings were noted in all cases. Cases diagnosed as inflammatory pseudotumour showed all the histopathological features of IgG4RD along with increased number of IgG4 positive plasma cells and IgG4/IgG ratio >40%. However, only a few IgG4 positive cells were seen in plasma cell mastitis, granulomatous lobular mastitis and non-specific mastitis cases. These cases also did not fulfill the morphological criteria for the diagnosis of IgG4 related diseases. IgG4RD should be excluded in plasma cell rich lesions diagnosed on core biopsies by IgG4 immunostaining. This can avoid unnecessary surgery as IgG4 related diseases respond to simple and effective steroid treatment.

  7. Cholangiocarcinoma with respect to IgG4 Reaction

    Directory of Open Access Journals (Sweden)

    Kenichi Harada

    2014-01-01

    Full Text Available IgG4 reactions marked by infiltration of IgG4-positive plasma cells in affected organs occur in cancer patients and in patients with IgG4-related diseases. Extrahepatic cholangiocarcinomas including gall bladder cancer are often accompanied by significant IgG4 reactions; these reactions show a negative correlation with CD8-positive cytotoxic T cells, suggesting that the evasion of immune surveillance is associated with cytotoxic T cells. The regulatory cytokine IL-10 may induce IgG4-positive plasma cell differentiation or promote B cell switching to IgG4 in the presence of IL-4. Cholangiocarcinoma cells may function as nonprofessional antigen presenting cells that indirectly induce IgG4 reactions via the IL-10-producing cells and/or these may act as Foxp3-positive and IL-10-producing cells that directly induce IgG4 reactions. Moreover, IgG4-related disease is a high-risk factor for cancer development; IgG4-related sclerosing cholangitis (IgG4-SC cases associated with cholangiocarcinoma or its precursor lesion biliary intraepithelial neoplasia (BilIN have been reported. IgG4-positive cell infiltration is an important finding of IgG4-SC but is not a histological hallmark of IgG4-SC. For the diagnosis of IgG4-SC, its differentiation from cholangiocarcinoma remains important.

  8. Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis

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    Luciana Pereira Silva

    2003-07-01

    Full Text Available The present study was conducted to evaluate the frequency of antigenic components recognized by serum IgG antibodies in Western blotting (WB using a Strongyloides ratti larval extract for the diagnosis of human strongyloidiasis. In addition, the WB results were compared to the enzyme-linked immunosorbent assay (ELISA and the indirect immunofluorescence antibody test (IFAT results. Serum samples of 180 individuals were analyzed (80 with strongyloidiasis, 60 with other intestinal parasitoses, and 40 healthy individuals. S. ratti was obtained from fecal culture of experimentally infected Rattus rattus. For IFAT, S. ratti larvae were used as antigen and S. ratti larval antigenic extracts were employed in WB and ELISA. Eleven S. ratti antigenic components were predominantly recognized by IgG antibodies in sera of patients with strongyloidiasis. There was a positive concordance for the three tests in 87.5% of the cases of strongyloidiasis. The negative concordance in the three tests was 94% and 97.5%, in patients with other intestinal parasitoses and healthy individuals, respectively. In cases of positive ELISA and negative IFAT results, diagnosis could be confirmed by WB. ELISA, IFAT, and WB using S. ratti antigens showed a high rate of sensitivity and specificity. In conclusion, WB using S. ratti larval extract was able to recognize 11 immunodominant antigenic components, showing to be a useful tool to define the diagnosis in cases of equivocal serology.

  9. [IgG4-related disease].

    Science.gov (United States)

    González-Moreno, Juan; Losada López, Inés; Ortego Centeno, Norberto

    2015-12-21

    IgG4-related disease is a recently described clinicopathological entity showing a wide spectrum of clinical manifestations that share a common pathology. Its most characteristic feature is the formation of inflammatory tumors in different organs, which makes differentiation mainly with neoplastic diseases fundamental. The inflammatory process is typically comprised of IgG4 lymphoplasmacytic cells. The pathophysiological role of the immunoglobulin is not clear. The treatment of choice is corticosteroids. This article aims to summarize the main features of the disease. Copyright © 2015 Elsevier España, S.L.U. All rights reserved.

  10. Determination of Antibodies (IgG, IgM against Toxoplasma gondii in Patients with Cancer

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    M Pedram

    2007-08-01

    Full Text Available Background: The aim of this study was determination of antibodies (IgG, IgM against Toxoplasma in malignant patients in order to refer the patients on time to the physician for treatment.Methods: This study was carried out on 252 malignant patients and 252 healthy normal subjects (as control obtained from Shafa Hospital and Medical Diagnostic Laboratory (Iran-Zamin, in Ahwaz city. Patient's information was recorded in a questionnaire before sampling. Serum samples of patients were examined for IgG and IgM antibodies by ELISA technique using Trinity kits. Results: The results of this study revealed the presence of Toxoplasma antibodies in 114 (45.2% cases of patients who were positive for Toxoplasma IgG antibodies, and 26 (10.3% cases were confirmed to be positive for Toxoplasma IgM antibodies and also 17 (6.7% of cases had both IgG and IgM antibodies against Toxoplasma gondii. In control group 92 (36.5% cases and 15 (6% cases revealed seropositive for IgG and IgM antibodies, respectively. There were no significant differences between sex, close contact with cat, living region, chemotherapy, and seropositivity rate of toxoplasmosis in patients. Comparing the age groups, the highest seropositive rate showed in the age of 51 years or higher, and their rates had tendency to increase with age in both groups. No seropositivity significant relationship was found between patients and control group.Conclusion: According to the prevalence of positive cases in these patients, it is necessary to examine the patients for toxoplasmosis before, during and after chemotherapy.

  11. Seroprevalence of diphtheria toxoid IgG antibodies in children, adolescents and adults in Poland.

    Science.gov (United States)

    Zasada, Aleksandra A; Rastawicki, Waldemar; Rokosz, Natalia; Jagielski, Marek

    2013-11-19

    Recommendations for diphtheria immunization are to apply an effective primary immunization in infancy and to maintain immunity throughout life. Immunity against diphtheria depends primarily on antibody to the diphtheria toxin. This study evaluated the seroprevalence of IgG diphtheria antitoxin in sera of healthy children, adolescents and adults in Poland. A total of 1387 serum samples collected between 2010 and 2012 from individuals with ages ranging from 1 month to 85 years were investigated. Antibody concentrations were measured with an enzyme-linked immunosorbent assay (Anti-Diphtheria Toxoid ELISA IgG, Euroimmun, Germany). The results showed that among 1387 individuals examined, 547 (39.4%) had anti-diphtheria toxoid IgG antibody levels below 0.1 IU/ml (36.9% ≤ 18 years and 40.5% >18 years old, respectively). The 212 (50.8%) children and 542 (55.9%) adults showed only basic protection (0.1-1.0 IU/ml) and need immediate booster. High levels of anti-diphtheria toxoid IgG antibodies (>1.0 IU/ml) were found more often in children and adolescent (12.2%) than in adults (3.6%) and this was statistically significant (P 60 years old. Characteristically, in individuals > 40 years old high levels of anti-diphtheria toxoid IgG antibodies (>1.0 IU/ml) were not seen. There were no statistically significant differences in results in relation to gender. The present study showed inadequate immunity levels to diphtheria amongst the Polish population, especially in adults > 40 years old and children ≤ 2 years old. To prevent reemergence of diphtheria an information campaign reminding people about recommendations concerning diphtheria booster vaccination in adults should be conducted. Moreover, the immunogenicity of the DTP vaccine used in Poland should be verified.

  12. Estudo comparativo dos testes imunoenzimáticos elisa-g e elisa-m, imunofluorescência indireta e fixação do complemento no diagnóstico da cisticercose humana

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    Lucy G. Vianna

    1992-09-01

    Full Text Available Foi feito estudo comparativo entre quatro testes imunológicos - imunoenzimático IgG (ELISA-G e Ig M (ELISA-M, imunofluorescência indireta (RIFI e fixação do complemento (RFC - utilizados na detecção de anticorpos anti-Cysticercus, em soro e líquido ce-falomaquidiano (LCR de pacientes com suspeita clínica de cisticercose e seus familiares. Foram examinados 539 pacientes que apresentavam sintomas e/ou sinais sugestivos de cisticercose, 450 familiares destes doentes e 133 pessoas que constituíram o grupo controle. Foram colhidas 1122 amostras de soro e 120 de LCR que foram analisadas por ELISA-G e RIFI; em 83 soros e 60 LCR também foi processada a RFC e em 28 LCR também a ELISA-M. A ELISA-G e a RIFI mostraram-se reagentes em 5,2% dos soros, havendo discordância entre seus resultados em 3,5%. Em todos os soros do grupo controle ambos os testes foram não-reagentes. Estas mesmas reações, no LCR, foram reagentes em 16,7% e mostraram resultados discordantes em 7,5%. Houve concordância dos resultados da ELISA-G e aa RIFI, efetuadas concomitantemente no soro e no LCR, em 89,6% dos doentes, senão 17,7% reagentes. Nos soros em que foram executadas ELISA-G, RIFI e RFC, 54,2% mostraram concordância de resultados nos três testes, sendo reagentes em 16,9%. Estas mesmas reações no LCR tiveram resultados concordantes em 81,7%, sendo 11,7% reagentes. Nas amostras que apresentaram ELISA-G e RIFI nao-reagentes, a RFC foi reagente no soro e LCR, respectivamente, em 41,0% e 11,7%. Nos LCR em que se realizaram ELISA-G e ELISA-M, houve concordancia de resultados em 78,6%; nas amostras com resultados discordantes, 10,7% tiveram ELISA-G reagente e ELISA-M não-reagente, ocorrendo o inverso nas outras 10,7%. É dada ênfase à necessidade da realização concomitantemente de vários testes imunológicos para detecção de anticorpos anti-Cysticercus, no soro e no LCR, garantindo maior segurança no diagnóstico e acompanhamento evolutivo da doença.

  13. Milk protein IgG and IgA: The association with milk-induced gastrointestinal symptoms in adults

    Science.gov (United States)

    Anthoni, Sari; Savilahti, Erkki; Rautelin, Hilpi; Kolho, Kaija-Leena

    2009-01-01

    AIM: To study the association between serum levels of milk protein IgG and IgA antibodies and milk-related gastrointestinal symptoms in adults. METHODS: Milk protein IgG and IgA antibodies were determined in serum samples of 400 subjects from five outpatient clinics in Southern Finland. Subjects were randomly selected from a total of 1900 adults undergoing laboratory investigations in primary care. All 400 participants had completed a questionnaire on abdominal symptoms and dairy consumption while waiting for the laboratory visit. The questionnaire covered the nature and frequency of gastrointestinal problems, the provoking food items, family history and allergies. Twelve serum samples were disqualified due to insufficient amount of sera. The levels of specific milk protein IgG and IgA were measured by using the ELISA technique. The association of the milk protein-specific antibody level was studied in relation to the milk-related gastrointestinal symptoms and dairy consumption. RESULTS: Subjects drinking milk (n = 265) had higher levels of milk protein IgG in their sera than non-milk drinkers (n = 123, P < 0.001). Subjects with gastrointestinal problems related to milk drinking (n = 119) consumed less milk but had higher milk protein IgG levels than those with no milk-related gastrointestinal symptoms (n = 198, P = 0.02). Among the symptomatic subjects, those reporting dyspeptic symptoms had lower milk protein IgG levels than non-dyspeptics (P < 0.05). However, dyspepsia was not associated with milk drinking (P = 0.5). The association of high milk protein IgG levels with constipation was close to the level of statistical significance. Diarrhea had no association with milk protein IgG level (P = 0.5). With regard to minor symptoms, flatulence and bloating (P = 0.8), were not associated with milk protein IgG level. Milk protein IgA levels did not show any association with milk drinking or abdominal symptoms. The levels of milk protein IgA and IgG declined as the age

  14. Milk protein IgG and IgA: the association with milk-induced gastrointestinal symptoms in adults.

    Science.gov (United States)

    Anthoni, Sari; Savilahti, Erkki; Rautelin, Hilpi; Kolho, Kaija-Leena

    2009-10-21

    To study the association between serum levels of milk protein IgG and IgA antibodies and milk-related gastrointestinal symptoms in adults. Milk protein IgG and IgA antibodies were determined in serum samples of 400 subjects from five outpatient clinics in Southern Finland. Subjects were randomly selected from a total of 1900 adults undergoing laboratory investigations in primary care. All 400 participants had completed a questionnaire on abdominal symptoms and dairy consumption while waiting for the laboratory visit. The questionnaire covered the nature and frequency of gastrointestinal problems, the provoking food items, family history and allergies. Twelve serum samples were disqualified due to insufficient amount of sera. The levels of specific milk protein IgG and IgA were measured by using the ELISA technique. The association of the milk protein-specific antibody level was studied in relation to the milk-related gastrointestinal symptoms and dairy consumption. Subjects drinking milk (n = 265) had higher levels of milk protein IgG in their sera than non-milk drinkers (n = 123, P milk drinking (n = 119) consumed less milk but had higher milk protein IgG levels than those with no milk-related gastrointestinal symptoms (n = 198, P = 0.02). Among the symptomatic subjects, those reporting dyspeptic symptoms had lower milk protein IgG levels than non-dyspeptics (P milk drinking (P = 0.5). The association of high milk protein IgG levels with constipation was close to the level of statistical significance. Diarrhea had no association with milk protein IgG level (P = 0.5). With regard to minor symptoms, flatulence and bloating (P = 0.8), were not associated with milk protein IgG level. Milk protein IgA levels did not show any association with milk drinking or abdominal symptoms. The levels of milk protein IgA and IgG declined as the age of the subjects increased (P Milk protein IgG but not milk IgA seems to be associated with self-reported milk-induced gastrointestinal

  15. [Serum immunoglobulin IgG subclass distribution of antibody responses to pertussis toxin and filamentous hemagglutinin of Bordetella pertussis in patients with whooping cough].

    Science.gov (United States)

    Rastawicki, Waldemar; Smietańska, Karolina; Rokosz-Chudziak, Natalia; Jagielski, Marek

    2013-01-01

    The present study was aimed at determining the IgG subclass distribution against pertussis toxin (PT) and filamentous hemagglutinin (FHA) of Bordetella pertussis in patients with whooping cough. The total number of 222 serum samples obtained from patients suspected in clinical investigation for pertussis were tested separately by in-house ELISA for the presence of IgG antibodies to pertussis toxin and filamentous hemagglutinin. The percentage distribution of specific anti-PT and anti-FHA IgG subclass response was calculated only on the basis of group of sera confirmed in the present study as positive for total IgG antibodies (183 sera to PT antigen and 129 to FHA antigen). Paired serum specimens were obtained from 36 patients. Based on the results of determining the level of antibodies in the sera of 40 blood donors, the cut-off limit of serum antibodies for each subclass was set at arithmetic mean plus two standard deviations. Antibodies of IgG1 to pertussis toxin and filamentous hemagglutinin were diagnosed in 151 (82.5%) and 99 (76.7%), IgG2 in 72 (39.0%) and 50 (38.8%), IgG3 in 17 (9.3%) and 43 (33.3%), IgG4 in 55 (30.1%) and 53 (41.1%) serum samples, respectively. There were no significant differences in percentage of sera with IgG1, IgG2 and IgG3 in relation to age of the patients. However, the frequency of occurrence of IgG4 antibodies was highest in the group of the youngest children to the age of 6 years old (61.8% for PT and 68.0% for FHA), and decrease with age, reaching the minimum in the group of patients above 40 years old (13.2% and 4.2% for PT and FHA, respectively). We also found significantly higher frequency of IgG4 to PT and FHA antigens in men than in women. Statistically significant, essential changes in the pattern of IgG subclass during the course of infection were not found. In conclusion, this study showed that all four subclasses of IgG antibodies to pertussis toxin and filamentous hemagglutinin are produced during whooping cough.

  16. Cellphone-based hand-held microplate reader for point-of-care ELISA testing (Conference Presentation)

    Science.gov (United States)

    Berg, Brandon; Cortazar, Bingen; Tseng, Derek; Ozkan, Haydar; Feng, Steve; Wei, Qingshan; Chan, Raymond Y.; Burbano, Jordi; Farooqui, Qamar; Lewinski, Michael; Di Carlo, Dino; Garner, Omai B.; Ozcan, Aydogan

    2016-03-01

    Enzyme-linked immunosorbent assay (ELISA) in a microplate format has been a gold standard first-line clinical test for diagnosis of various diseases including infectious diseases. However, this technology requires a relatively large and expensive multi-well scanning spectrophotometer to read and quantify the signal from each well, hindering its implementation in resource-limited-settings. Here, we demonstrate a cost-effective and handheld smartphone-based colorimetric microplate reader for rapid digitization and quantification of immunoserology-related ELISA tests in a conventional 96-well plate format at the point of care (POC). This device consists of a bundle of 96 optical fibers to collect the transmitted light from each well of the microplate and direct all the transmission signals from the wells onto the camera of the mobile-phone. Captured images are then transmitted to a remote server through a custom-designed app, and both quantitative and qualitative diagnostic results are returned back to the user within ~1 minute per 96-well plate by using a machine learning algorithm. We tested this mobile-phone based micro-plate reader in a clinical microbiology lab using FDA-approved mumps IgG, measles IgG, and herpes simplex virus IgG (HSV-1 and HSV-2) ELISA tests on 1138 remnant patient samples (roughly 50% training and 50% testing), and achieved an overall accuracy of ~99% or higher for each ELISA test. This handheld and cost-effective platform could be immediately useful for large-scale vaccination monitoring in low-infrastructure settings, and also for other high-throughput disease screening applications at POC.

  17. Correlation of Fc(gamma)RIIa (CD32) Polymorphism and IgG Antibody Subclasses in Hemolytic Disease of Newborn.

    Science.gov (United States)

    Wu, QiangJu; Zhang, Yan; Liu, MengLi; Wang, Bo; Liu, Sheng; He, Chen

    2009-01-01

    ABO-HDN is a common disease of newborn in China and currently there is no satisfactory method to predict it in the antepartum period. It has been reported that Fc(gamma)RIIa (CD32) genotype is associated with both infectious diseases induced by bacteria and parasitemia. There is a relationship between IgG subclass and RH-HDN. To study the pathogenesis of ABO-HDN and to find reliable method to diagnose ABO-HDN, we investigated the polymorphism of Fc(gamma)RIIa (CD32) and distribution of IgG subclass in infants with ABO-HDN and their mothers by polymerase chain reaction or ELISA assay. We observed that the frequency of HH131 genotype is lower in infants with ABO-HDN than in controls (p < 0.01), while the frequency of HR131 genotype is higher in ABO-HDN infants than that in controls (p < 0.01). The genotype HR131 and concentrations of IgG1 and IgG3 are significantly correlated with ABO-HDN. Our results demonstrated, for the first time, that there is a correlation between ABO-HDN and CD32, and different IgG subclass distribution. Our study may contribute to the development of an early diagnostic method for HDN. Copyright 2009 S. Karger AG, Basel.

  18. Survey of malaria and anti-dengue virus IgG among febrile HIV-infected patients attending a tertiary hospital in Abuja, Nigeria.

    Science.gov (United States)

    Mustapha, Jelili Olaide; Emeribe, Anthony Uchenna; Nasir, Idris Abdullahi

    2017-01-01

    Dengue and malaria are infections, of great public health concern, especially in sub-Saharan Africa where the burden of HIV infection is high. This study was conducted to determine the seroprevalence of dengue virus IgG antibodies and dengue/malaria coinfection among febrile HIV-infected patients attending the University of Abuja Teaching Hospital, Gwagwalada, Abuja. In this cross-sectional study, blood samples from 178 consenting HIV-infected patients receiving antiretroviral therapy were collected and tested for plasmodiasis and anti-Dengue virus IgG using malaria microscopy and ELISA, respectively. Interviewer-based questionnaires were used to assess subjects' sociodemographic variables and dengue risk factors. Of the 178 screened participants, 44.4% were seropositive for dengue virus IgG antibody, whereas 29.2% were positive for Plasmodium falciparum. About 44.2% were positive for both dengue virus and P. falciparum . There was a statistical association between anti-dengue IgG and occupation ( p =0.03) but not with age, residential area, educational level and patients' gender ( p >0.05). Seroprevalence of anti-dengue specific IgG was relatively higher in participants who adopted protective measures. There was a statistical association between seroprevalence of anti-dengue IgG and adoption of preventive measures ( p <0.05). The high prevalence of malaria and dengue virus IgG indicates the need to strengthen vector control and dengue surveillance programs.

  19. Dengue Virus NS1 Protein as a Diagnostic Marker: Commercially Available ELISA and Comparison to qRT-PCR and Serological Diagnostic Assays Currently Used by the State of Florida

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    Jason H. Ambrose

    2017-01-01

    Full Text Available Background. The proper management of patients infected with dengue virus requires early detection. Here, real-time molecular assays have proven useful but have limitations, whereas ELISAs that detect antibodies are still favored but results are obtained too late to be of clinical value. The production of DENV NS1 peaks early during infection and its detection can combine the advantages of both diagnostic approaches. Methods. This study compared assays currently used for detecting DENV infection at the Florida Department of Health including anti-DENV IgM and IgG ELISAs as well as qRT-PCR, against a commercially available DENV NS1 ELISA. These comparisons were made among a group of 21 human sera. Results. Nine of 14 (64.3% DENV qRT-PCR+ samples were also DENV NS1+. Interestingly, the 5 NS1− samples that were qRT-PCR+ were additionally IgM− and IgG+ suggesting a nonprimary infection. Compared to qRT-PCR, the NS1 assay had a sensitivity of 64.3%, specificity 100%, PPV of 100%, and NPV of 58.3%. Conclusions. The NS1 ELISA performed as expected in known DENV qRT-PCR+ samples; however negative NS1 results for qRT-PCR+ and IgG+ sera seemingly reduced the usefulness of the NS1 ELISA for nonprimary cases. We therefore conclude that diagnosis obtained via DENV NS1 ELISA deserves further investigation.

  20. Clinical significance of pretreatment FDG PET/CT IN MOBG-avid pediatric neuroblastoma

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    Kang, Seo Young; Kim, Yong Il; Cheon, Gi Jeong; Kang, Keon Wook; Chung, June Key; Lee, Dong Soo; Kang, Hyoung Jin; Shin, Hee Young [Seoul National University Hospital, Seoul (Korea, Republic of); Kim, E. Edmund [Seoul National University, Seoul (Korea, Republic of); Rahim, Muhammad Kashif [Nishtar Medical College and Hospital, Multan (Pakistan)

    2017-06-15

    {sup 18}F-fluorodeoxyglucose-positron emission tomography (FDG-PET) imaging is well known to have clinical significance in the initial staging and response evaluation of the many kinds of neoplasms. However, its role in the pediatric neuroblastoma is not clearly defined. In the present study, the clinical significance of FDG-PET/computed tomography (CT) in 123I- or 131I-metaiodobenzylguanidine (MIBG)-avid pediatric neuroblastoma was investigated. Twenty patients with neuroblastoma who undertook pretreatment FDG PET/CT at our institute between 2008 and 2015 and showed MIBG avidity were retrospectively enrolled in the present study. Clinical information—including histopathology, and serum markers—and several PET parameters—including SUVmax of the primary lesion (Psuv), target-to-background ratio (TBR), metabolic tumor volume (MTV), and coefficient of variation (CV)—were analyzed. The prognostic effect of PET parameters was evaluated in terms of progression-free survival (PFS). Total 20 patients (4.5 ± 3.5 years) were divided as two groups by disease progression. Six patients (30.0 %) experienced disease progression and one patient (5.0 %) died during follow-up period. There were not statistically significant in age, stage, MYCN status, primary tumor size, serum lactate dehydrogenase (LDH), neuron-specific enolase (NSE), and ferritin level between two groups with progression or no progression. However, Psuv (p = 0.017), TBR (p = 0.09), MTV (p = 0.02), and CV (p = 0.036) showed significant differences between two groups. In univariate analysis, PFS was significantly associated with Psuv (p = 0.021) and TBR (p = 0.023). FDG-PET parameters were significantly related with progression of neuroblastoma. FDG-PET/CT may have the potential as a valuable modality for evaluating prognosis in the patients with MIBG-avid pediatric neuroblastoma.

  1. Determination of Seroprevalence of Borrelia burgdorferi IgG in Adult Population Living in Trabzon.

    Science.gov (United States)

    Cora, Merve; Kaklıkkaya, Neşe; Topbaş, Murat; Çan, Gamze; Yavuzyılmaz, Asuman; Tosun, İlknur; Aydın, Faruk

    2017-01-01

    Lyme borreliosis is a tick-borne, multi-systemic infectious disease that is thought to be wide spread in Turkey even though studies on its seroprevalence are limited. To determine the seroprevalence of Lyme borreliosis in part of north-eastern Tur-key (in the city of Trabzon), and to identify possible relationships between seropositivity and various factors such as location, gender, age group, occupation, income, and educational level. Retrospective cross-sectional study. A total of 884 blood samples collected from provincial and district health centers serving a population of about 800,000 were included in this study. ELISA was used to determine the anti-Borrelia IgG antibody levels in the samples. Samples that yielded positive results by ELISA were further subjected to western blot (WB). IgG antibodies were found in 128 samples (14.5%). Statistical analysis revealed significant differences between age groups and educational levels in terms of the incidence of seropositivity, whereas location, gender, occupational group and income level had no effect (p<0.001, p<0.001, p=0.948, p=0.645, p=0.131, p=0.080 respectively). The risk of contracting Lyme borreliosis in Trabzon is high, and necessary measures need to be taken to avoid the spread of disease.

  2. Determination of Seroprevalence of Borrelia burgdorferi IgG in Adult Population Living in Trabzon

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    Merve Cora

    2017-02-01

    Full Text Available Background: Lyme borreliosis is a tick-borne, multi-systemic infectious disease that is thought to be wide spread in Turkey even though studies on its seroprevalence are limited. Aims: To determine the seroprevalence of Lyme borreliosis in part of north-eastern Tur-key (in the city of Trabzon, and to identify possible relationships between seropositivity and various factors such as location, gender, age group, occupation, income, and educational level. Study Design: Retrospective cross-sectional study. Methods: A total of 884 blood samples collected from provincial and district health centers serving a population of about 800,000 were included in this study. ELISA was used to determine the anti-Borrelia IgG antibody levels in the samples. Samples that yielded positive results by ELISA were further subjected to western blot (WB. Results: IgG antibodies were found in 128 samples (14.5%. Statistical analysis revealed significant differences between age groups and educational levels in terms of the incidence of seropositivity, whereas location, gender, occupational group and income level had no effect (p<0.001, p<0.001, p=0.948, p=0.645, p=0.131, p=0.080 respectively. Conclusions: The risk of contracting Lyme borreliosis in Trabzon is high, and necessary measures need to be taken to avoid the spread of disease

  3. The modification and evaluation of an ELISA test for the surveillance of Mycobacterium avium subsp. paratuberculosis infection in wild ruminants

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    Pruvot Mathieu

    2013-01-01

    Full Text Available Abstract Background Enzyme-linked immunosorbent assay (ELISA is often used to test wildlife samples for Mycobacterium avium subsp. paratuberculosis (MAP infection. However, commercially available kits are only validated for use with domestic ruminant species. A literature review was performed to document the current use of MAP serum ELISA in wild and semi-domestic ruminants. We then modified and evaluated a commercial ELISA kit (IDEXX Mycobacterium paratuberculosis Antibody Test Kit for use with species for which it was not originally developed: elk (Cervus elaphus, bison (Bison bison and caribou (Rangifer tarandus. We tested the affinity of different conjugates for immunoglobulin G (IgG isolated from these species, performed checkerboard tests to determine the optimal dilutions of samples and conjugates, and established cut-off values using two different methods: a Receiver Operational Curve on a panel of known samples for elk, and an alternate method involving a panel of unknown serum samples for the three species. Results We found that the anti-bovine conjugate included in the IDEXX ELISA kit has limited affinity for elk, bison, and caribou IgG. Protein G showed good affinity for IgG of all three species, while anti-deer conjugate also bound elk and caribou IgG. Using Protein G with elk serum, a cut-off sample-to-positive (S/P value of 0.22 was selected, resulting in a sensitivity and specificity of 73% and 90%, respectively, whereas, using an anti-deer conjugate with elk serum, an S/P cut-off value of 0.29 gave a sensitivity of 68%, with 100% specificity. Cut-off values for bison and caribou using the Protein G conjugate were 0.17 and 0.25 respectively. Conclusions Due to incomplete reporting and a lack of test validation, it is difficult to critically appraise results of many sero-surveys that have previously been done for MAP in wildlife. Commercial ELISA kits may have limited or no capacity to detect antibodies from species other than for

  4. Specific IgA and IgG antibodies in paired serum and breast milk samples in human strongyloidiasis.

    Science.gov (United States)

    Mota-Ferreira, Daniela M L; Gonçalves-Pires, Maria do Rosário F; Júnior, Alvaro Ferreira; Sopelete, Mônica C; Abdallah, Vânia O S; Costa-Cruz, Julia M

    2009-02-01

    Strongyloidiasis, caused by the nematode Strongyloides stercoralis, is one of the major worldwide parasitic infections in humans. Breastfeeding may offer a potential protection against this infection. Feces, serum and milk samples were obtained from 90 lactating women from Clinical Hospital of Universidade Federal de Uberlândia, Brazil. The fecal samples were collected for parasitological diagnosis and the serum and milk samples were examined for specific S. stercoralis IgA and IgG antibodies using the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Fecal examination showed that the rate of prevalence of S. stercoralis infection in the lactating women was 4.4%. IFAT manifested a 16.7% positivity rate for specific IgA antibody in serum and a 28.9% rate in milk samples; specific IgG was 41.1% in serum and 25.5% in milk samples. According to ELISA the positivity rate for specific IgA antibody was 21.1% in serum and 42.2% in milk samples; specific IgG was 40% in serum and 18.9% in milk samples. In serum samples, these immunological tests showed a concurrence of 91.1% and 94.4%, respectively, in detecting specific IgA and IgG antibodies. In milk samples, they showed a concurrence of 70% and 78.9%, respectively, in detecting specific IgA and IgG antibodies. There was a statistically significant difference between concordant and discordant results of immunological tests (Pantibodies in serum and in milk samples reconfirming prior studies that the serological method is a complement to the direct diagnosis of the parasite, and suggesting that immunological methods using milk samples can also be helpful. Furthermore, in endemic areas, infants may acquire antibodies to S. stercoralis from breast milk, possibly, contributing to the enhancement of specific mucosal immunity against this parasite.

  5. Elisa Studio showroom = Elisa Studio showroom / Jan Joonas Graps ; kommenteerinud Tiina Kuusisto

    Index Scriptorium Estoniae

    Graps, Jan Joonas, 1972-

    2015-01-01

    Elisa Studio showroom Helsingis Lasiplatsi, Mannerheimintie 22-24. Sisekujunduse autorid Jan Joonas Graps, Ken-Kristjan Ruut, Anne Määrmann (JanKen Wisespace). Lühidalt loovstuudiost JanKen Wisespace

  6. Saliva and sera IgA and IgG in Egyptian Giardia-infected children.

    Science.gov (United States)

    El-Gebaly, Naglaa Saad M; Halawa, Eman Fawzy; Moussa, Hanaa M Ezzat; Rabia, Ibrahim; Abu-Zekry, Maha

    2012-08-01

    Giardiasis is a gastrointestinal infection of wide distribution that is more prevalent in childhood. Easy and rapid diagnosis of giardiasis is essential for reduction of this infection. This cross-sectional study included 62 children in which collection of saliva, stool and serum samples was performed. An enzyme-linked immunosorbent assay (ELISA) technique was evaluated to detect IgA and IgG responses in both saliva and serum samples. Twenty-two children were positive for Giardia duodenalis infection by direct examination of faecal specimens, 20 non-infected and 20 infected with other parasites. Salivary and serum IgA and IgG responses against G. duodenalis infection were significantly higher in Giardia parasitized than non-Giardia parasitized children (p < 0.001). This concludes that specific salivary IgA may serve as a diagnostic tool and specific salivary IgG as a screening tool in monitoring the exposure of various populations to Giardia duodenalis. The advantage of salivary assays over serum immunoglobulin assay is being easy and non-invasive in sampling technique which is important especially for young children.

  7. Altered glycosylation of complexed native IgG molecules is associated with disease activity of systemic lupus erythematosus.

    Science.gov (United States)

    Sjöwall, C; Zapf, J; von Löhneysen, S; Magorivska, I; Biermann, M; Janko, C; Winkler, S; Bilyy, R; Schett, G; Herrmann, M; Muñoz, L E

    2015-05-01

    In addition to the redundancy of the receptors for the Fc portion of immunoglobulins, glycans result in potential ligands for a plethora of lectin receptors found in immune effector cells. Here we analysed the exposure of glycans containing fucosyl residues and the fucosylated tri-mannose N-type core by complexed native IgG in longitudinal serum samples of well-characterized patients with systemic lupus erythematosus. Consecutive serum samples of a cohort of 15 patients with systemic lupus erythematosus during periods of increased disease activity and remission were analysed. All patients fulfilled the 1982 American College of Rheumatology classification criteria. Sera of 15 sex- and age-matched normal healthy blood donors served as controls. The levels and type of glycosylation of complexed random IgG was measured with lectin enzyme-immunosorbent assays. After specifically gathering IgG complexes from sera, biotinylated lectins Aleuria aurantia lectin and Lens culinaris agglutinin were employed to detect IgG-associated fucosyl residues and the fucosylated tri-mannose N-glycan core, respectively. In sandwich-ELISAs, IgG-associated IgM, IgA, C1q, C3c and C-reactive protein (CRP) were detected as candidates for IgG immune complex constituents. We studied associations of the glycan of complexed IgG and disease activity according to the physician's global assessment of disease activity and the systemic lupus erythematosus disease activity index 2000 documented at the moment of blood taking. Our results showed significantly higher levels of Aleuria aurantia lectin and Lens culinaris agglutinin binding sites exposed on IgG complexes of patients with systemic lupus erythematosus than on those of normal healthy blood donors. Disease activity in systemic lupus erythematosus correlated with higher exposure of Aleuria aurantia lectin-reactive fucosyl residues by immobilized IgG complexes. Top levels of Aleuria aurantia lectin-reactivity were found in samples taken during the

  8. A novel antigen capture ELISA for the specific detection of IgG antibodies to elephant endotheliotropic herpes virus

    NARCIS (Netherlands)

    P. van den Doel (Petra); V.R. Prieto (Víctor Rodríguez); S.E. Van Rossum-Fikkert (Sarah E.); W. Schaftenaar (Willem); E. Latimer (Erin); L. Howard (Lauren); S. Chapman (Sarah); N. Masters (Nic); A.D.M.E. Osterhaus (Albert); P.D. Ling (Paul D.); A. Dastjerdi (Akbar); B.E.E. Martina (Byron)

    2015-01-01

    textabstractBackground: Elephants are classified as critically endangered animals by the International Union for Conservation of Species (IUCN). Elephant endotheliotropic herpesvirus (EEHV) poses a large threat to breeding programs of captive Asian elephants by causing fatal haemorrhagic disease.

  9. Reliability of an ELISA test for diagnosing oestrosis in Iberian ibex.

    Science.gov (United States)

    Arias, María Sol; Moreno, Virginia; Sarasa, Mathieu; Paz-Silva, Adolfo; Sánchez-Andrade, Rita; Morrondo, Patrocinio; Díez-Baños, Pablo; Granados, José E; Sánchez, Antonio; Pérez, Jesús M

    2014-04-01

    Oestrosis is one of the most prevalent parasitosis affecting the Iberian ibex, Capra pyrenaica . To date, both the diagnosis of oestrosis and the determination of the intensity of parasitism require the use of invasive methods (necropsy), which necessarily limit research possibilities. We analyzed the immune humoral response (IgM and IgG) against Oestrus ovis L. excretory/secretory larval antigens in 32 sera taken from Iberian ibex from the Sierra Nevada Natural Space (southern Spain). Three antigens were collected: L1OES (from L1 larvae), L2OES (L2), and L3OES (L3). Necropsy was considered as the gold standard. The percentage of ibexes harboring Oestrus spp. larvae was 88%, the mean intensity of parasitism being 16.96 ± 14.96 larvae per parasitized host (range: 2-52). In our sample, first-instar larvae (L1) were found in 9% of ibexes, while 69% of hosts carried L2 larvae and 88% L3 larvae. Positive correlations between L1 and L2 numbers, and between L2 and L3 numbers were detected. The best results with the immunoenzymatic assay were obtained using IgG antibodies against the L1OES antigens (specificity = 89%; sensitivity = 100%; positive predictive value = 100%; negative predictive value = 57%). The IgG seroprevalence against L1OES was 78%. Thus, the analysis of IgG antibodies against antigens collected from L1 O. ovis larvae would seem to be a noninvasive method for reliably diagnosing oestrosis in naturally infested Iberian ibex. Nevertheless, additional immunological and methodological advances are still required because false positive and false negative results still represent a non-negligible part of the results of the ELISA tests.

  10. Serological follow-up of patients with paracoccidioidomycosis treated with itraconazole using Dot-blot, ELISA and Western-blot

    Directory of Open Access Journals (Sweden)

    Rosana MARTINS

    1997-09-01

    Full Text Available Twenty-seven mycologically proven cases of paracoccidioidomycosis (PCM were treated with itraconazole (100-200 mg/day in month 1 and 100 mg/day until month 6-8 and evaluated clinically and serologically, up to 3.5 years post-therapy, using Dot-blot and ELISA for measuring the titers of IgG, IgA and IgM anti-P. brasiliensis antibodies and Western-blot for determining IgG, IgA and IgM antibodies against the antigen components of the fungus. Before treatment, 81.5% (Dot-blot and 84% (ELISA of the patients presented elevated IgG anti-P. brasiliensis antibody titers which dropped slightly with treatment. On the other hand, the percentages of pre-treatment high-titered sera for IgA and IgM anti-P.brasiliensis were lower (5l.9% and 5l.8%: Dot-blot; 16.5 and 36%: ELISA, respectively but the titers tended to become negative more frequently with treatment. Prior to treatment, the percentages of positivity for IgG, IgA and IgM anti-P.brasiliensis antibodies in Western-blot were 96%, 20.8% and 41.6%, respectively. Antigens with molecular weights varying from 16-78 kDa, from 21-76 kDa and from 27-78 kDa were reactive for IgG, IgA and IgM antibodies, respectively. The most frequently reactive antigenic components had molecular weights of 27, 33 and 43 kDa for IgG, and 70 for IgA and IgM antibodies. During the period of study, the patients responded well to treatment. The present data confirm the diversity and complexity of the humoral response in PCM, and the importance of utilizing different serological tests to detect IgG, IgA and IgM anti-P. brasiliensis antibodiesVinte e sete pacientes portadores de paracoccidioidomicose (PCM foram tratados com itraconazole (100-200 mg/dia no primeiro mês e 100 mg/dia até 6-8 meses e avaliados sob o ponto de vista clínico e sorológico, até 3 e meio anos após o início do tratamento, utilizando-se os testes de Dot-blot e ELISA para medir os títulos de anticorpos IgG, IgA e IgM anti-P. brasiliensis, e Western

  11. IgG abnormality in narcolepsy and idiopathic hypersomnia.

    Directory of Open Access Journals (Sweden)

    Susumu Tanaka

    Full Text Available BACKGROUND: A close association between narcolepsy and the Human Leukocyte Antigen (HLA-DQB1*0602 allele suggests the involvement of the immune system, or possibly an autoimmune process. We investigated serum IgG levels in narcolepsy. METHODOLOGY/PRINCIPAL FINDINGS: We measured the serum total IgG levels in 159 Japanese narcolepsy-cataplexy patients positive for the HLA-DQB1*0602 allele, 28 idiopathic hypersomnia patients with long sleep time, and 123 healthy controls (the HLA-DQB1*0602 allele present in 45 subjects. The serum levels of each IgG subclass were subsequently measured. The distribution of serum IgG was significantly different among healthy controls negative for the HLA-DQB1*0602 allele (11.66+/-3.55 mg/ml, healthy controls positive for the HLA-DQB1*0602 allele (11.45+/-3.43, narcolepsy patients (9.67+/-3.38, and idiopathic hypersomnia patients (13.81+/-3.80. None of the following clinical variables, age, disease duration, Epworth Sleepiness Scale, smoking habit and BMI at the time of blood sampling, were associated with IgG levels in narcolepsy or idiopathic hypersomnia. Furthermore we found the decrease in IgG1 and IgG2 levels, stable expression of IgG3, and the increase in the proportion of IgG4 in narcolepsy patients with abnormally low IgG levels. The increase in the proportion of IgG4 levels was also found in narcolepsy patients with normal serum total IgG levels. Idiopathic hypersomnia patients showed a different pattern of IgG subclass distribution with high IgG3 and IgG4 level, low IgG2 level, and IgG1/IgG2 imbalance. CONCLUSIONS/SIGNIFICANCE: Our study is the first to determine IgG abnormalities in narcolepsy and idiopathic hypersomnia by measuring the serum IgG levels in a large number of hypersomnia patients. The observed IgG abnormalities indicate humoral immune alterations in narcolepsy and idiopathic hypersomnia. Different IgG profiles suggest immunological differences between narcolepsy and idiopathic hypersomnia.

  12. First preclinical evaluation of mono-[123I]iodohypericin as a necrosis-avid tracer agent.

    Science.gov (United States)

    Ni, Yicheng; Huyghe, Dieter; Verbeke, Kristin; de Witte, Peter A; Nuyts, Johan; Mortelmans, Luc; Chen, Feng; Marchal, Guy; Verbruggen, Alfons M; Bormans, Guy M

    2006-05-01

    We have labelled hypericin, a polyphenolic polycyclic quinone found in St. John's wort (Hypericum perforatum), with( 123)I and evaluated mono-[(123)I]iodohypericin (MIH) as a potential necrosis-avid diagnostic tracer agent. MIH was prepared by an electrophilic radioiodination method. The new tracer agent was evaluated in animal models of liver infarction in the rat and heart infarction in the rabbit using single-photon emission computed tomography (SPECT), triphenyltetrazolium chloride (TTC) histochemical staining, serial sectional autoradiography and microscopy, and radioactivity counting techniques. Using in vivo SPECT imaging, hepatic and cardiac infarctions were persistently visualised as well-defined hot spots over 48 h. Preferential uptake of the tracer agent in necrotic tissue was confirmed by perfect match of images from post-mortem TTC staining, autoradiography (ARX) and histology. Radioactivity concentration in infarcted tissues was over 10 times (liver; 3.51% ID/g in necrotic tissue vs 0.38% ID/g in normal tissue at 60 h p.i.) and over 6 times (myocardium; 0.36% ID/g in necrotic tissue vs 0.054% ID/g in normal tissue; ratios up to 18 for selected parts on ARX images) higher than in normal tissues. The results suggest that hypericin derivatives may serve as powerful necrosis-avid diagnostic agents for assessment of tissue viability.

  13. Vaccination for birch pollen allergy. Induction of affinity-matured or blocking IgG antibodies does not account for the reduced binding of IgE to Bet v 1

    DEFF Research Database (Denmark)

    Svenson, Morten; Jacobi, Henrik H; Bødtger, Uffe

    2003-01-01

    (Betula verrucosa) (Bet v 1), the major allergen in birch pollen, to serum IgG and IgE, separately and in competition. Sera from six birch pollen-allergic patients were obtained before and after 5 years of SAV, and binding was assessed with 125I-Bet v 1. Before SAV, IgG bound more than eight times...... the amount of Bet v 1 compared with IgE, and together they accounted for more than 85% of the serum binding capacity. While SAV induced minimal changes in IgE binding, the IgG binding capacities increased 6-32 times. In contrast, the binding avidities (K(d) 28-40pM) changed less than 20%, pre- and post......-SAV IgG provided similar inhibition of Bet v 1 binding to IgE at equimolar levels, and cross inhibition studies between IgG and IgE showed low inter-individual differences. Following SAV, all sera reduced Bet v 1 binding to CD23(+) cells, correlating with reduced binding of Bet v 1 to IgE (P...

  14. Vaccination for birch pollen allergy. Induction of affinity-matured or blocking IgG antibodies does not account for the reduced binding of IgE to Bet v 1

    DEFF Research Database (Denmark)

    Svenson, Morten; Jacobi, Henrik H; Bødtger, Uffe

    2003-01-01

    (Betula verrucosa) (Bet v 1), the major allergen in birch pollen, to serum IgG and IgE, separately and in competition. Sera from six birch pollen-allergic patients were obtained before and after 5 years of SAV, and binding was assessed with 125I-Bet v 1. Before SAV, IgG bound more than eight times...... the amount of Bet v 1 compared with IgE, and together they accounted for more than 85% of the serum binding capacity. While SAV induced minimal changes in IgE binding, the IgG binding capacities increased 6-32 times. In contrast, the binding avidities (K(d) 28-40pM) changed less than 20%, pre- and post......-SAV IgG provided similar inhibition of Bet v 1 binding to IgE at equimolar levels, and cross inhibition studies between IgG and IgE showed low inter-individual differences. Following SAV, all sera reduced Bet v 1 binding to CD23(+) cells, correlating with reduced binding of Bet v 1 to IgE (P

  15. Hyperglucagonaemia analysed by glucagon sandwich ELISA

    DEFF Research Database (Denmark)

    Albrechtsen, Nicolai Jacob Wewer; Hartmann, Bolette; Veedfald, Simon

    2014-01-01

    after RYGB were from a study by Bojsen-Møller et al (trial registration number NCT 01202526). Samples from vagotomised and control individuals were from a study by Plamboeck et al (NCT01176890). Samples from ESRD patients were from a study by Idorn et al (NCT01327378)....... the extent to which the hyperglucagonaemia measured in clinical samples was caused by authentic glucagon. METHODS: We examined the performance of three commercial glucagon 'sandwich' ELISAs. The ELISA with the best overall performance was selected to compare glucagon measurements in clinical samples...... with an established glucagon RIA. RESULTS: The first assay performed poorly: there was high cross-reactivity with glicentin (22%) and a lack of sensitivity for glucagon. The second and third assays showed minor cross-reactivity (1-5%) with oxyntomodulin and glicentin; however, the second assay had insufficient...

  16. The feasibility of harmonizing gluten ELISA measurements.

    Science.gov (United States)

    Rzychon, Malgorzata; Brohée, Marcel; Cordeiro, Fernando; Haraszi, Reka; Ulberth, Franz; O'Connor, Gavin

    2017-11-01

    Many publications have highlighted that routine ELISA methods do not give rise to equivalent gluten content measurement results. In this study, we assess this variation between results and its likely impact on the enforcement of the EU gluten-free legislation. This study systematically examines the feasibility of harmonizing gluten ELISA assays by the introduction of: a common extraction procedure; a common calibrator, such as a pure gluten extract and an incurred matrix material. The comparability of measurements is limited by a weak correlation between kit results caused by differences in the selectivity of the methods. This lack of correlation produces bias that cannot be corrected by using reference materials alone. The use of a common calibrator reduced the between-assay variability to some extent, but variation due to differences in selectivity of the assays was unaffected. Consensus on robust markers and their conversion to "gluten content" are required. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  17. Optimization of ELISA Conditions to Quantify Colorectal Cancer Antigen-Antibody Complex Protein (GA733-FcK) Expressed in Transgenic Plant

    OpenAIRE

    Ahn, Junsik; Lee, Kyung Jin; Ko, Kisung

    2014-01-01

    The purpose of this study is to optimize ELISA conditions to quantify the colorectal cancer antigen GA733 linked to the Fc antibody fragment fused to KDEL, an ER retention motif (GA733-FcK) expressed in transgenic plant. Variable conditions of capture antibody, blocking buffer, and detection antibody for ELISA were optimized with application of leaf extracts from transgenic plant expressing GA733-FcK. In detection antibody, anti-EpCAM/CD362 IgG recognizing the GA733 did not detect any GA733-F...

  18. Detection of IgM antibodies against canine distemper virus in dog and mink sera employing enzyme-linked immunosorbent assay (ELISA).

    Science.gov (United States)

    Blixenkrone-Møller, M; Pedersen, I R; Appel, M J; Griot, C

    1991-01-01

    An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against canine distemper virus (CDV) in canine and mink serum is described. The diagnostic potential of this technique was evaluated by analyzing sera from natural or experimental infections in dog and mink and negative control sera. These results were compared with results obtained in the developed CDV IgG ELISA and in the virus neutralization test. The IgM test, which requires only a single serum specimen, is a useful method for diagnosing current or recent CDV infections in dog and mink.

  19. Pregnancy does not affect HIV incidence test results obtained using the BED capture enzyme immunoassay or an antibody avidity assay.

    Directory of Open Access Journals (Sweden)

    Oliver Laeyendecker

    2010-10-01

    Full Text Available Accurate incidence estimates are needed for surveillance of the HIV epidemic. HIV surveillance occurs at maternal-child health clinics, but it is not known if pregnancy affects HIV incidence testing.We used the BED capture immunoassay (BED and an antibody avidity assay to test longitudinal samples from 51 HIV-infected Ugandan women infected with subtype A, C, D and intersubtype recombinant HIV who were enrolled in the HIVNET 012 trial (37 baseline samples collected near the time of delivery and 135 follow-up samples collected 3, 4 or 5 years later. Nineteen of 51 women were also pregnant at the time of one or more of the follow-up visits. The BED assay was performed according to the manufacturer's instructions. The avidity assay was performed using a Genetic Systems HIV-1/HIV-2 + O EIA using 0.1M diethylamine as the chaotropic agent.During the HIVNET 012 follow-up study, there was no difference in normalized optical density values (OD-n obtained with the BED assay or in the avidity test results (% when women were pregnant (n = 20 results compared to those obtained when women were not pregnant (n = 115; for BED: p = 0.9, generalized estimating equations model; for avidity: p = 0.7, Wilcoxon rank sum. In addition, BED and avidity results were almost exactly the same in longitudinal samples from the 18 women who were pregnant at only one study visit during the follow-up study (p = 0.6, paired t-test.These results from 51 Ugandan women suggest that any changes in the antibody response to HIV infection that occur during pregnancy are not sufficient to alter results obtained with the BED and avidity assays. Confirmation with larger studies and with other HIV subtypes is needed.

  20. Comparison of two commercial vaccines against visceral leishmaniasis in dogs from endemic areas: IgG, and subclasses, parasitism, and parasite transmission by xenodiagnosis.

    Science.gov (United States)

    Fernandes, Consuelo Barreto; Junior, Jairo Torres Magalhães; de Jesus, Clauceane; Souza, Bárbara Maria Paraná da Silva; Larangeira, Daniela Farias; Fraga, Deborah Bittencourt Mothé; Tavares Veras, Patricia Sampaio; Barrouin-Melo, Stella Maria

    2014-03-05

    The incidence of zoonotic canine visceral leishmaniasis (CVL) would decrease if dogs were effectively vaccinated; however, additional data on the efficacy of canine vaccines are required for their approved preventative use. To prospectively evaluate vaccination outcomes using two products commercially available in Brazil, with respect to adverse reactions (reactogenicity), humoral response, disease signs, parasitism, and parasite infectiousness in naturally exposed pet dogs in an endemic area of visceral leishmaniasis (VL). From 2010 to 2012, healthy dogs were vaccinated with Leishmune(®) (50 animals) or Leish-Tec(®) (50 animals). Each dog was examined to identify clinical signs during peri- and post-vaccination procedures every 2 months for 11 months to identify the presence of parasites or parasite DNA in splenic samples using culturing or PCR, respectively. Levels of anti-Leishmania IgG, IgG1, and IgG2 were quantified in sera by ELISA and infectiousness was assessed by xenodiagnosis. Adverse effects occurred in 2.2% (1/45) and 13.0% (6/46) of the animals in the Leishmune(®) and Leish-Tec(®) groups, respectively. IgG levels peaked on the 21st day following the first dose of Leishmune(®) and on the 21st day after the second dose of Leish-Tec(®). The final seropositivity rate for IgG was 32.5% (13/40) and 30.9% (13/42) in the Leishmune(®) and Leish-Tec(®) groups, respectively. The Leishmune(®) group presented higher levels of IgG1 and IgG2 compared to the Leish-Tec(®) group (p<0.001), and ELISA reactivity in both vaccinated groups was significantly lower (p<0.001) than in infected positive control dogs. Parasitism was observed in 12.2% (5/41) of the Leishmune(®) group, and 7.9% (3/38) of the Leish-Tec(®) group, with xenodiagnostic transmission rates of Leishmania to Lutzomyia longipalpis of 5.1% (2/39), and 5.4% (2/37), respectively. No significant differences were observed in dogs vaccinated with Leishmune(®) or Leish-Tec(®), with respect to LVC

  1. Paracoccidioides brasiliensis and P. lutzii antigens elicit different serum IgG responses in chronic paracoccidioidomycosis.

    Science.gov (United States)

    Lenhard-Vidal, A; Assolini, J P; Ono, M A; Bredt, C S O; Sano, A; Itano, E N

    2013-12-01

    Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis (S1, PS2, and PS3) and by the new species, P. lutzii. Considering that genetic differences in the Paracoccidioides genus could elicit distinct immune responses by the host, current research investigated serum IgG levels to antigens from P. brasiliensis B339 (S1), P. brasiliensis LDR3 (PS2), and atypical strain LDR2 (P. lutzii), in patients with chronic PCM from the northern and west regions of Paraná, Brazil (n = 35). Cell-free antigen (CFA) and high molecular mass fraction (hMM) were produced from each strain. Samples were analyzed by ELISA and immunodiffusion (ID). ELISA positivity using CFA: B339-100 %, LDR3-83 %, and LDR2-74 %. Response to CFA from B339 was more intense (p < 0.05), while there was no difference between LDR3-LDR2. IgG anti-hMM was higher for antigens from B339 or LDR3, when compared with LDR2 (p < 0.05). There was a positive correlation for each strain between CFA-hMM and for hMM between B339-LDR3 and LDR3-LDR2. ID positivity with CFA: B339-63 %, LDR3-66 %, and LDR2-60 %. We conclude that the intensity of reaction of the patients' sera varies with the strain used; hMM influences tests that use CFA, independently of strain; using ID, positive rates were very similar, but there was a large number of false negative results; ELISA tests using antigens from P. brasiliensis S1 were able to detect a larger number of patients than PS2 and P. lutzii (which had a considerable number of false negative results), and therefore, its use may be more appropriate in this region of Brazil.

  2. Inhibition of complement activation by IgG4 antibodies

    NARCIS (Netherlands)

    van der Zee, J. S.; van Swieten, P.; Aalberse, R. C.

    1986-01-01

    Prolonged exposure to antigens may result in high IgG4 antibody titres as was shown in a previous paper (Aalberse et al., 1983b). In novice bee keepers, a shift in the IgG1/IgG4 ratio of the response against phospholipase-A (PLA; a major component of bee venom) occurred. This resulted in an

  3. "Quasi-symmetric" FDG avidity of bilateral salivary glands in Mikulicz disease yielding false-positive staging in lung cancer.

    Science.gov (United States)

    Lin, Li-Fan; Shen, Daniel Hy; Tsai, Wen-Chiuan; Hou, Cheng-Han; Cheng, Cheng-Yi; Chen, Ching-Yuan

    2012-11-01

    A 77-year-old man with lung cancer underwent fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT). Cervical FDG uptake of variable intensity was detected corresponding to enlarged masses in the bilateral parotid and left submandibular regions. We suspected metastases from primary lung cancer or coexisting salivary gland neoplasms. Histopathological examination of the FDG-avid parotid mass revealed Mikulicz disease, an uncommon autoimmune disease involving the salivary glands. We present this case of Mikulicz disease with "quasi-symmetric" FDG avidity in the major salivary glands. We believe that salivary FDG uptake can lead to erroneous interpretation of tumor staging using PET.

  4. A Unique Report: Development of Super Anti-Human IgG Monoclone with Optical Density Over Than 3

    Directory of Open Access Journals (Sweden)

    Leili Aghebati Maleki

    2013-08-01

    Full Text Available Purpose: Monoclonal antibodies and related conjugates are key reagents used in biomedical researches as well as, in treatment, purification and diagnosis of infectious and non- infectious diseases. Methods: Balb/c mice were immunized with purified human IgG. Spleen cells of the most immune mouse were fused with SP2/0 in the presence of Poly Ethylene Glycol (PEG. Supernatant of hybridoma cells was screened for detection of antibody by ELISA. Then, the sample was assessed for cross-reactivity with IgM & IgA by ELISA and confirmed by immunoblotting. The subclasses of the selected mAbs were determined. The best clone was injected intraperitoneally to some pristane-injected mice. Anti-IgG mAb was purified from the animals' ascitic fluid by Ion exchange chromatography and then, mAb was conjugated with HRP. Results: In the present study, over than 50 clones were obtained that 1 clone had optical density over than 3. We named this clone as supermonoclone which was selected for limiting dilution. The result of the immunoblotting, showed sharp band in IgG position and did not show any band in IgM&IgA position. Conclusion: Based on the findings of this study, the conjugated monoclonal antibody could have application in diagnosis of infectious diseases like Toxoplasmosis, Rubella and IgG class of other infectious and non- infectious diseases.

  5. Partial blood meal, carbohydrate availability, and blood-feeding postponement effects on human host avidity and DEET repellency in Aedes albopictus (Diptera: Culicidae)

    Science.gov (United States)

    Host avidity and DEET repellency were measured in partially blood fed Aedes albopictus (Skuse) provided 10% sucrose in water, water, or neither when access to a human host was postponed for 1 to 72 h after a partial blood meal. Carbohydrate availability and post-feeding time influenced host avidity...

  6. Prevalence of anti-rubella, anti-measles and anti-mumps IgG antibodies in neonates and pregnant women in Catalonia (Spain) in 2013: susceptibility to measles increased from 2003 to 2013.

    Science.gov (United States)

    Plans, P; de Ory, F; Campins, M; Álvarez, E; Payà, T; Guisasola, E; Compte, C; Vellbé, K; Sánchez, C; Lozano, M J; Aran, I; Bonmatí, A; Carreras, R; Jané, M; Cabero, L

    2015-06-01

    Non-immune neonates and non-immune pregnant women are at risk of developing rubella, measles and mumps infections, including congenital rubella syndrome. We describe the seroepidemiology of measles, mumps and rubella (MMR) in neonates and pregnant women in Catalonia (Spain). Anti-rubella, anti-measles and anti-mumps serum IgG titres were assessed using enzyme-linked immunosorbent assay (ELISA) tests in 353 cord blood samples from neonates of a representative sample of pregnant women obtained in 2013. The prevalence of protective antibody titres in neonates was 96 % for rubella IgG (≥8 IU/ml), 90 % for measles IgG (>300 IU/ml) and 84 % for mumps IgG (>460 EU/ml). Slightly lower prevalences of protective IgG titres, as estimated from the cord blood titres, were found in pregnant women: 95 % for rubella IgG, 89 % for measles IgG and 81 % for mumps IgG. The anti-measles and anti-mumps IgG titres and the prevalences of protective IgG titres against measles and mumps increased significantly (p < 0.001) with maternal age. The prevalence of protective anti-measles IgG titres decreased by 7 % [odds ratio (OR) = 0.15, p < 0.001), the prevalence of protective anti-rubella IgG titres increased by 3 % (OR = 1.80, p < 0.05) and the MMR vaccination coverage (during childhood) in pregnant women increased by 54 % (OR = 2.09, p < 0.001) from 2003 to 2013. We recommend to develop an MMR prevention programme in women of childbearing age based on mass MMR vaccination or MMR screening and vaccination of susceptible women to increase immunity levels against MMR.

  7. Comparison of enzyme-linked immunosorbent assay, radioimmunoassay, complement fixation, anticomplement immunofluorescence and passive haemaglutination techniques for detecting cytomegalovirus IgG antibody

    International Nuclear Information System (INIS)

    Booth, J.C.; Hannington, G.; Bakir, T.M.F.; Stern, H.; Kangro, H.; Griffiths, P.D.; Heath, R.B.

    1982-01-01

    The radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques were found to be comparable in sensitivity and specificity for detecting cytomegalovirus IgG antibody, and 10 to 100 times more sensitive than complement-fixation (CF), anticomplement immunofluorescence (ACIF) and passive haemagglutination (PHA). In screening tests for antibody, the frequency of false-positive and -negative results was 0.6% for RIA and ELISA, 1.5% for CF, 1.6% for ACIF and 3.6% for PHA. PHA was the least satisfactory test, largely because of technical problems. (author)

  8. Immunoglobulin M indirect-fluorescent antibody test for the diagnosis of acute toxoplasmosis during pregnancy in the avidity era: A 14-year experience at the Tuscany Reference Center for Infectious Diseases in Pregnancy, Florence, Italy.

    Science.gov (United States)

    Trotta, Michele; Borchi, Beatrice; Zammarchi, Lorenzo; Sterrantino, Gaetana; Brogi, Michela; Kiros, Seble Tekle; Lorini, Chiara; Bonaccorsi, Guglielmo; Colao, Maria Grazia; Bartoloni, Alessandro

    2016-12-01

    The aim of this study was to evaluate immunoglobulin M indirect-fluorescent antibody test (IgM IFAT) for the diagnosis of acute or chronic Toxoplasma infection in pregnancy. Pregnant women with suspected acute toxoplasmosis referred to the Tuscany Reference Center for Infectious Diseases in Pregnancy during the period 1998-2012 were retrospectively enrolled. All women were tested with a panel of serological tests, including enzyme-linked immunosorbent assay for IgG avidity and IgM IFAT. On the basis of anamnestic, clinical, and serological criteria, pregnant women were classified into three groups: recently infected (RI), latently infected (LI), and doubtful latently infected (DLI). Patients classified as DLI were excluded from the analysis. The association between IgM IFAT (positive or negative) and the diagnosis of infection (acute or chronic) was assessed. Positive predictive value and negative predictive value of the IgM IFAT were calculated. A total of 810 pregnant women were enrolled in the study: 302 in the RI group and 508 in the LI group. Fifty-two women classified as DLI were excluded. IgM IFAT was positive in 172 out of 302 (56.9%) pregnant women in the RI group and in 29 out of 508 (5.7%) in the LI group. The positive predictive value and negative predictive value of IgM IFAT in predicting RI was 85.6% and 78.6%, respectively. IgM IFAT has reasonable sensitivity and specificity in diagnosing recent infection and, mostly in case of borderline avidity test, could be considered as a further aid for an accurate diagnosis of acute toxoplasmosis in pregnancy. © 2016 Japan Society of Obstetrics and Gynecology.

  9. Toxocara Canis IgG Seropositivity in Patients with Chronic Urticaria.

    Science.gov (United States)

    Burak Selek, Mehmet; Baylan, Orhan; Kutlu, Ali; Özyurt, Mustafa

    2015-08-01

    We aimed to investigate IgG antibody levels specific to Toxocara canis (T. canis), a parasite which subsists in dog's intestine, on serum samples obtained from patients with chronic urticaria (CU) to evaluate effective risk in CU etiopathogenesis. In this study, 73 patients diagnosed with CU and 109 healthy individuals as control group, were included. Various factors such as sex, age, education and income, daily hand washing habits, history of dog owning and soil eating were questioned in patient anamnesis. T. canis IgG antibodies were detected using an enzyme linked immunosorbent assay (ELISA) kit prepared with T. canis larval excretory-secretory antigens. Positive results were confirmed with western blot (WB) WB test. We found T. canis IgG positivity in 17.8% (n=13) of patients (n=73) with CU. But we did not observe any T. canis IgG positivity in healthy controls (n=109). Low molecular weight bands (24-35 kDa) were observed in 11 samples in WB analyses while two of the samples were weakly positive. It is revealed that dog owning history increases T. canis seropositivity 12.9 times while insufficient daily hand washing habit (less than six times a day) increases seropositivity 20.7 times. Our study showed that T. canis may trigger CU since we found 17.8% seropositivity in 73 patients with CU and none in 109 healthy individuals. Moreover, various socio-demographic characteristics have been shown to affect T. canis seropositivity in patients with CU.

  10. Structural Basis for Fc[gamma]RIIa Recognition of Human IgG and Formation of Inflammatory Signaling Complexes

    Energy Technology Data Exchange (ETDEWEB)

    Ramsland, Paul A.; Farrugia, William; Bradford, Tessa M.; Sardjono, Caroline Tan; Esparon, Sandra; Trist, Halina M.; Powell, Maree S.; Tan, Peck Szee; Cendron, Angela C.; Wines, Bruce D.; Scott, Andrew M.; Hogarth, P. Mark (Burnet); (Monash); (LICR); (Melbourne)

    2011-09-20

    The interaction of Abs with their specific FcRs is of primary importance in host immune effector systems involved in infection and inflammation, and are the target for immune evasion by pathogens. Fc{gamma}RIIa is a unique and the most widespread activating FcR in humans that through avid binding of immune complexes potently triggers inflammation. Polymorphisms of Fc{gamma}RIIa (high responder/low responder [HR/LR]) are linked to susceptibility to infections, autoimmune diseases, and the efficacy of therapeutic Abs. In this article, we define the three-dimensional structure of the complex between the HR (arginine, R134) allele of Fc{gamma}RIIa (Fc{gamma}RIIa-HR) and the Fc region of a humanized IgG1 Ab, hu3S193. The structure suggests how the HR/LR polymorphism may influence Fc{gamma}RIIa interactions with different IgG subclasses and glycoforms. In addition, mutagenesis defined the basis of the epitopes detected by FcR blocking mAbs specific for Fc{gamma}RIIa (IV.3), Fc{gamma}RIIb (X63-21), and a pan Fc{gamma}RII Ab (8.7). The epitopes detected by these Abs are distinct, but all overlap with residues defined by crystallography to contact IgG. Finally, crystal structures of LR (histidine, H134) allele of Fc{gamma}RIIa and Fc{gamma}RIIa-HR reveal two distinct receptor dimers that may represent quaternary states on the cell surface. A model is presented whereby a dimer of Fc{gamma}RIIa-HR binds Ag-Ab complexes in an arrangement that possibly occurs on the cell membrane as part of a larger signaling assembly.

  11. IFAT and ELISA phase I/phase II as tools for the identification of Q fever chronic milk shedders in cattle.

    Science.gov (United States)

    Lucchese, Laura; Capello, Katia; Barberio, Antonio; Zuliani, Federica; Stegeman, Arjan; Ceglie, Letizia; Guerrini, Eulalia; Marangon, Stefano; Natale, Alda

    2015-08-31

    Q fever is a widespread zoonotic disease caused by Coxiella burnetii. In cattle the bacterial shedding can persist without symptoms for several months and the shedders identification is a critical issue in the control of the infection at herd level. Following the example of the human protocols for the assessment of Q fever infection status, the aim of this study was the evaluation of the antibody response dynamics to phase I and phase II antigens in C. burnetii shedder dairy cows by means of a phase-specific serology, to verify the suitability of the investigated tools in recognising milk shedders. A total of 99 cows were monitored during time and classified on the basis of serological and PCR results in five groups identifying different shedding patterns. The 297 sera collected in three sampling times were tested by means of ELISA IgG for differential phase I and phase II antibodies detection, while a selection of 107 sera were tested by means of phase specific IgM and IgG IFAT. Both ELISA IgG and IFAT IgG highlighted a low reactivity in non-shedder seropositive animals compared to chronic milk shedder animals. ELISA IgG seemed to perform better than IFAT IgG-IgM, showing significant serological differences among groups that allowed recognising specific serological group patterns, in particular for chronic and occasional milk shedders. These results supported the hypothesis that an animal classification based on phase patterns is reasonable, although it needs to be further investigated. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Rapid Enhanced MM3-COPRO ELISA for Detection of Fasciola Coproantigens

    Science.gov (United States)

    Martínez-Sernández, Victoria; Orbegozo-Medina, Ricardo A.; González-Warleta, Marta; Mezo, Mercedes; Ubeira, Florencio M.

    2016-01-01

    ELISA-based methods of detecting Fasciola cathepsins in feces are powerful techniques for diagnosing infections by F. hepatica and F. gigantica. In the last decade, the in-house MM3-COPRO ELISA and its commercial version BIO K 201 (BIO X Diagnostics, Belgium) have been recognized as useful tools for detecting early infections by such trematodes and for monitoring the efficacy of anthelmintic treatments in human and animal species, as they provide some advantages over classic fecal egg counts. However, the sensitivity of MM3-COPRO ELISA can sometimes be compromised by the high variability in the concentration of cathepsins in fecal samples throughout the biological cycle of Fasciola (mainly in cattle) and by differences in the between-batch performance of peroxidase-labeled anti-mouse IgG polyclonal antibodies. To prevent such problems, we investigated whether the incorporation of a commercial streptavidin-polymerized horseradish peroxidase conjugate, in order to reveal bound biotinylated monoclonal antibody MM3, can improve the sensitivity of the MM3-COPRO ELISA. We observed that inclusion of this reagent shifted the previous detection limit of the assay from 0.6 ng/mL to 150 pg/mL and that the modified test is able to identify infection in cows harboring only one fluke. Moreover, we demonstrated that maximal OD values can be achieved with short incubations (30 min each step) at RT with shaking, rather than standard incubations, which significantly accelerates the diagnostic procedure. Finally, we did not find a significant correlation between coproantigen concentration and parasite burden in cattle, which may be due to the low parasite burden (1–10 adult flukes) of the animals used in the present study. As the usefulness of the classic MM3-COPRO test for detecting animal and human infections has already been demonstrated, it is expected that the improvements reported in this study will add new insights into the diagnosis and control of fasciolosis. PMID:27438470

  13. Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I.

    Science.gov (United States)

    van Ree, R; Aalberse, R C

    1995-03-01

    The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a synthetic peptide of its C terminus. When allergen and antibodies were allowed to preincubate, Lol p I induced histamine release was inhibited up to 85% by the antiserum against Lol p I. By omitting preincubation, and thereby more closely mimicking an in vivo situation, up to 55% inhibition was realized. This indicates that allergen-specific IgG can act as 'blocking' antibody without preincubation. Immunization of rabbits with a synthetic C-terminal peptide of Lol p I resulted in antibodies reactive with natural Lol p I. Despite their 100-fold lower avidity for Lol p I (as compared with antinatural Lol p I), these antibodies had the capacity to inhibit Lol p I induced histamine release for > 90% (up to 50% without preincubation). This indicates that it is possible to block histamine release induced by a major allergen with low-avidity IgG antibodies directed against a minor proportion of the allergen (25 amino acids). IgE antibodies from the donors studied were unreactive with this synthetic peptide, indicating that for blocking activity identical epitope specificity of IgE and IgG is not essential. This opens interesting perspectives for application of synthetic peptides in immunotherapy, distinct from their effects on T cell reactivity.

  14. Differential evolution of anti-VAR2CSA- IgG3 in primigravidae and multigravidae pregnant women infected by Plasmodium falciparum

    Science.gov (United States)

    Guitard, Juliette; Cottrell, Gilles; Magnouha, Nellie Moulopo; Salanti, Ali; Li, Tengfei; Sow, Sokhna; Deloron, Philippe; Tuikue Ndam, Nicaise

    2008-01-01

    Background Pregnant women develop protective anti-VSA IgG1 and IgG3 when infected by Plasmodium falciparum. The major target of IgG from serum of infected pregnant women is VAR2CSA. Methods In this study, ELISA was used to compare the level of VAR2CSA DBL5ε- specific IgG subclasses at enrolment and at delivery in a cohort of pregnant women in Senegal. All antibody measures were analysed in relation to placental infection according to parity. Results The results show an interaction between immune response to placental malaria and parity. A higher level of anti- DBL5ε- IgG3 at enrolment and a higher increase between enrolment and delivery were found in primigravidae who presented with uninfected placenta at delivery in comparison to those who presented with an infection of the placenta. However, high antibody level at delivery was associated with the infection of the placenta in multigravidae. Conclusion This high level of IgG3 in uninfected primigravidae suggests a protective role of these antibodies in this susceptible group, highlighting the importance of VAR2CSA in general and of some of its variants still to be defined, in the induction of protective immunity to pregnancy malaria. PMID:18190692

  15. Evaluation of IgY capture ELISA for sensitive detection of alpha hemolysin of Staphylococcus aureus without staphylococcal protein A interference.

    Science.gov (United States)

    Reddy, Prakash Kudumala; Shekar, Aravind; Kingston, Joseph Jeyabalaji; Sripathy, Murali Harishchandra; Batra, Harshvardhan

    2013-05-31

    Staphylococcal protein A (Spa) secreted by all Staphylococcus aureus strains is the major hindrance in development of specific immunoassays for detecting S. aureus antigens, because of its characteristic feature of binding to Fc region of most mammalian immunoglobulins and also to Fab region of certain classes of mammalian immunoglobulins. Immunoglobulin Y (IgY) is the avian equivalent of mammalian IgG which does not have any affinity to Spa. In the present study we report that using chicken egg yolk IgY over mammalian IgG as capture antibody prevents both soluble and surface bound protein A from causing false positives quantified by chicken anti-protein A antibodies. This was demonstrated by development of sandwich ELISA for detection of alpha hemolysin toxin from culture supernatants of S. aureus strains with anti alpha hemolysin IgY as capture and rabbit anti alpha hemolysin IgG as revealing antibody. This indirect sandwich ELISA was evaluated onto a large number of S. aureus isolates recovered from clinical sources for alpha hemolysin secretion. Results of sandwich ELISA were compared with PCR and Western blot analysis. The immunoassay is highly specific and has high sensitivity of detecting less than 1 ng/ml. This procedure is highly effective in eliminating Spa interference and can be extended to detection of other important superantigen toxins of S. aureus. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. ELISA for the Diagnosis of Autoimmune Bullous Disorders

    Directory of Open Access Journals (Sweden)

    Ayşe Akman Karakaş

    2011-06-01

    Full Text Available ELISA has been phrased as Enzyme Linked Immunosorbent Assay. The ELISA provide to detect autoantigens in autoimmune bullous disorders. Therefore, it is assited understanding of the immunopathogenesis of these diseases. Recently, commercial test systems has been developed for the diagnosis and course. The aim of this paper is to review applying investigation of the ELISA for autoimmune bullous disorders.

  17. Advancement Via Individual Determination (AVID) Professional Development as a Predictor of Teacher Leadership in the United States

    Science.gov (United States)

    Watt, Karen M.; Huerta, Jeffery; Mills, Shirley J.

    2010-01-01

    This study expands on the work of Huerta "et al." (2008) by reexamining the relationship between professional development and teacher leadership using more comprehensive measures of each. By confirming Huerta "et al"'s (2008) finding that Advancement Via Individual Determination (AVID) professional development is a significant predictor of teacher…

  18. Alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA applied to dot-ELISA Antígeno de Epimastigota de Trypanosoma cruzi, solubilizado em meio alcalino (ASEA aplicado em dot-ELISA

    Directory of Open Access Journals (Sweden)

    Ana Maria Lissaldo

    1994-04-01

    Full Text Available The alkaline soluble Trypanosoma cruzi epimastigote antigen (ASEA was assessed in dot-ELISA for the diagnosis of Chagas' disease. Serum samples (355 from chagasic and non-chagasic patients were studied, and IgG antibodies to ASEA were found in all patients with chronic Chagas' disease. In non-chagasic patients 95.6% were negative, except for those with leishmaniasis (visceral and mucocutaneous, and some patients from control group reacted in low titers. The data indicate that dot-ELISA using ASEA is suitable for seroepidemiologic surveys to be employed in endemic areas for Chagas' disease.O antígeno de epimastigota do T. cruzi solubilizado em meio alcalino (ASEA foi avaliado em dot-ELISA para o diagnóstico da doença de Chagas. Amostras de soros (355 de chagásicos e não chagásicos foram estudadas e anticorpos IgG contra ASEA foram encontrados em todos os pacientes com a doença de Chagas crônica. Em pacientes não chagásicos, os resultados foram negativos (95,6%, exceto para aqueles com leishmaniose visceral e mucocutânea, e para alguns do grupo controle que reagiram em títulos baixos. Os dados indicam que o ensaio de dot-ELISA, utilizando o ASEA é apropriado para estudos soroepidemiológicos a serem conduzidos em áreas endêmicas da doença de Chagas, tendo em mente a limitação do teste em áreas onde Doença de Chagas e leishmaniose estão presentes.

  19. IgE and allergen-specific immunotherapy-induced IgG4recognize similar epitopes of Bet v 1, the major allergen of birch pollen.

    Science.gov (United States)

    Groh, N; von Loetzen, C S; Subbarayal, B; Möbs, C; Vogel, L; Hoffmann, A; Fötisch, K; Koutsouridou, A; Randow, S; Völker, E; Seutter von Loetzen, A; Rösch, P; Vieths, S; Pfützner, W; Bohle, B; Schiller, D

    2017-05-01

    Allergen-specific immunotherapy (AIT) with birch pollen generates Bet v 1-specific immunoglobulin (Ig)G 4 which blocks IgE-mediated hypersensitivity mechanisms. Whether IgG 4 specific for Bet v 1a competes with IgE for identical epitopes or whether novel epitope specificities of IgG 4 antibodies are developed is under debate. We sought to analyze the epitope specificities of IgE and IgG 4 antibodies from sera of patients who received AIT. 15 sera of patients (13/15 received AIT) with Bet v 1a-specific IgE and IgG 4 were analyzed. The structural arrangements of recombinant (r)Bet v 1a and rBet v 1a _11x , modified in five potential epitopes, were analyzed by circular dichroism and nuclear magnetic resonance spectroscopy. IgE binding to Bet v 1 was assessed by ELISA and mediator release assays. Competitive binding of monoclonal antibodies specific for Bet v 1a and serum IgE/IgG 4 to rBet v 1a and serum antibody binding to a non-allergenic Bet v 1-type model protein presenting an individual epitope for IgE was analyzed in ELISA and western blot. rBet v 1a _11x had a Bet v 1a - similar secondary and tertiary structure. Monomeric dispersion of rBet v 1a _11x was concentration and buffer-dependent. Up to 1500-fold increase in the EC 50 for IgE-mediated mediator release induced by rBet v 1a _11x was determined. The reduction of IgE and IgG 4 binding to rBet v 1a _11x was comparable in 67% (10/15) of sera. Bet v 1a-specific monoclonal antibodies inhibited binding of serum IgE and IgG 4 to 66.1% and 64.9%, respectively. Serum IgE and IgG 4 bound specifically to an individual epitope presented by our model protein in 33% (5/15) of sera. Patients receiving AIT develop Bet v 1a-specific IgG 4 which competes with IgE for partly identical or largely overlapping epitopes. The similarities of epitopes for IgE and IgG 4 might stimulate the development of epitope-specific diagnostics and therapeutics. © 2016 John Wiley & Sons Ltd.

  20. HIV avidity index performance using a modified fourth-generation immunoassay to detect recent HIV infections.

    Science.gov (United States)

    Suligoi, Barbara; Regine, Vincenza; Raimondo, Mariangela; Rodella, Anna; Terlenghi, Luigina; Caruso, Arnaldo; Bagnarelli, Patrizia; Capobianchi, Maria Rosaria; Zanchetta, Nadia; Ghisetti, Valeria; Galli, Claudio

    2017-10-26

    Detecting recent HIV infections is important to evaluate incidence and monitor epidemic trends. We aimed to evaluate the diagnostic performance and accuracy of the avidity index (AI) for discriminating for recent HIV infections. We collected serum samples from HIV-1 positive individuals: A) with known date of infection (midpoint in time between last HIV-negative and first HIV-positive test); B) infected for >1 year. Samples were divided into two aliquots: one diluted with phosphate buffered saline (PBS) and the other with 1 M guanidine. Both aliquots were assayed by the Architect HIV Ag/Ab Combo 4th generation assay (Abbott). We compared AI found in recent (RI=HIV subtype had no impact on AI misclassifications. All individuals in group A reached the AI threshold of 0.80 within 24 months after seroconversion. The AI is an accurate serological marker for discriminating recent from established HIV infections and meets WHO requirements for HIV incidence assays.

  1. Detection of anti-HIV-1 IgG antibodies in whole saliva by GACELISA and Western blot assays.

    Science.gov (United States)

    Matee, M I; Lyamuya, E F; Simon, E; Mbena, E C; Kagoma, C; Samaranayake, L P; Scheutz, F

    1996-05-01

    The present study, based on 158 HIV seropositives and 167 HIV seronegatives, demonstrates that saliva collected with the Omni-SAL device and tested with GACELISA (an IgG antibody capture ELISA) is an effective non-invasive alternative to serum for anti-HIV IgG antibody screening. The study also shows that a conventional serum Western blot kit can be used, with slight modifications, for confirmatory testing of saliva specimens. Collecting saliva with the Omni-SAL device had a very good acceptance rate among Tanzanian subjects, and although this diagnostic method is not yet known by the general public, 65% of the study participants preferred to give saliva instead of blood for HIV testing.

  2. Toxocara canis glycans influence antigen recognition by mouse IgG1 and IgM antibodies.

    Science.gov (United States)

    Długosz, Ewa; Wiśniewski, Marcin

    2016-01-01

    The impact of sugar moieties of Toxocara canis glycoprotein antigens on their recognition by infected mouse antibodies was investigated in this study. Native TES and recombinant Toxocara mucins generated in Pichia pastoris yeast as well as their deglycosylated forms were used in ELISA. TES and recombinant mucins were equally recognized by T. canis infected mouse IgG1 antibodies. IgM immunoglobulins predominantly recognized TES antigens. Among mucins recognition of Tc-MUC-4 was the most significant. Deglycosylation of antigens resulted in significant loss of IgM and IgG1 reactivity to TES, mucins, Tc-MUC-3 and Tc-MUC-4. The presence of sugar moieties had no influence on IgE binding to native or recombinant T. canis antigens. Our results suggest that glycans are involved in epitope formation what should be taken into consideration in production of recombinant helminth antigens for diagnostic purposes.

  3. ELISA technique standardization for strongyloidiasis diagnosis

    International Nuclear Information System (INIS)

    Huapaya, P.; Espinoza, I.; Huiza, A.; Universidad Nacional Mayor de San Marcos, Lima; Sevilla, C.

    2002-01-01

    To standardize ELISA technique for human Strongyloides stercoralis infection diagnosis a crude antigen was prepared using filariform larvae obtained from positive stool samples cultured with charcoal. Harvested larvae were crushed by sonication and washed by centrifugation in order to obtain protein extracts to be used as antigen. Final protein concentration was 600 μg/mL. Several kinds of ELISA plates were tested and antigen concentration, sera dilution, conjugate dilution and cut off were determined to identify infection. Sera from patients with both hyper-infection syndrome and intestinal infection demonstrated by parasitological examination were positive controls and sera from people living in non-endemic areas with no infection demonstrated by parasitological examination were negative controls. Best values were 5 μg/mL for antigen, 1/64 for sera, 1/1000 for conjugate; optical density values for positive samples were 1,2746 (1,1065 - 1,4206, DS = 0,3284) and for negative samples 0,4457 (0,3324 - 0,5538, DS = 0,2230). Twenty sera samples from positive subjects and one hundred from negative subjects were examined, obtaining 90% sensitivity and 88% specificity. The results show this technique could be useful as strongyloidiasis screening test in population studies

  4. Application of 60Co γ-ray irradiated polystyrene microplate for anti-HCV ELISA

    International Nuclear Information System (INIS)

    Feng Bo; Zeng Hongyan; Tang Yufang; Wang Lu

    2005-01-01

    In order to explore the effect of 60 Co γ-ray irradiation on minor polypeptides absorption of polystyrene microplate, an indirect ELISA detection of anti-HCV was established, 60 Co γ-ray irradiated polystyrene microplates and the controls (without irradiation or UV-irradiated) were applied to absorb recombinant HCV antigens respectively. Cooperated with Bovine antihuman IgG labelled HRP, their related indices of sensitivity, specificity, homogeneity and stability were determinate. The results indicated that, optimum dose of the γ-ray irradiation is 8 kGy, and compared with the controls, detection sensitivity and homogeneity of the polystyrene microplate irradiated to 8 kGy could be improved markedly. (authors)

  5. The influence of dengue virus serotype-2 infection on Aedes aegypti (Diptera: Culicidae motivation and avidity to blood feed.

    Directory of Open Access Journals (Sweden)

    Rafael Maciel-de-Freitas

    Full Text Available BACKGROUND: Dengue virus (DENV is transmitted by Aedes aegypti, a species that lives in close association with human dwellings. The behavior of DENV-infected mosquitoes needs further investigation, especially regarding the potential influence of DENV on mosquito biting motivation and avidity. METHODOLOGY/PRINCIPAL FINDINGS: We orally challenged 4-5 day-old Ae. aegypti females with a low passage DENV serotype -2 (DENV-2 to test whether the virus influences motivation to feed (the likelihood that a mosquito obtains a blood-meal and the size of its blood meal and avidity (the likelihood to re-feed after an interrupted first blood-meal. To assay motivation, we offered mosquitoes an anesthetized mouse for 2, 3, 4 or 5 minutes 7 or 14 days after the initial blood meals and measured the time they started feeding. 60.5% of the unexposed mosquitoes fed on the mouse, but only 40.5% of the positive ones did. Exposed but negative mosquitoes behaved similarly to unexposed ones (55.0% feeding. Thus DENV-2 infection decreased the mosquitoes' motivation to feed. To assay avidity, we offered the same mosquitoes a mouse two hours after the first round of feeding, and we measured the time at which they started probing. The exposed (positive or negative mosquitoes were more likely to re-feed than the unexposed ones and, in particular, the size of the previous blood-meal that kept mosquitoes from re-feeding was larger in the exposed than in the unexposed mosquitoes. Thus, DENV-2 infection increased mosquito avidity. CONCLUSIONS/SIGNIFICANCE: DENV-2 significantly decreased the mosquitoes' motivation to feed, but increased their avidity (even after taking account the amount of blood previously imbibed. As these are important components of transmission, we expect that the changes of the blood-feeding behaviour impact the vectorial capacity Ae. aegypti for dengue.

  6. The influence of dengue virus serotype-2 infection on Aedes aegypti (Diptera: Culicidae) motivation and avidity to blood feed.

    Science.gov (United States)

    Maciel-de-Freitas, Rafael; Sylvestre, Gabriel; Gandini, Mariana; Koella, Jacob C

    2013-01-01

    Dengue virus (DENV) is transmitted by Aedes aegypti, a species that lives in close association with human dwellings. The behavior of DENV-infected mosquitoes needs further investigation, especially regarding the potential influence of DENV on mosquito biting motivation and avidity. We orally challenged 4-5 day-old Ae. aegypti females with a low passage DENV serotype -2 (DENV-2) to test whether the virus influences motivation to feed (the likelihood that a mosquito obtains a blood-meal and the size of its blood meal) and avidity (the likelihood to re-feed after an interrupted first blood-meal). To assay motivation, we offered mosquitoes an anesthetized mouse for 2, 3, 4 or 5 minutes 7 or 14 days after the initial blood meals and measured the time they started feeding. 60.5% of the unexposed mosquitoes fed on the mouse, but only 40.5% of the positive ones did. Exposed but negative mosquitoes behaved similarly to unexposed ones (55.0% feeding). Thus DENV-2 infection decreased the mosquitoes' motivation to feed. To assay avidity, we offered the same mosquitoes a mouse two hours after the first round of feeding, and we measured the time at which they started probing. The exposed (positive or negative) mosquitoes were more likely to re-feed than the unexposed ones and, in particular, the size of the previous blood-meal that kept mosquitoes from re-feeding was larger in the exposed than in the unexposed mosquitoes. Thus, DENV-2 infection increased mosquito avidity. DENV-2 significantly decreased the mosquitoes' motivation to feed, but increased their avidity (even after taking account the amount of blood previously imbibed). As these are important components of transmission, we expect that the changes of the blood-feeding behaviour impact the vectorial capacity Ae. aegypti for dengue.

  7. Image-based ELISA on an activated polypropylene microtest plate--a spectrophotometer-free low cost assay technique.

    Science.gov (United States)

    Parween, Shahila; Nahar, Pradip

    2013-10-15

    In this communication, we report ELISA technique on an activated polypropylene microtest plate (APPµTP) as an illustrative example of a low cost diagnostic assay. Activated test zone in APPµTP binds a capture biomolecule through covalent linkage thereby, eliminating non-specific binding often prevalent in absorption based techniques. Efficacy of APPµTP is demonstrated by detecting human immunoglobulin G (IgG), human immunoglobulin E (IgE) and Aspergillus fumigatus antibody in patient's sera. Detection is done by taking the image of the assay solution by a desktop scanner and analyzing the color of the image. Human IgE quantification by color saturation in the image-based assay shows excellent correlation with absorbance-based assay (Pearson correlation coefficient, r=0.992). Significance of the relationship is seen from its p value which is 4.087e-11. Performance of APPµTP is also checked with respect to microtiter plate and paper-based ELISA. APPµTP can quantify an analyte as precisely as in microtiter plate with insignificant non-specific binding, a necessary prerequisite for ELISA assay. In contrast, paper-ELISA shows high non-specific binding in control sera (false positive). Finally, we have carried out ELISA steps on APPµTP by ultrasound waves on a sonicator bath and the results show that even in 8 min, it can convincingly differentiate a test sample from a control sample. In short, spectrophotometer-free image-based miniaturized ELISA on APPµTP is precise, reliable, rapid, and sensitive and could be a good substitute for conventional immunoassay procedures widely used in clinical and research laboratories. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Diagnostic Value of ELISA Tests for the Detection of Specific Antibodies in Cats and Rabbits with Dermatophytosis

    Directory of Open Access Journals (Sweden)

    Marinka Drobnič-Košorok

    2002-01-01

    Full Text Available Two indirect ELISA tests developed for the detection of specific IgG in cats and rabbits, infected with M. canis and T. mentagrophytes, respectively, were evaluated and compared. The levels of specific antibodies were determined in sera of 20 cats and 25 rabbits naturally infected with M. canis and T. mentagrophytes, respectively. Infection was confirmed by the results of fungal culture. Blood samples from 12 cats and 17 rabbits, previously unexposed to dermatophytes, served as negative controls. A significant increase in the level of specific antibodies in groups of infected animals was demonstrated. Sensitivity, specificity and predictive values of a positive and a negative test were determined to evaluate the diagnostic potential. ELISA for the detection of specific antibodies in cats infected with M. canis (ELISA-cats test exhibited 75.0 % of sensitivity at 91.7 % of specificity, whereas the test for the detection of specific antibodies in rabbits, infected with T. mentagrophytes (ELISA-rabbits test is highly sensitive (96.0 % and highly specific (94.1 %, confirming its encouraging diagnostic potential. The cross-reactivity of fungal antigens was tested by performing the assays with antigens M. canis, T. mentagrophytes, M. pachydermatis and A. fumigatus. There were no significant indications of cross-reactions in the test T. mentagrophytes-rabbits, whereas strong cross-reaction between dermatophyte antigens was observed in the test M. canis-cats.

  9. IgG antibody response against salivary gland antigens from Oestrus ovis L. larvae (Diptera: Oestridae) in experimentally and naturally infected goats.

    Science.gov (United States)

    Angulo-Valadez, C E; Cepeda-Palacios, R; Ascencio, F; Jacquiet, Ph; Dorchies, Ph; Ramírez-Orduña, J M; López, M A

    2009-05-12

    The aims of this study were to analyze the systemic IgG responses against third-instar salivary gland (L3SG) antigens by ELISA in Oestrus ovis experimentally infected kids (EIK) and in naturally exposed adult goats (NEG). Firstly, kids (n=4 per group) were assigned to receive intranasally 0, 12, 24, 36, and 48 first-instars in experimental infections. Blood samples were taken from EIK at Days 0, 14, 42 and 67 post-infection. At necropsy (Day 67), larval number and developmental instars were recorded. In an epidemiological study, blood serum samples were collected from 448 grazing NEG (n=20 flocks) in Baja California Sur, Mexico. Results showed that larval establishment rate was similar in EIK groups. Systemic IgG response reached the threshold after Day 42, but humoral response was not statistically different among EIK groups receiving experimental infections. In NEG, all surveyed flocks (100%) showed specific systemic IgG antibodies to L3SG antigens and the overall goat oestrosis prevalence was 59.2%. In conclusion, larval L3SG antigens were effective in detection of specific systemic IgG antibodies against O. ovis infected kids and goats by ELISA.

  10. Purification, characterization and ELISA detection of mink immunoglobulins

    DEFF Research Database (Denmark)

    Martel, Cyril Jean-Marie; Aasted, Bent

    2008-01-01

    This study describes easy purification methods for mink IgG, IgA and IgM immunoglobulins. IgG and IgM were purified from normal mink serum, while IgA was purified from mink bile from healthy animals. By SDS-polyacrylamid-gel-electrophoresis  (SDS-PAGE) and immunoblotting under reducing conditions...

  11. Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool

    Directory of Open Access Journals (Sweden)

    Zoheiry Mona K

    2011-09-01

    Full Text Available Abstract Background This research was carried out to develop a reliable monoclonal antibody (MoAb-based sandwich enzyme linked immunosorbent assay (ELISA for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. Methods From a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags, a pair (12B/11D/3F and 10A/9D/10G was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA. Results The two MoAbs were of the IgG1 and IgG2a subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p Conclusions These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.

  12. Development of ELISA for the detection of transgenic vegetative insecticidal protein in GM crops/produce.

    Science.gov (United States)

    Kumar, R

    2012-01-11

    In the process of the development of insect-resistant genetically modified (GM) crops and also to evaluate the consistency in the expression of toxin under field conditions, immunological assays are commonly being used. An immunoassay was developed to support the labelling of vegetative insecticidal protein (Vip3A)-based GM produce. The developed ELISA for the measurement of Vip3A is a triple antibody sandwich procedure utilising a polyclonal capture antibody (mouse anti-Vip3A) and a polyclonal detection antibody (rabbit anti-Vip3A) followed by use of a third HRP-conjugated anti-species antibody (goat anti-rabbit IgG). The limit of detection limit of the ELISA assay was 16 ng ml(-1) with a linear quantification range from approximately 31 to 500 ng ml(-1) of Vip3A protein. Furthermore, the assay was in-house validated with GM brinjal samples. The assay was specific, sensitive and reproducible, which can be helpful to detect and track down the spread of unapproved and intentionally/unintentionally released GM produce harbouring Vip protein.

  13. A sensitive sandwich ELISA for the detection of trace amounts of cashew (Anacardium occidentale L.) nut in foods.

    Science.gov (United States)

    Wei, Yanhong; Sathe, Shridhar K; Teuber, Suzanne S; Roux, Kenneth H

    2003-05-21

    Trace amounts of cashew nut protein can provoke severe allergic reactions in sensitive patients. Consequently, commercial food processors and regulatory agencies must be vigilant to prevent cashew nut cross-contamination among foods and ensure proper labeling. Toward this end, we have developed a sandwich enzyme-linked immunosorbent (ELISA) to detect the predominant cashew protein fraction (anacardein or cashew major protein, CMP) that can be extracted in aqueous buffer from food matrixes. Protein G-purified goat antiwhole cashew extract IgG and rabbit anti-CMP IgG were used as capture and secondary antibodies, respectively. Immunoadsorption against several nut and seed proteins significantly minimized the inherent cross-reactivity of these reagents. Food samples spiked with cashew flour and CMP were extracted and tested in a sandwich ELISA where standard curves were based on reactivity with CMP. The assay was optimized to detect as little as 20 ng/mL (0.02 ppm) of CMP and was successfully used to quantify CMP, and thus cashew, in various food matrixes.

  14. Specificity and Effector Functions of Human RSV-Specific IgG from Bovine Milk.

    Directory of Open Access Journals (Sweden)

    Gerco den Hartog

    Full Text Available Respiratory syncytial virus (RSV infection is the second most important cause of death in the first year of life, and early RSV infections are associated with the development of asthma. Breastfeeding and serum IgG have been shown to protect against RSV infection. Yet, many infants depend on bovine milk-based nutrition, which at present lacks intact immunoglobulins.To investigate whether IgG purified from bovine milk (bIgG can modulate immune responses against human RSV.ELISAs were performed to analyse binding of bIgG to human respiratory pathogens. bIgG or hRSV was coated to plates to assess dose-dependent binding of bIgG to human Fcγ receptors (FcγR or bIgG-mediated binding of myeloid cells to hRSV respectively. S. Epidermidis and RSV were used to test bIgG-mediated binding and internalisation of pathogens by myeloid cells. Finally, the ability of bIgG to neutralise infection of HEp2 cells by hRSV was evaluated.bIgG recognised human RSV, influenza haemagglutinin and Haemophilus influenza. bIgG bound to FcγRII on neutrophils, monocytes and macrophages, but not to FcγRI and FcγRIII, and could bind simultaneously to hRSV and human FcγRII on neutrophils. In addition, human neutrophils and dendritic cells internalised pathogens that were opsonised with bIgG. Finally, bIgG could prevent infection of HEp2 cells by hRSV.The data presented here show that bIgG binds to hRSV and other human respiratory pathogens and induces effector functions through binding to human FcγRII on phagocytes. Thus bovine IgG may contribute to immune protection against RSV.

  15. Comparison of IgM Capture Enzyme- Linked Immunosorbent Assay (ELISA) using Inhouse method and commercially available MRL kit for serological confirmation of dengue infection

    International Nuclear Information System (INIS)

    Hapugoda, D.M.; De Silva R, Nilanthi; Abeywickreme, W.; Gunasena, Sunethra; Prithimala, L.D.; Jayawardene, S.L.G.J.; Kumari, Thamara

    2003-01-01

    Laboratory diagnosis of dengue infection is important for the management of the patients. In this study igM capture ELISA using an inhouse method and commercially available kit (MRL diagnostics,USA) was compared to detect diagnostic capability of Inhouse IgM ELISA for provision of diagnostic facilities to the public at an affordable cost. Eighty acute and convalescent serum samples were collected from serologically confirmed dengue patients. Serological confirmation of patients were performed by Haemagglutination Inhibition (HI) assay, gold standard assay for dengue on paired serum samples. All collected acute and convalescent sera were tested by IgM ELISA using the inhouse method and MRL kit. Antigen and conjugate for the inhouse IgM method were prepared in the laboratory. A cocktail of four dengue antigens containing 25 Antigen ELISA units of each type was prepared and used as the assay antigen. Conjugate was prepared using a serum sample with high dengue Anti flavi IgG antibody titre conjugated with Horseradish peroxidase. A prospective study of both IgM ELISA assays were performed using 113 acute sera collected from dengue suspected cases. Overall results showed that 46% and 52% acute sera collected from dengue confirmed patients were positive by inhouse ELISA assay and MRL kits respectively. In the prospective study done using acute sera collected from dengue suspected patients showed that 44% and 52% were positive by inhouse ELISA assay and MRL kits. There was no significant difference in positivity between these two assays. (P=0.18). Inhouse IgM ELISA can be used for provision of laboratory diagnosis of dengue virus infection more than 5 days. The assay is 10 times less costly than using MRL kits as assay antigen and conjugate can be prepared easily in the laboratory

  16. Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus.

    Science.gov (United States)

    Mealey, Robert H; Zhang, Baoshan; Leib, Steven R; Littke, Matt H; McGuire, Travis C

    2003-09-01

    Equine infectious anemia virus (EIAV) is a lentivirus that causes persistent infections in horses. We hypothesized that high-avidity CTL specific for nonvariable epitopes might be associated with low viral load and minimal disease in EIAV-infected horses. To test this hypothesis, memory CTL (CTLm) responses were analyzed in two infected horses with high plasma viral loads and recurrent disease (progressors), and in two infected horses with low-to-undetectable viral loads and mild disease (nonprogressors). High-avidity CTLm in one progressor recognized an envelope gp90 epitope, and the data documented for the first time in EIAV that viral variation led to CTL escape. Each of the nonprogressors had high-to-moderate avidity CTLm directed against epitopes within Rev, including the nuclear export and nuclear localization domains. These results suggested that the epitope specificity of high- and moderate-avidity CTLm was an important determinant for disease outcome in the EIAV-infected horses examined.

  17. Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus

    International Nuclear Information System (INIS)

    Mealey, Robert H.; Zhang Baoshan; Leib, Steven R.; Littke, Matt H.; McGuire, Travis C.

    2003-01-01

    Equine infectious anemia virus (EIAV) is a lentivirus that causes persistent infections in horses. We hypothesized that high-avidity CTL specific for nonvariable epitopes might be associated with low viral load and minimal disease in EIAV-infected horses. To test this hypothesis, memory CTL (CTLm) responses were analyzed in two infected horses with high plasma viral loads and recurrent disease (progressors), and in two infected horses with low-to-undetectable viral loads and mild disease (nonprogressors). High-avidity CTLm in one progressor recognized an envelope gp90 epitope, and the data documented for the first time in EIAV that viral variation led to CTL escape. Each of the nonprogressors had high-to-moderate avidity CTLm directed against epitopes within Rev, including the nuclear export and nuclear localization domains. These results suggested that the epitope specificity of high- and moderate-avidity CTLm was an important determinant for disease outcome in the EIAV-infected horses examined

  18. Epitope specificity is critical for high and moderate avidity cytotoxic T lymphocytes associated with control of viral load and clinical disease in horses with equine infectious anemia virus

    Science.gov (United States)

    Mealey, Robert H.; Zhang, Baoshan; Leib, Steven R.; Littke, Matt H.; McGuire, Travis C.

    2012-01-01

    Equine infectious anemia virus (EIAV) is a lentivirus that causes persistent infections in horses. We hypothesized that high-avidity CTL specific for nonvariable epitopes might be associated with low viral load and minimal disease in EIAV-infected horses. To test this hypothesis, memory CTL (CTLm) responses were analyzed in two infected horses with high plasma viral loads and recurrent disease (progressors), and in two infected horses with low-to-undetectable viral loads and mild disease (nonprogressors). High-avidity CTLm in one progressor recognized an envelope gp90 epitope, and the data documented for the first time in EIAV that viral variation led to CTL escape. Each of the nonprogressors had high-to-moderate avidity CTLm directed against epitopes within Rev, including the nuclear export and nuclear localization domains. These results suggested that the epitope specificity of high- and moderate-avidity CTLm was an important determinant for disease outcome in the EIAV-infected horses examined. PMID:12954220

  19. HLA-A(∗)02, gender and tobacco smoking, but not multiple sclerosis, affects the IgG antibody response against human herpesvirus 6.

    Science.gov (United States)

    Engdahl, Elin; Gustafsson, Rasmus; Ramanujam, Ryan; Sundqvist, Emilie; Olsson, Tomas; Hillert, Jan; Alfredsson, Lars; Kockum, Ingrid; Fogdell-Hahn, Anna

    2014-06-01

    Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system. Both genetic and environmental factors contribute to disease susceptibility and two viruses associated with MS are human herpesvirus (HHV)-6A and HHV-6B, together referred to as HHV-6. This study characterized the plasma IgG antibody response against HHV-6 in MS patients (n=446) and healthy controls (n=487), and the relationship between MS susceptibility factors and the anti-HHV-6 response was investigated. In addition, 134 samples were further investigated for IgG against the early HHV-6 antigen p41. Antibody levels were measured with ELISA. The overall seroprevalence against HHV-6 was 90%, with no significant difference in positivity or levels between MS patients and controls. Interestingly, carriership of HLA-A(∗)02 and tobacco smoking was associated with lower anti-HHV-6 IgG levels (p=0.0017 and p=0.026 respectively), whereas females sex was associated with higher levels (p=0.0090). No difference in IgG titers against p41 was observed between MS patients and controls. In conclusion, the IgG response against HHV-6 was associated with several factors that have previously been associated with MS susceptibility, possibly reflecting a relation between autoimmunity and how the immune system handles viral infections. Copyright © 2014 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

  20. Modified dot-ELISA for diagnosis of human trichinellosis.

    Science.gov (United States)

    Taher, Eman E; Méabed, Eman M H; El Akkad, Dina M H; Kamel, Nancy O; Sabry, Maha A

    2017-06-01

    This study aimed to modify Dot-Enzyme-linked immunosorbent assay (dot-ELISA) for the diagnosis of human trichinellosis and to compare its performance with indirect ELISA and Western-blot assay (EITB). A total of 175 human serum samples were enrolled in the study. Indirect ELISA was used for the primary diagnosis. EITB versus fractionated 1st larval stage excretory-secretory antigens (TL-1 ESA) revealed three specific protein fractions at MW of 45, 50, and 55 kDa (kDa). Dot-ELISA was performed in two ways. In the first one, sera were dotted on the separated three specific protein fractions, while in the second one the three fractions were eluted, concentrated at one pooled antigen that used in classic dot-ELISA. Both types of dot-ELISA proved absolute (100%) sensitivity and specificity in comparison with the gold standard EITB reaction. While sensitivity of ELISA was 100% and its specificity was 79.5%. The fraction at 45 kDa was the most sensitive one. The use of the pooled antigen improved the test results. The described dot-ELISA is an easy applicable diagnostic tool gathering the benefits of both ELISA and EITB. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Immune response against Treponema spp. and ELISA detection of digital dermatitis.

    Science.gov (United States)

    Gomez, A; Anklam, K S; Cook, N B; Rieman, J; Dunbar, K A; Cooley, K E; Socha, M T; Döpfer, D

    2014-01-01

    The objective of this longitudinal study was to evaluate the immune response against Treponema spp. infection in dairy heifers affected with digital dermatitis (DD). In addition, the accuracy of an indirect ELISA detecting anti-Treponema IgG antibodies in identifying clinical DD status has been assessed. A cohort of 688 pregnant Holstein heifers was evaluated at least 3 times before calving during a period of 6 mo. Complete clinical assessment of DD presence on the back feet of each heifer and blood extraction were performed in a stand-up chute. Digital dermatitis cases were characterized by the M-stage classification system and size and level of skin proliferation. An ELISA was performed on blood serum samples obtained from a subcohort of 130 heifers. For description purposes, the animals were classified by the number of clinical cases experienced during the study period as type I (no clinical cases were observed), type II (only 1 acute clinical case diagnosed), and type III (at least 2 acute clinical cases diagnosed). Multivariable repeated-measures models were used to evaluate the immune response against Treponema spp. infection. A binormal Bayesian model for the ELISA data without cut-point values was used to assess the accuracy of the ELISA as a diagnostic tool. Animals that never experienced a DD event throughout the study kept a constant low level of antibody titer. A 56% increase in mean ELISA titer was observed in heifers upon a first clinical DD case diagnosis. After topical treatment of an acute DD case with oxytetracycline, the antibody titer decreased progressively in type II heifers, achieving mean levels of those observed in healthy cows after a mean of 223 d. Surprisingly, antibody titer was not increased in the presence of M1 (DD lesion <20mm in diameter surrounded by healthy skin) and M4.1 (DD lesion <20mm in diameter embedded in a circumscribed dyskeratotic or proliferative skin alteration) DD stages. Type III cows showed a slight increase in

  2. Enhanced change detection performance reveals improved strategy use in avid action video game players.

    Science.gov (United States)

    Clark, Kait; Fleck, Mathias S; Mitroff, Stephen R

    2011-01-01

    Recent research has shown that avid action video game players (VGPs) outperform non-video game players (NVGPs) on a variety of attentional and perceptual tasks. However, it remains unknown exactly why and how such differences arise; while some prior research has demonstrated that VGPs' improvements stem from enhanced basic perceptual processes, other work indicates that they can stem from enhanced attentional control. The current experiment used a change-detection task to explore whether top-down strategies can contribute to VGPs' improved abilities. Participants viewed alternating presentations of an image and a modified version of the image and were tasked with detecting and localizing the changed element. Consistent with prior claims of enhanced perceptual abilities, VGPs were able to detect the changes while requiring less exposure to the change than NVGPs. Further analyses revealed this improved change detection performance may result from altered strategy use; VGPs employed broader search patterns when scanning scenes for potential changes. These results complement prior demonstrations of VGPs' enhanced bottom-up perceptual benefits by providing new evidence of VGPs' potentially enhanced top-down strategic benefits. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Interference of daratumumab with pretransfusion testing, mimicking a high-titer, low avidity like antibody

    Directory of Open Access Journals (Sweden)

    Mei-Hwa Lin

    2017-01-01

    Full Text Available Daratumumab is a monoclonal immunoglobulin against CD38 and has been approved for treating patients with refractory multiple myeloma. The presence of daratumumab in the sera can interfere with pretransfusion testing due to the weakly expression of CD38 on red cells. The reactivity could be mistaken as autoantibody (if autocontrol is positive or alloantibody (if autocontrol is negative. We present a case that demonstrates daratumumab could mimic a high titer low avidity (HTLA alloantibody. A 34-year-old male patient of refractory myeloma was recruited in phase three clinical trial involving daratumumab. Samples were sent to the blood bank for pretransfusion testing. Without knowledge of patient having used daratumumab, we mistook the reactivity in the patient's sera as an HTLA antibody due to the results of negative autocontrol and high titers of antibody activity. Antibody screen showed a panreactive pattern and the reactivity against screening cells was up to a titer of 1: 1240. The reactivity was weaker against cord cells than adult cells, became weaker against ZZAP-treated cells and became negative against DDT-treated cells. A discussion with attending physician finally revealed the reactivity was due to the interference caused by daratumumab. The case demonstrates good communication is essential in performing pretransfusion testing for patients receiving daratumumab and other new biological regimens that can interfere with compatibility test.

  4. Interference of daratumumab with pretransfusion testing, mimicking a high-titer, low avidity like antibody.

    Science.gov (United States)

    Lin, Mei-Hwa; Liu, Fei-Yun; Wang, Hsiu-Mien; Cho, Hsin-Ching; Lo, Shyh-Chyi

    2017-01-01

    Daratumumab is a monoclonal immunoglobulin against CD38 and has been approved for treating patients with refractory multiple myeloma. The presence of daratumumab in the sera can interfere with pretransfusion testing due to the weakly expression of CD38 on red cells. The reactivity could be mistaken as autoantibody (if autocontrol is positive) or alloantibody (if autocontrol is negative). We present a case that demonstrates daratumumab could mimic a high titer low avidity (HTLA) alloantibody. A 34-year-old male patient of refractory myeloma was recruited in phase three clinical trial involving daratumumab. Samples were sent to the blood bank for pretransfusion testing. Without knowledge of patient having used daratumumab, we mistook the reactivity in the patient's sera as an HTLA antibody due to the results of negative autocontrol and high titers of antibody activity. Antibody screen showed a panreactive pattern and the reactivity against screening cells was up to a titer of 1: 1240. The reactivity was weaker against cord cells than adult cells, became weaker against ZZAP-treated cells and became negative against DDT-treated cells. A discussion with attending physician finally revealed the reactivity was due to the interference caused by daratumumab. The case demonstrates good communication is essential in performing pretransfusion testing for patients receiving daratumumab and other new biological regimens that can interfere with compatibility test.

  5. Current Concept of IgG4-Related Disease.

    Science.gov (United States)

    Okazaki, Kazuichi; Umehara, Hisanori

    2017-01-01

    IgG4-related disease (IgG4-RD) is a fibroinflammatory disease of unknown etiology, which is characterized by a tendency to form tumefactive lesions, increased serum levels of IgG4, and massive infiltration of IgG4-positive plasma cells with storiform fibrosis and/or obliterative phlebitis. Patients with IgG4-RD have frequently multiorgan involvements such as the pancreas, biliary tree, salivary glands, periorbital tissues, kidneys, lungs, lymph nodes, and retroperitoneum. IgG4-RD mainly affects middle-aged to elderly men except for involvement in lachrymal and salivary glands, so-called Mikulicz's disease. The clinical manifestations of IgG4-RD depend on individually involved organs and respond well to steroid, but the prognosis still remains unclear. Some patients develop serious complications such as obstructive jaundice due to hepatic, gallbladder, or pancreatic lesions; hydronephrosis due to retroperitoneal fibrosis; or respiratory symptoms due to pulmonary lesions. Nomenclatures of individual organ manifestation of IgG4-RD have been internationally consented.

  6. Flexibility and conformational change of IgG molecule

    International Nuclear Information System (INIS)

    Alpert, Y.; Ostanevich, Yu.M.

    1982-12-01

    The dynamic behaviour of pig anti-Dnp-immunoglobulin (IgG) investigated by the neutron spin echo technique gave evidence of internal motion of a biological macromolecule. It is suggested that this motion belongs to the wobbling of the Fab parts of the investigated IgG molecule around its so called hinge region. (author)

  7. Seropositivity and determinants of immunoglobulin-G (IgG ...

    African Journals Online (AJOL)

    Background: This study is the first documented prevalence of IgG antibody against HSV-1&-2 in Port Harcourt, Nigeria and thus provides baseline data for future in-depth studies on HSV infection in South-South, Nigeria. Objective: This study determined the seropositivity and determinants of serum IgG antibody against ...

  8. THE RELATIONSHIP OF GAMMA IMMUNOGLOBIN (IgG)

    African Journals Online (AJOL)

    2013-07-31

    Jul 31, 2013 ... transferred IgG or IgG density (IgGρ) was estimated to find out if it has any relevance to the condition of an infant. 1-5 minutes after birth or APGAR score which ... Microvasculature derived cells (with the aid of a dissecting microscope) were used in preference to endothelial cells isolated from large vessels ...

  9. IgG4-Related Disease: A Multispecialty Condition

    Directory of Open Access Journals (Sweden)

    Iuri Usêda Santana

    2014-01-01

    Full Text Available IgG4-related disease (IgG4-RD is a recently recognized group of conditions, characterized by tumor-like swelling of involved organs, lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells, variable degrees of fibrosis, and elevated serum IgG4 concentrations. Currently IgG4-RD is recognized as a systemic condition that can affect several organs and tissues. Herein we report the case of a 34-year-old male patient who was admitted to our hospital with diffuse abdominal pain, weight loss, and painful stiffness in his neck. He had a history of tumoral mass of the left maxillary region, right palpebral ptosis with protrusion of the eyeball, and chronic dry cough for about 6 years. Laboratory tests revealed polyclonal hypergammaglobulinemia and increased serum IgG4 levels. Immunohistochemical staining of the maxillary biopsy was compatible with IgG4-RD. He had an excellent response to corticosteroid therapy. This case highlights that IgG4-RD should be included in the differential diagnosis with multisystem diseases.

  10. Development & validation of a quantitative anti-protective antigen IgG enzyme linked immunosorbent assay for serodiagnosis of cutaneous anthrax

    Directory of Open Access Journals (Sweden)

    N Ghosh

    2015-01-01

    Full Text Available Background & objectives: Anthrax caused by Bacillus anthracis is primarily a disease of herbivorous animals, although several mammals are vulnerable to it. ELISA is the most widely accepted serodiagnostic assay for large scale surveillance of cutaneous anthrax. The aims of this study were to develop and evaluate a quantitative ELISA for determination of IgG antibodies against B. anthracis protective antigen (PA in human cutaneous anthrax cases. Methods: Quantitative ELISA was developed using the recombinant PA for coating and standard reference serum AVR801 for quantification. A total of 116 human test and control serum samples were used in the study. The assay was evaluated for its precision, accuracy and linearity. Results: The minimum detection limit and lower limit of quantification of the assay for anti-PA IgG were 3.2 and 4 µg/ml, respectively. The serum samples collected from the anthrax infected patients were found to have anti-PA IgG concentrations of 5.2 to 166.3 µg/ml. The intra-assay precision per cent CV within an assay and within an operator ranged from 0.99 to 7.4 per cent and 1.7 to 3.9 per cent, respectively. The accuracy of the assay was high with a per cent error of 6.5 - 24.1 per cent. The described assay was found to be linear between the range of 4 to 80 ng/ml (R [2] =0.9982; slope=0.9186; intercept = 0.1108. Interpretation & conclusions: The results suggested that the developed assay could be a useful tool for quantification of anti-PA IgG response in human after anthrax infection or vaccination.

  11. Evaluation by ELISA of Anisakis simplex Larval Antigen Purified by Affinity Chromatography

    Directory of Open Access Journals (Sweden)

    Rodero M

    2002-01-01

    Full Text Available In order to improve the specificity and sensitivity of the techniques for the human anisakidosis diagnosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis simplex third-stage larvae (L3 has been developed. New Zealand rabbits were immunized with A. simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG specific antibodies were isolated by means of protein A-Sepharose CL-4B beads columns. IgG anti-A. simplex and -A. suum were coupled to CNBr-activated Sepharose 4B. For the purification of the larval A. simplex antigens, these were loaded into the anti-A. simplex column and bound antigens eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. simplex specific proteins were loaded into the anti-A. suum column. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis were carried out. Further, we studied the different responses by ELISA to the different antigenic preparations of A. simplex used, observing their capability of discriminating among the different antisera raised in rabbits (anti-A. simplex, anti-A. suum, anti-T. canis. The discriminatory capability with the anti-T. canis antisera was good using the larval A. simplex crude extract (CE antigen. When larval A. simplex CE antigen was loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with A. simplex CE antigen, its capability for discriminate between A. simplex and A. suum was improved, increasing in the case of T. canis. The best results were obtained using larval A. simplex CE antigen loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with adult A. suum CE antigen. When we compared the different serum dilution and antigenic concentration, we selected the working serum dilution of 1/400 and 1 mg/ml of antigenic concentration.

  12. Seroprevalence of IgG anti-Toxocara canis antibodies and anti-Fasciola sp. antibodies in patients with urticaria.

    Science.gov (United States)

    Dal, T; Ciçek, M; Uçmak, D; Akkurt, M; Tekin, A; Dal, M S; Tekin, R; Kalkanl, S T

    2013-01-01

    Chronic urticaria (CU) is defined by recurrent episodes occurring at least twice a week for 6 weeks and generally characterized by the rapid appearance of wheals and/or angioedema and may be associated with parasitic infections. We aimed to investigate the seroprevalance of Toxocara canis and Fasciola species in patients with CU. We included 55 patients (in age 16-55) with urticaria admitted in study. As a control group we recruited 30 healthy volunteers they had no history of urticaria, rhinitis, asthma, atopic eczema/dermatitis syndrome (AEDS), or other relevant diseases. IgG antibodies to Toxocara canis and Fasciola species were investigated by ELISA method. In a total of 55 patients (mean age: 31.85 ± 8.92), 8 patients (14.5%) were positive for IgG antibodies to Toxocara canis. Among seropositive patients (mean age: 38.62 ± 12.46) 6 patients were female. No patient was positive for Fasciola by ELISA. Six of Toxocara canis seropositive cases were frequently visited or lived in rural areas and had contact with pets. Patients with urticaria, should be tested for Toxocara canis and treated with anthelminthic drugs in endemics areas for toxocariasis.

  13. IgG4-Associated Cholangitis--A Mimic of PSC

    NARCIS (Netherlands)

    Beuers, Ulrich; Hubers, Lowiek M.; Doorenspleet, Marieke; Maillette de Buy Wenniger, Lucas; Klarenbeek, Paul L.; Boonstra, Kirsten; Ponsioen, Cyriel; Rauws, Erik; de Vries, Niek

    2015-01-01

    IgG4-associated cholangitis (IAC) is an inflammatory disorder of the biliary tract representing a major manifestation of IgG4-related disease (IgG4-RD) often with elevation of serum IgG4 levels, infiltration of IgG4+ plasma cells in the affected tissue and good response to immunosuppressive

  14. Fluorescent IgG fusion proteins made in E. coli.

    Science.gov (United States)

    Luria, Yael; Raichlin, Dina; Benhar, Itai

    2012-01-01

    Antibodies are among the most powerful tools in biological and biomedical research and are presently the fastest growing category of new bio-pharmaceutics. The most common format of antibody applied for therapeutic, diagnostic and analytical purposes is the IgG format. For medical applications, recombinant IgGs are made in cultured mammalian cells in a process that is too expensive to be considered for producing antibodies for diagnostic and analytical purposes. Therefore, for such purposes, mouse monoclonal antibodies or polyclonal sera from immunized animals are used. While looking for an easier and more rapid way to prepare full-length IgGs for therapeutic purposes, we recently developed and reported an expression and purification protocol for full-length IgGs, and IgG-based fusion proteins in E. coli, called "Inclonals." By applying the Inclonals technology, we could generate full-length IgGs that are genetically fused to toxins. The aim of the study described herein was to evaluate the possibility of applying the "Inclonals" technology for preparing IgG-fluorophore fusion proteins. We found that IgG fused to the green fluorescent proteins enhanced GFP (EGFP) while maintaining functionality in binding, lost most of its fluorescence during the refolding process. In contrast, we found that green fluorescent Superfolder GFP (SFGFP)-fused IgG and red fluorescent mCherry-fused IgG were functional in antigen binding and maintained fluorescence intensity. In addition, we found that we can link several SFGFPs in tandem to each IgG, with fluorescence intensity increasing accordingly. Fluorescent IgGs made in E. coli may become attractive alternatives to monoclonal or polyclonal fluorescent antibodies derived from animals.

  15. [Study on the production of IgG derived from vaginal epithelial cells and the effect of anti-Candida albicans].

    Science.gov (United States)

    Niu, X X; Li, T; Liu, Z H

    2016-10-25

    Objective: To investigate the function of IgG secreted by vaginal epithelial cells in natural resistance to vulvovaginal candidiasis. Methods: (1)Immunohistochemical method was used to determine the expression of IgG secreted by normal vaginal epithelial cells VK2/E6E7.(2)Samples were divided into three groups by different proportions of VK2/E6E7 cells to Candida albicans ,including Candida albicans: VK2/E6E7 cells were 1∶10, 1∶1[yeast+ cells(1∶10)group and yeast+ cells(1∶1)group]and VK2/E6E7 cells as blank control group. The growth status of 3 groups were observed under inverted microscope after 24 hours. ELISA method was used to detect the production of IgG in 3 groups after 0, 3, 6, 12, 24, 48 hours. Results: (1)Immunohistochemical method showed normal vaginal epithelial cells were polygonal with pale blue nucleus and cytoplasm were distributed by brown granules, which indicated that IgG were strongly positive. While negative control group just had light blue nuclei.(2)Inverted microscope observation represented that control group had a clear outline, strong refraction and large nuclei with cobblestone-like appearance. After yeast+cells(1∶10)group co-cultured for 24 hours, Candida albicans begin to sprout and transformed to hyphae. VK2/E6E7 cells and Candida albicans were close to each other with vacuoles and small black granules in the cytoplasm. The morphology of cells were complete. Yeast+ cells(1∶1)group showed obvious invasion effect of Candida albicans to VK2/E6E7 cells with vigorous growth of hyphae, the decreased number and incomplete morphology of cells. Moreover, the connection of cells were loose. ELISA assay showed that there were statistically significant difference of IgG secretions between the 3 groups after 0, 3, 6, 12, 24, 48 hours( P Candida albicans, secretion of IgG was significantly lower than that in the control group. The statistical difference of IgG secretions between yeast+ cells(1∶10)group and yeast+ cells(1∶1)group

  16. Mathematical Modeling of Avidity Distribution and Estimating General Binding Properties of Transcription Factors from Genome-Wide Binding Profiles.

    Science.gov (United States)

    Kuznetsov, Vladimir A

    2017-01-01

    The shape of the experimental frequency distributions (EFD) of diverse molecular interaction events quantifying genome-wide binding is often skewed to the rare but abundant quantities. Such distributions are systematically deviated from standard power-law functions proposed by scale-free network models suggesting that more explanatory and predictive probabilistic model(s) are needed. Identification of the mechanism-based data-driven statistical distributions that provide an estimation and prediction of binding properties of transcription factors from genome-wide binding profiles is the goal of this analytical survey. Here, we review and develop an analytical framework for modeling, analysis, and prediction of transcription factor (TF) DNA binding properties detected at the genome scale. We introduce a mixture probabilistic model of binding avidity function that includes nonspecific and specific binding events. A method for decomposition of specific and nonspecific TF-DNA binding events is proposed. We show that the Kolmogorov-Waring (KW) probability function (PF), modeling the steady state TF binding-dissociation stochastic process, fits well with the EFD for diverse TF-DNA binding datasets. Furthermore, this distribution predicts total number of TF-DNA binding sites (BSs), estimating specificity and sensitivity as well as other basic statistical features of DNA-TF binding when the experimental datasets are noise-rich and essentially incomplete. The KW distribution fits equally well to TF-DNA binding activity for different TFs including ERE, CREB, STAT1, Nanog, and Oct4. Our analysis reveals that the KW distribution and its generalized form provides the family of power-law-like distributions given in terms of hypergeometric series functions, including standard and generalized Pareto and Waring distributions, providing flexible and common skewed forms of the transcription factor binding site (TFBS) avidity distribution function. We suggest that the skewed binding

  17. Comparative performance of Anigen © FMD NSP Ab ELISA (Korea ...

    African Journals Online (AJOL)

    Comparative performance of Anigen © FMD NSP Ab ELISA (Korea), NSP Priocheck® kit, liquid phase blocking ELISA kits and virus neutralisation tests in the detection of foot and mouth disease virus antibodies in animals sera from Kilifi county, Kenya.

  18. Evaluation of HIV antigen /antibody combination ELISAs for ...

    African Journals Online (AJOL)

    Abstract. Introduction: the aim of this study was to evaluate the performance of Enzygnost HIV Integral II antigen/antibody combination ELISAs in order to formulate HIV ELISA testing algorithms for the Ministry of Health and Social Welfare, Tanzania. Methods: this was a laboratory-based evaluation of Enzygnost HIV Integral ...

  19. DETERMINATION OF HISTAMINE IN FISH USING ELISA TECHNIQUE

    NARCIS (Netherlands)

    KRUGER, C; SEWING, U; STENGEL, G; KEMA, [No Value; WESTERMANN, J; MANZ, B

    1995-01-01

    The analysis of histamine in fish and fish products via competitive ELISA is described. The advantages of this method are easy sample preparation and handling, screening capabilities, and low costs. Automation enables the performance of the assay with higher series of samples. The Histamine-ELISA is

  20. (ELISA) kit for diagnosis copro-antigens of Giardia lamblia

    African Journals Online (AJOL)

    STORAGESEVER

    2010-08-02

    Aug 2, 2010 ... methods based on antigen scanning of parasites such as enzyme linked immunosorbent assay (ELISA), ... samples. To design this method, a pure antibody against parasite as well as an antibody conjugated to a ..... school children in Santiago, Chile by capture ELISA for the detection of fecal Giardia ...

  1. Agricultural production - Phase 2. Indonesia. ELISA for epidemiology of brucellosis

    International Nuclear Information System (INIS)

    Sutherland, S.S.

    1992-01-01

    This document is a travel report of a three-week mission (from October 13 to November 1, 1991) to Indonesia within the framework of ''The implementation of ELISA technology for the sero-diagnosis of important livestock diseases''. The mission evaluated the implementation of ELISA technology at the Regional Laboratories and its role in the surveillance and control of bovine brucellosis

  2. Radioimmunoassay of IgG and IgM rheumatoid factors reacting with human IgG

    International Nuclear Information System (INIS)

    Carson, D.A.; Lawrance, S.; Catalano, M.A.; Vaughan, J.H.; Abraham, G.

    1977-01-01

    Although IgG rheumatoid factor may play a central role in the pathogenesis of rheumatoid arthritis, previously there have been no precise methods for its specific measurement in serum and synovial fluid. This paper describes a solid phase radioimmunoassay for the independent quantification of IgM and IgG rheumatoid factor reacting with the Fc fragment of human IgG. As measured by this assay, serum IgG rheumatoid factor levels differed significantly between patients with seropositive and seronegative rheumatoid arthritis and normal control subjects. In addition, several sera and joint fluids from patients with seropositive rheumatoid arthritis, even without vasculitis, were shown by gel chromatography to have acid-dissociable complexes of IgG rheumatoid factor suggestive of IgG-IgG dimer or trimer formation

  3. Pathomorphological characteristic of IgG4-related diseases

    Directory of Open Access Journals (Sweden)

    O. O. Dyadyk

    2016-08-01

    Full Text Available IgG4-related diseases are a relatively new group of diseases of unknown etiology which are characterized by the development of fibrosis of organs with the presence of big amounts of IgG4-positive plasma-cells in the area of the lesions and increased levels of IgG4 in serum. The organs that may be affected are pancreas, salivary gland, and others, clinical cases of kidney damage are described as well. Renal involvement in IgG4-related diseases most often occurs on the type of tubulointerstitial nephritis, with the further development of acute or chronic kidney injury. The clinic may be represented by the pseudotumor of kidney, renal tissue heterogeneity on the results of CT-studies; acute or chronic renal disease; combination with other organ damage (autoimmune pancreatitis, sclerosing cholangitis, sclerosing lymphoplasmacytic cholecystitis, colitis, sialadenitis, retroperitoneal fibrosis, etc.. Laboratory findings include an increased level of IgG4 in the blood serum, hypocomplementemia, eosinophilia. Histologically, there is interstitial inflammation with many plasma cells, interstitial fibrosis, tubular atrophy, thickening of the tubular basement membrane, some cases are a type of membranous glomerulonephritis. The aim of the study is to identify the patients with IgG4-related diseases with renal impairment and widening the pathological database of such patients with renal impairment to determine the classification criteria of this pathological condition. Materials and methods will include the deceased kidney screening, screening of patients with autoimmune and allergic diseases, nephrological patients screening with the lifetime biopsy (in some cases – repeat biopsy with chronic or acute kidney impairment. There will be clinical and pathological comparison in kidney damage and other diseases with the development of criteria for the classification of lesions in the presence of IgG4-positive substrates and further development of practical

  4. Specific Anti Mumps Antibodies (IgG & IgM in Cerebrospinal Fluid of Mumps Meningoencephalitic Children

    Directory of Open Access Journals (Sweden)

    Noorbakhsh Samileh

    2009-10-01

    Full Text Available Mumps infection is endemic in Iran. Our objective was to evaluate the presence of anti mumps antibodies ( IgM & IgG in cerebrospinal fluid in mumps meningoencephalitic children. A prospective/cross-sectional study was performed in Tehran, Iran (2003 to 2004 and serum anti mumps antibodies (IgM were detected (quantitive; ELISA in meningoencephaltis patients. Specific anti mumps antibodies (IgM & IgG were detected in cerebrospinal fluids of mumps meningoencephalitis cases. 43 meningoencephalitic patients were tested (59.2% male and 40.8% female. The age of patients was 79.96 ± 4.7 month. 23 (78.7% cases had specific mumps IgM in serum. None of cases had IgM antibodies in CSF. Anti mumps IgG antibody was detected in CSF of 7.5% (2/23 cases. We detected lower than expected frequency of local immunity to mumps virus in CSF of our cases. For better serologic diagnosis we recommend more sensitive methods like virus detection (PCR or short-term culture of lymphocytes from cerebrospinal fluid in future studies.

  5. Optimization of ELISA conditions to quantify colorectal cancer antigen-antibody complex protein (GA733-FcK) expressed in transgenic plant.

    Science.gov (United States)

    Ahn, Junsik; Lee, Kyung Jin; Ko, Kisung

    2014-02-01

    The purpose of this study is to optimize ELISA conditions to quantify the colorectal cancer antigen GA733 linked to the Fc antibody fragment fused to KDEL, an ER retention motif (GA733-FcK) expressed in transgenic plant. Variable conditions of capture antibody, blocking buffer, and detection antibody for ELISA were optimized with application of leaf extracts from transgenic plant expressing GA733-FcK. In detection antibody, anti-EpCAM/CD362 IgG recognizing the GA733 did not detect any GA733-FcK whereas anti-human Fc IgG recognizing the human Fc existed in plant leaf extracts. For blocking buffer conditions, 3% BSA buffer clearly blocked the plate, compared to the 5% skim-milk buffer. For capture antibody, monoclonal antibody (MAb) CO17-1A was applied to coat the plate with different amounts (1, 0.5, and 0.25 μg/well). Among the amounts of the capture antibody, 1 and 0.5 μg/well (capture antibody) showed similar absorbance, whereas 0.25 μg/well of the capture antibody showed significantly less absorbance. Taken together, the optimized conditions to quantify plant-derived GA733-FcK were 0.5 μg/well of MAb CO17-1A per well for the capture antibody, 3% BSA for blocking buffer, and anti-human Fc conjugated HRP. To confirm the optimized ELISA conditions, correlation analysis was conducted between the quantified amount of GA733-FcK in ELISA and its protein density values of different leaf samples in Western blot. The co-efficient value R(2) between the ELISA quantified value and protein density was 0.85 (pFcK expression in transgenic plant.

  6. Anti-S. typhi Vi IgG levels in children with and without typhoid vaccinations

    Directory of Open Access Journals (Sweden)

    Sriandayani Sriandayani

    2014-10-01

    Full Text Available Background Typhoid fever is endemic to Indonesia, with an annual incidence of 13/10,000 people. Vaccination has been shown to be an effective method to prevent typhoid fever. Of several vaccine types, the polysaccharide Vi vaccine is the most commonly used typhoid vaccine in developing countries. Results of previous studies remain inconclusive on the necessity of revaccination every 3 years. Objective To compare the mean serum anrioody titers of anti-S. typhi Vi IgG and the proportion of children with protective antibody levels between children with and without typhoid Vi vaccination. Methods We conducted a cross-secrional study at Tuminring District, 11anado from June to September 2012. Data was analyzed using independent T-test and Fisher's test. Serum anti-S. typhi Vi IgG levels were measured by enzyme-linked immunosorbent assay (ELISA method. Results Seventy-six subjects were divided into two groups: 38 children who had received the typhoid Vi vaccination more than 3 years prior to this study and 38 children who never had typhoid vaccinations as a control group. No statistically significant difference in age and gender was found between the two groups. The mean serum anti-Vi IgG level was 0.55 ug/mL (SD 0.58; 95%CI 0.36 to 0.74 in the vaccinated group, significantly higher than that of the control group [0.31 ug/mL (SD 0.12; 950/£1 0.17 to 0.44; P0.0381. The proportion of children with protective antiNi antioody level was higher in the vaccinated group (23.7% than in the control group  (10.5%, howevet; this difference was not statistically significant (P=0.128. Conclusion The mean serum anti-S. typhi Vi IgG antibody level in children who had been vaccinated more than 3 years prior to the study is higher than in children who had never received typhoid vaccinations. Nevertheless, the mean antibody titers are generally non-protective in ooth groups. Also, the proportion of children with protective antibody levels is not significantly

  7. Sandwich ELISA for quantitative detection of human collagen prolyl 4-hydroxylase

    Directory of Open Access Journals (Sweden)

    Myllyharju Johanna

    2010-06-01

    Full Text Available Abstract Background We describe a method for specific, quantitative and quick detection of human collagen prolyl 4-hydroxylase (C-P4H, the key enzyme for collagen prolyl-4 hydroxylation, in crude samples based on a sandwich ELISA principle. The method is relevant to active C-P4H level monitoring during recombinant C-P4H and collagen production in different expression systems. The assay proves to be specific for the active C-P4H α2β2 tetramer due to the use of antibodies against its both subunits. Thus in keeping with the method C-P4H is captured by coupled to an anti-α subunit antibody magnetic beads and an anti-β subunit antibody binds to the PDI/β subunit of the protein. Then the following holoenzyme detection is accomplished by a goat anti-rabbit IgG labeled with alkaline phosphatase which AP catalyzes the reaction of a substrate transformation with fluorescent signal generation. Results We applied an experimental design approach for the optimization of the antibody concentrations used in the sandwich ELISA. The assay sensitivity was 0.1 ng of C-P4H. The method was utilized for the analysis of C-P4H accumulation in crude cell extracts of E. coli overexpressing C-P4H. The sandwich ELISA signals obtained demonstrated a very good correlation with the detected protein activity levels measured with the standard radioactive assay. The developed assay was applied to optimize C-P4H production in E. coli Origami in a system where the C-P4H subunits expression acted under control by different promoters. The experiments performed in a shake flask fed-batch system (EnBase® verified earlier observations that cell density and oxygen supply are critical factors for the use of the inducer anhydrotetracycline and thus for the soluble C-P4H yield. Conclusions Here we show an example of sandwich ELISA usage for quantifying multimeric proteins. The method was developed for monitoring the amount of recombinant C-P4H tetramer in crude E. coli extracts. Due

  8. RYGB progressively increases avidity for a low-energy, artificially sweetened diet in female rats.

    Science.gov (United States)

    Geary, Nori; Bächler, Thomas; Whiting, Lynda; Lutz, Thomas A; Asarian, Lori

    2016-03-01

    Weight re-gain within 2 y after Roux-en-Y gastric bypass (RYGB) is significantly associated with increased intake of and cravings for sweet foods. Here we describe a novel model of this late increase in sweet appetite. Ovariectomized RYGB and Sham-operated rats, with or without estradiol treatment, were maintained on Ensure liquid diet and offered a low-energy, artificially sweetened diet (ASD) 2 h/d. First, we tested rats more than six months after RYGB. ASD meals were larger in RYGB than Sham rats, whereas Ensure meals were smaller. General physical activity increased during ASD meals in RYGB rats, but not during Ensure meals. Second, new rats were adapted to ASD before surgery, and were then offered ASD again during 4-10 wk following surgery. Estradiol-treated RYGB rats lost the most weight and progressively increased ASD intake to >20 g/2 h in wk 9-10 vs. ∼3 g/2 h in Sham rats. Finally, the same rats were then treated with leptin or saline for 8 d. Leptin did not affect body weight, Ensure intake, or activity during meals, but slightly reduced ASD intake in estradiol-treated RYGB rats. Food-anticipatory activity was increased in estradiol-treated RYGB rats during the saline-injection tests. Because increased meal-related physical activity together with larger meals is evidence of hunger in rats, these data suggest that (1) RYGB can increase hunger for a low-energy sweet food in rats and (2) low leptin levels contribute to this hunger, but are not its only cause. This provides a unique rat model for the increased avidity for sweets that is significantly associated with weight recidivism late after RYGB. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Detection of IgM and IgG antibodies to Chlamydophila pneumoniae in pediatric community-acquired lower respiratory tract infections

    Directory of Open Access Journals (Sweden)

    Surinder Kumar

    2011-01-01

    Full Text Available Context: Chlamydophila pneumoniae (C. pneumoniae is an emerging infectious agent with a spectrum of clinical manifestations including lower and upper respiratory tract infections. Aims: To investigate the role of C. pneumoniae in community-acquired lower respiratory tract infections (LRTIs in children using serological tests. Settings and Design: Two hundred children, age 2 months to 12 years, hospitalized for community-acquired LRTIs were investigated for C. pneumoniae etiology. Materials and Methods: We investigated 200 children hospitalized for community-acquired LRTIs, using ELISA for detecting anti-C. pneumoniae IgM and IgG antibodies. The demographic, clinical and radiological findings for C. pneumoniae antibody positive and C. pneumoniae antibody negative cases were compared. Statistical Analysis Used: Data analysis was performed by Chi-square test and Fisher′s exact tests using Epi Info (2002. Results: Clinical and radiological findings in both the groups were comparable. Serological evidence of C. pneumoniae infection was observed in 12 (6% patients; specific IgM antibodies were detected in 11 (91.67%; specific IgG antibodies in 1 (8.33% patients, while 4-fold rise in C. pneumoniae IgG antibody titers were noted in none of the patients. Conclusions: C. pneumoniae has a role in community-acquired LRTIs, even in children aged < 5 years. Serological detection using ELISA would enable pediatricians in better management of C. pneumoniae infections.

  10. Plasmonic ELISA for the ultrasensitive detection of Treponema pallidum.

    Science.gov (United States)

    Nie, Xin-Min; Huang, Rong; Dong, Cai-Xia; Tang, Li-Juan; Gui, Rong; Jiang, Jian-Hui

    2014-08-15

    In this report, we have developed a plasmonic ELISA strategy for the detection of syphilis. Plasmonic ELISA is an enzyme-linked immunoassay combined with enzyme-mediated surface plasmon resonance (SPR) of gold nanoparticles (AuNPs). Immune response of the Treponema pallidum (T. pallidum) antibodies triggers the acetylcholinesterase-catalyzed hydrolysis of acetylthiocholine to produce abundant thiocholine. The positive charged thiol, in turn, alters the surface charge distribution the AuNPs and leads to the agglomeration of the AuNPs. The induced strong localized SPR effect of the agglomerate AuNPs can, thus, allow the quantitative assay of T. pallidum antibodies due to the remarkable color and absorption spectral response changes of the reaction system. The plasmonic ELISA exhibited a quasilinear response to the logarithmic T. pallidum antibody concentrations in the range of 1pg/mL-10ng/mL with a detection limit of 0.98pg/mL. Such a low detection limit was 1000-fold improvements in sensitivity over a conventional ELISA. The results of plasmonic ELISA in syphilis assays of serum specimens from 60 patients agreed with those obtained using a conventional ELISA method. The plasmonic ELISA has characteristics (analyte specific, cost-effective, ease of automatic, low limit of detection) that provide potential for diagnosis and therapeutic monitoring of syphilis. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Serological blind spots for variants of human IgG3 and IgG4 by a commonly used anti-immunoglobulin reagent.

    Science.gov (United States)

    Howie, Heather L; Delaney, Meghan; Wang, Xiaohong; Er, Lay See; Vidarsson, Gestur; Stegmann, Tamara C; Kapp, Linda; Lebedev, Jenna N; Wu, Yanyun; AuBuchon, James P; Zimring, James C

    2016-12-01

    Human immunoglobulin G (IgG) includes four different subtypes (IgG1, IgG2, IgG3, and IgG4), and it is also now appreciated that there are genetic variations within IgG subtypes (called isoallotypes). Twenty-nine different isoallotypes have been described, with 7, 4, 15, and 3 isoallotypes described for IgG1, IgG2, IgG3, and IgG4, respectively. The reactivity of anti-IgG with different isoallotypes has not been characterized. A novel monoclonal anti-K antibody (PugetSound Monoclonal Antibody 1 [PUMA1]) was isolated and sequenced, and a panel of PUMA1 variants was expressed, consisting of the 29 known IgG isoallotypes. The resulting panel of antibodies was preincubated with K-positive red blood cells (RBCs) and then subjected to testing with currently approved anti-IgG by flow cytometry, solid phase systems, gel cards, and tube testing. A US Food and Drug Administration (FDA)-approved monoclonal anti-IgG (gamma-clone) failed to recognize 2 of 15 IgG3 isoallotypes (IgG3-03 and IgG3-13) and 3 of 3 IgG4 isoallotypes (IgG4-01, IgG4-02, and IgG4-03). In contrast, an FDA-approved rabbit polyclonal anti-IgG recognized each of the known human IgG isoallotypes. These findings demonstrate "blind spots" in isoalloantibody detection by a monoclonal anti-IgG. If a patient has anti-RBC antibodies predominantly of an IgG3 subtype (the IgG3-03 and/or IgG3-13 variety), then it is possible that a clinically significant alloantibody would be missed. IgG-03 and IgG-13 have an estimated frequency of 1% to 3% in Caucasian populations and 20% to 30% in certain African populations. Nonreactivity with IgG4 is a known characteristic of this monoclonal anti-IgG, but IgG4 isoallotypes have not been previously reported. © 2016 AABB.

  12. Usefulness of in-house obtained recombinant proteins Yop of Yersinia enterocolitica as highly specific antigens in ELISA and recom-dot performed in the serodiagnosis of yersiniosis.

    Science.gov (United States)

    Rastawicki, Waldemar; Smietafiska, Karolina; Chrost, Anna; Wolkowicz, Tomasz; Rokosz-Chudziak, Natalia

    Proper analysis of the human immune response is crucial in the laboratory diagnosis of many bacterial infections-The current serological diagnosis of yersiniosis often is carried out using ELISA with native antigens. However, recombinant proteins increase the specificity of the serological assays, particularly in patients with chronic, non- specific infections. The aim of the present study was to evaluate the usefulness of in-house obtained recombinant proteins Yop of Yersinia enterocolitica as highly specific antigens in ELISA and recom-dot performed in the serodiagnosis of yersiniosis. Recombinant YopD, YopB, YopE and V-Ag proteins of Y enterocolitica were expressing in E. coli BL21 (DE3) using the pET-30 Ek/LIC expression vector (Novagen). Purification was accomplished by immobilized metal (Ni2) affinity column chromatography (His-trap). The proteins were used as antigens in standard ELISA and recom-dot assay, which was performed on nitrocellulose strips. The study population, used for characterization of the humoral immune response to the recombinant proteins, consisted of 74 patients suspected for Y enterocolitica infection and 41 clinically healthy blood donors. Some of the results obtained by ELISA and recom-dot were compared with results obtained by commercial western-blot Yersinia (Virotech). In the group of patients suspected for yersiniosis in clinical investigation the most positive results were obtained in ELISA with the recombinant protein YopD (IgA respectively 25 (42.4%), IgG 41 (69.5%), IgM 24 (40.7%). The percentage ofpositive results in the group of blood donors did not exceed 10.0% in IgG and 5.0% in IgA/IgM classes of immunoglobulin. The results obtained in the recom-dot assay showed that among 74 tested serum samples obtained from individuals suspected of yersiniosis the most common IgA, IgG and IgM antibodies were found for recombinant protein YopD (respectively IgG in 60.8%, IgA in 37.8% and IgM in 33.8% of serum samples). IgG antibodies to

  13. Food allergen-specific serum IgG and IgE before and after elimination diets in allergic dogs.

    Science.gov (United States)

    Zimmer, Anja; Bexley, Jennifer; Halliwell, Richard E W; Mueller, Ralf S

    2011-12-15

    Serum food allergen-specific antibody testing is widely offered to identify suitable ingredients for diets to diagnose adverse food reaction (AFR) in dogs with allergic skin disease. Antibody concentrations in blood samples obtained during an unsuccessful diet to help in the choice of diet changes may be influenced by the previous diet. The objective of this paper was to measure food antigen-specific IgE and IgG for the most commonly used 16 food antigens before and after an elimination diet. Levels of food-specific serum IgE and IgG antibodies were measured by enzyme-linked immunosorbent assay (ELISA). Dogs had detectable IgE antibodies to beef, pork, lamb and cows' milk; and detectable IgG antibodies to beef, pork, lamb, cows' milk, chicken and turkey. Of 19 dogs with complete data sets, 14 dogs showed clear improvement during diet and in 7 dogs AFR could be diagnosed by deterioration on rechallenge and subsequent improvement on refeeding the diet. Serum was obtained before and 6-8 weeks after beginning such a diet. There was no significant difference in pre- and post-diet levels for any of the individual allergens nor for the total IgE and IgG concentrations of all antigens (P=0.55 and P=0.53 respectively). In these 19 dogs in which an elimination diet was used for the diagnosis of food allergy and in which 14 were probably food allergic and 7 were proven food allergic there were no significant differences in food-specific antibodies before and after an elimination diet of 6-8 weeks. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Assessment of humoral immunity to Eimeria tenella sporozoites in chickens by ELISA

    Directory of Open Access Journals (Sweden)

    S. Saravanan

    2014-07-01

    Full Text Available Aim: To assess the humoral immune response of Eimeria tenella sporozoites in broiler chickens by a developed enzyme linked immunosorbent assay (ELISA and the efficacy in terms of bodyweight, lesion score and oocysts excretion in immunized broilers. Materials and Methods: Purified live E. tenella sporozoites were administered subcutaneously in neck region of broiler chickens in the early life (first week at different concentrations. The potency of the sporozoite vaccine as assessed by IgG levels and the performance in immunized broilers as assessed by body weight, lesion score and oocysts excretion in faeces after challenge with 10, 000 live E. tenella oocysts at 49 days of age were evaluated. Results: The chickens of group (T4 immunized with 20 µg of antigen on day 6 showed an increase in IgG levels (0.161±0.004 two weeks post immunization (PI peaking (0.399± 0.016 at 5 weeks PI. The mean weekly weight gain (g after challenge, at 56 days of age was high in T4 (148±4.751 g with a low mean lesion score (2.5±0.22 and mean oocyst output (x103 oocytes per gram (OPG in faeces (100.3± 45.72 when compared to unimmunised infected controls. Conclusion: An early but partial immune response against caecal coccidiosis could be achieved by immunization with E. tenella specific sporozoites in chickens of less than a week old. Moreover, the performance of immunized chickens as indicated by weight gain, lesion score and oocyst output was found to be superior to the unimmunized infected controls.

  15. Localized IgG4-related Cholecystitis Mimicking Gallbladder Cancer.

    Science.gov (United States)

    Inoue, Tadahisa; Okumura, Fumihiro; Mizushima, Takashi; Nishie, Hirotada; Iwasaki, Hiroyasu; Anbe, Kaiki; Ozeki, Takanori; Kachi, Kenta; Fukusada, Shigeki; Suzuki, Yuta; Watanabe, Kazuko; Sano, Hitoshi

    2015-01-01

    We encountered a case of localized IgG4-cholecystitis mimicking gallbladder cancer with focal/segmental type1 autoimmune pancreatitis (AIP). In this case, we were unable to exclude a diagnosis of gallbladder cancer and thus performed radical cholecystectomy. Type1 AIP is often associated with gallbladder lesions, accompanied by generally diffuse, circumferential thickening of the gallbladder wall. Although localized IgG4-related cholecystitis is extremely rare, differentiating this condition from gallbladder cancer is often very difficult.

  16. Serologic cross-reactivity of human IgM and IgG antibodies to five species of Ebola virus.

    Directory of Open Access Journals (Sweden)

    Adam Macneil

    2011-06-01

    Full Text Available Five species of Ebola virus (EBOV have been identified, with nucleotide differences of 30-45% between species. Four of these species have been shown to cause Ebola hemorrhagic fever (EHF in humans and a fifth species (Reston ebolavirus is capable of causing a similar disease in non-human primates. While examining potential serologic cross-reactivity between EBOV species is important for diagnostic assays as well as putative vaccines, the nature of cross-reactive antibodies following EBOV infection has not been thoroughly characterized. In order to examine cross-reactivity of human serologic responses to EBOV, we developed antigen preparations for all five EBOV species, and compared serologic responses by IgM capture and IgG enzyme-linked immunosorbent assay (ELISA in groups of convalescent diagnostic sera from outbreaks in Kikwit, Democratic Republic of Congo (n=24, Gulu, Uganda (n=20, Bundibugyo, Uganda (n=33, and the Philippines (n=18, which represent outbreaks due to four different EBOV species. For groups of samples from Kikwit, Gulu, and Bundibugyo, some limited IgM cross-reactivity was noted between heterologous sera-antigen pairs, however, IgM responses were largely stronger against autologous antigen. In some instances IgG responses were higher to autologous antigen than heterologous antigen, however, in contrast to IgM responses, we observed strong cross-reactive IgG antibody responses to heterologous antigens among all sets of samples. Finally, we examined autologous IgM and IgG antibody levels, relative to time following EHF onset, and observed early peaking and declining IgM antibody levels (by 80 days and early development and persistence of IgG antibodies among all samples, implying a consistent pattern of antibody kinetics, regardless of EBOV species. Our findings demonstrate limited cross-reactivity of IgM antibodies to EBOV, however, the stronger tendency for cross-reactive IgG antibody responses can largely circumvent

  17. Radiation Therapy to the Primary and Postinduction Chemotherapy MIBG-Avid Sites in High-Risk Neuroblastoma

    Energy Technology Data Exchange (ETDEWEB)

    Mazloom, Ali; Louis, Chrystal U.; Nuchtern, Jed; Kim, Eugene; Russell, Heidi; Allen-Rhoades, Wendy; Krance, Robert; Paulino, Arnold C., E-mail: apaulino@mdanderson.org

    2014-11-15

    Purpose: Although it is generally accepted that consolidation therapy for neuroblastoma includes irradiation of the primary site and any remaining metaiodobenzylguanidine (MIBG)-avid metastatic sites, limited information has been published regarding the efficacy of this approach. Methods and Materials: Thirty patients with high-risk neuroblastoma were treated at 1 radiation therapy (RT) department after receiving 5 cycles of induction chemotherapy and resection. All patients had at least a partial response after induction therapy, based upon international neuroblastoma response criteria. The primary sites were treated with 24 to 30 Gy whereas the MIBG-avid metastatic sites were treated with 24 Gy. RT was followed by high-dose chemotherapy with autologous stem cell rescue and 6 months of cis-retinoic acid. Results: The 5-year progression-free survival (PFS) and overall survival (OS) rates were 48% and 59%, respectively. The 5-year locoregional control at the primary site was 84%. There were no differences in locoregional control according to degree of primary surgical resection. The 5-year local control rate for metastatic sites was 74%. The 5-year PFS rates for patients with 0, 1, 2, and >3 postinduction MIBG sites were 66%, 57%, 20%, and 0% (P<.0001), respectively, whereas 5-year OS rates were 80%, 57%, 50%, and 0%, respectively (P<.0001). Conclusions: RT to the primary site and postinduction MIBG-positive metastatic sites was associated with 84% and 74% local control, respectively. The number of MIBG-avid sites present after induction chemotherapy and surgery was predictive of progression-free and overall survival.

  18. Favourable outcomes of {sup 177}Lu-octreotate peptide receptor chemoradionuclide therapy in patients with FDG-avid neuroendocrine tumours

    Energy Technology Data Exchange (ETDEWEB)

    Kashyap, Raghava [Peter MacCallum Cancer Center, Centre for Cancer Imaging, Melbourne (Australia); Hofman, Michael S.; Kong, Grace; Akhurst, Timothy; Eu, Peter [Peter MacCallum Cancer Center, Centre for Cancer Imaging, Melbourne (Australia); Peter MacCallum Cancer Centre, Neuroendocrine Tumour Service, Melbourne (Australia); Michael, Michael [University of Medicine, Department of Medicine, Melbourne (Australia); Peter MacCallum Cancer Centre, Division of Cancer Medicine, Melbourne (Australia); University of Melbourne, The Sir Peter MacCallum Department of Oncology, Melbourne (Australia); Zannino, Diana [Peter MacCallum Cancer Centre, Biostatistics and Clinical Trials, Melbourne (Australia); Hicks, Rodney J. [Peter MacCallum Cancer Center, Centre for Cancer Imaging, Melbourne (Australia); Peter MacCallum Cancer Centre, Neuroendocrine Tumour Service, Melbourne (Australia); University of Melbourne, The Sir Peter MacCallum Department of Oncology, Melbourne (Australia)

    2014-09-11

    Increased glycolytic activity on FDG PET/CT defines a subgroup of patients with metastatic gastroenteropancreatic neuroendocrine tumour (NET) with a poor prognosis. A limited range of systemic treatment options exist for more aggressive NET. The role of peptide receptor chemoradionuclide therapy (PRCRT) in such patients is, however, unclear. This retrospective study assessed the outcomes of patients with FDG-avid NET treated with PRCRT. Clinical, biochemical and imaging response was assessed after completion of induction treatment of PRCRT with 5-fluorouracil in 52 patients selected for treatment on the basis of somatostatin-receptor imaging without spatially discordant FDG-avid disease. Of the cohort, 67 % had received prior chemotherapy. Overall survival (OS) and progression-free survival (PFS) were also analysed. PRCRT was well tolerated with negligible grade 3/4 toxicities. After a median follow-up period of 36 months, the median OS was not achieved with a median PFS of 48 months. At 3 months after completion of PRCRT 2 % of patients showed a complete anatomical response, 28 % a partial response, 68 % stable disease, and only 2 % progression. On FDG PET/CT, 27 % achieved a complete metabolic response during the follow-up period. A biochemical response (>25 % fall in chromogranin-A levels) was seen in 45 %. PRCRT is an effective treatment in patients with FDG-avid NET, even in patients who have failed conventional therapies. Given apparently higher response rates than with alternative therapeutic options and low toxicity, further research is needed to establish whether PRCRT should be used as a first-line treatment modality in this patient population. (orig.)

  19. "Auto-anti-IgE": naturally occurring IgG anti-IgE antibodies may inhibit allergen-induced basophil activation.

    Science.gov (United States)

    Chan, Yih-Chih; Ramadani, Faruk; Santos, Alexandra F; Pillai, Prathap; Ohm-Laursen, Line; Harper, Clare E; Fang, Cailong; Dodev, Tihomir S; Wu, Shih-Ying; Ying, Sun; Corrigan, Christopher J; Gould, Hannah J

    2014-12-01

    Naturally occurring IgE-specific IgG autoantibodies have been identified in patients with asthma and other diseases, but their spectrum of functions is poorly understood. Address the hypothesis that: (i) IgG anti-IgE autoantibodies are detectable in the serum of all subjects but elevated in asthmatic patients regardless of atopic status as compared with controls; (ii) some activate IgE-sensitized basophils; and (iii) some inhibit allergen-induced basophil activation. IgE-specific IgG autoantibodies were detected and quantified in sera using ELISA. Sera were examined for their ability to activate IgE-sensitized human blood basophils in the presence and absence of allergen using a basophil activation test, and to inhibit allergen binding to specific IgE on a rat basophilic cell line stably expressing human FcεRI. IgG autoantibodies binding to both free and FcεRI-bound IgE were detected in patients with atopic and non-atopic asthma, as well as controls. While some were able to activate IgE-sensitised basophils, others inhibited allergen-induced basophil activation, at least partly by inhibiting binding of IgE to specific allergen. Naturally occurring IgG anti-IgE autoantibodies may inhibit, as well as induce, basophil activation. They act in a manner distinct from therapeutic IgG anti-IgE antibodies such as omalizumab. They may at least partly explain why atopic subjects who make allergen-specific IgE never develop clinical symptoms, and why omalizumab therapy is of variable clinical benefit in severe atopic asthma. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Evaluation of anti-Schistosoma mansoni igG antibodies in patients with chronic schistosomiasis mansoni before and after specific treatment Avaliação da presença de anticorpos IgG anti-Schistosoma mansoni no soro de pacientes com esquistossomose mansônica crônica, antes e após tratamento específico

    Directory of Open Access Journals (Sweden)

    Célia Maria V. VENDRAME

    2001-06-01

    Full Text Available The circumoval precipitin test (COPT, enzyme-linked immunosorbent assay (ELISA and the immunoblotting anti-adult worm antigen (AWA and soluble egg antigen (SEA tests were applied to 17 chronically schistosome-infected patients for the detection of anti-Schistosoma mansoni antibodies before and on four occasions after oxamniquine administration over a period of six months. Compared to a control group, schistosomiasis patients showed high levels of IgG antibodies in AWA and SEA-ELISA. A decrease in IgG levels was observed six months after treatment, although negative reactions were not obtained. Significant decreases in IgG1, IgG3 and, mainly, IgG4, but not anti-SEA IgG2 levels were observed six months after treatment, again without negativity. Analysis of anti-AWA IgG antibodies by immunoblotting before treatment showed a 31 kDa strand in 14 patients (82% which disappeared in three cases up to six months after treatment; furthermore, anti-SEA IgG antibodies showed the same band in nine patients (53% before treatment, which disappeared in only four cases up to six months after treatment.Em 17 pacientes com infecção crônica por Schistosoma mansoni utilizaram-se os testes de reação periovular, imunoenzimático (ELISA e imunoblotting, empregando-se antígenos obtidos a partir de vermes adultos (AWA ou de ovos de S. mansoni (SEA, para detecção de anticorpos anti-S. mansoni, antes e em quatro ocasiões após tratamento com oxamniquine. Quando cotejados a grupo controle os pacientes esquistossomóticos revelaram altos níveis séricos de anticorpos IgG nos testes ELISA (anti-AWA e anti-SEA, não se observando, porém, negativação até seis meses após tratamento específico. Encontrou-se, entretanto, decréscimo significativo, sem negativação, dos níveis de IgG1, IgG3 e, principalmente, IgG4, quando se utilizou antígeno solúvel obtido a partir de ovos de S. mansoni (SEA, seis meses após administração de oxamniquine. O mesmo não foi

  1. Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA and indirect ELISA Detecção de anticorpos anti-Toxoplasma gondii por meio das técnicas de Imunofluorescência Indireta e ELISA Indireto em primatas experimentalmente e naturalmente infectados

    Directory of Open Access Journals (Sweden)

    Andréa Bouer

    2010-03-01

    Full Text Available The Indirect Fluorescence Assay (IFA and the indirect ELISA were comparatively used to detect IgG and IgM antibodies for Toxoplasma gondii in experimentally and naturally infected primates. In the experimentally infected group, antibodies of diagnostic value were detected at day 9 post-infection (PI with the IFA (IgG and IgM and with IgG-ELISA. IgM-ELISA detected antibodies for T. gondii starting at day 3 PI until the end of the experiment (102 days PI. Of the 209 naturally infected sera tested, from many zoos of State of Sao Paulo, 64.59 and 67.94% were positive in the IgG-IFA test and IgG-ELISA respectively. IgM-ELISA test detected seropositivity in 52.63% of the sera although IgM-IFA test detected it in only in 0.96% of the samples. The differential toxoplasmosis diagnosis was accomplished with Neospora caninum by IFA, observing 61 (29.2% seropositive animals for this parasite and 149 (70.8% negative. Sixty animals were positive for both T. gondii and N. caninum. Pneumonia, splenomegaly, and intestinal ulcers were macroscopically observed. Unremarkable interstitial pneumonia, enteritis, colitis, splenitis, and glomerulitis were microscopically observed. The immunohistochemical stain could not detect the presence of T. gondii in the tissues of the animals infected experimentally.Detectou-se anticorpos das classes IgG e IgM anti-Toxoplasma gondii em primatas experimentalmente e naturalmente infectados, utilizando-se como técnicas comparativas a RIFI e o ELISA-teste. No grupo dos primatas experimentalmente infectados, anticorpos de valor diagnóstico foram detectados a partir do 9º dia de infecção tanto na RIFI (IgG e IgM como no ELISA-IgG. O ELISA IgM detectou anticorpos a partir do 3º dia de infecção até o final do experimento (102 dias pós-infecção. Dos 209 soros dos primatas naturalmente infectados, de diversos zoológicos do Estado de São Paulo, 64,59 e 67,94% mostraram-se positivos na RIFI-IgG e no ELISA-IgG, respectivamente. O

  2. Detection of Fasciola gigantica antibodies using Pourquier ELISA kit ...

    African Journals Online (AJOL)

    ELISA) screening kit for Fasciola antibodies was conducted in breeding herds in two Local Government Areas of Adamawa state. The objectives were to determine the presence of Fasciola gigantica antibodies as a way of demonstrating the use ...

  3. Grantee Spotlight: Elisa Rodriguez, Ph.D., M.S.

    Science.gov (United States)

    Dr. Elisa M. Rodriguez tests the feasibility of community-based participatory research approaches to engaging Hispanics, African Americans, and the medically underserved in the Buffalo, NY area in biospecimen donation for cancer research.

  4. Reliable identification of human albumin in ancient bone using ELISA and monoclonal antibodies.

    Science.gov (United States)

    Cattaneo, C; Gelsthorpe, K; Phillips, P; Sokol, R J

    1992-03-01

    In order to help reconstruct ancient dietary, domestic, and ritual behaviour, a method was developed to identify the blood protein albumin in ancient skeletal material. This was an inhibition enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody of IgG class against human albumin. With fresh material, the technique gave consistent and specific results and could detect as little as 10 ng of albumin. Extracts of bone from the English Civil War (A.D. 1644), Mediaeval (A.D. 1100-1400), Early Saxon (A.D. 450-600), Roman (A.D. 100-200), Iron Age (ca 400 B.C.) and Bronze Age (2200-1700 B.C. cal.) periods were then tested, samples of fresh human and animal bone being included as positive and negative controls, respectively. Albumin was demonstrated in human bone from all periods; there was no evidence of cross-reactivity with animal material. Detection seemed to depend largely on amount of sample and chronological age; other factors, such as physical integrity of the specimen and soil characteristics, appeared to be less important. Preliminary studies of other ancient skeletal remains showed that animal species could be readily identified and that albumin was probably still detectable in cremated material. It is concluded that our method provides a tool specific and sensitive enough for the reliable identification of the species-origin of small fragments of bone (and possibly of blood stains) and will thus allow insight into past behaviour patterns. ELISA may also be suitable for identifying other molecules (such as HLA and ABO) which would help determine racial affiliation and disease predisposition among ancient populations.

  5. Development of an LPS-based ELISA for diagnosis of Yersinia enterocolitica O:3 infections in Danish patients: a follow-up study.

    Science.gov (United States)

    Dalby, Tine; Rasmussen, Eva; Schiellerup, Peter; Krogfelt, Karen Angeliki

    2017-05-25

    The bacterium Yersinia enterocolitica causes gastroenteritis in humans. The study aimed to develop a diagnostic enzyme-linked immunosorbent assay (ELISA) for detection of Yersinia enterocolitica O:3 LPS antibodies in sera from Danish patients with suspected Yersinia enterocolitica O:3 gastrointestinal infection. As a part of this, antibody decay profiles after culture confirmed Yersinia enteritis were studied. An ELISA using Yersinia enterocolitica O:3 LPS as the coating antigen was developed for measuring IgA, IgG and IgM specific antibodies. A longitudinal collection of 220 sera drawn between 20 and 1053 days after onset of symptoms from 85 adult Danish patients with verified Yersinia enteritis were examined. A control group of 100 sera from healthy Danish blood-donors were analysed in order to determine the cut-off for interpretation of results. Serum samples from 62 out of 81 patients who delivered either the first or the second sample were found positive for specific antibodies against Yersinia enterocolitica O:3 LPS (77%). For samples collected within 60 days after onset of symptoms (n = 48) sensitivities of 58%, 42% and 79% for IgA, IgG and IgM antibodies were found. A sensitivity of 81% was found for these samples when using the definition of a positive result in either IgA, IgG or IgM as a combined positive. All samples received up to 36 days after onset of symptoms (n = 10) were found to be positive using this definition. For the period 61 to 90 days after onset of symptoms (n = 32), a combined sensitivity of 63% was found. The antibody levels as well as decay profiles for the three different immunoglobulin classes for the individual patients exhibited a large degree of variation. Using a definition of positive as a positive result for either IgA, IgG or IgM antibodies, a diagnostic sensitivity of 81% was achieved for samples received within 60 days after onset of symptoms. In particular, the levels of specific IgM antibodies were elevated. In

  6. Anti-CagA IgG Antibody is Independent from Helicobacter pylori vacA and cagA Genotypes

    Directory of Open Access Journals (Sweden)

    Hashem Fakhre Yaseri

    2015-12-01

    Full Text Available Background: Helicobacter pylori strains have two classical virulence genes, the cytotoxinassociated A (cagA gene and the vacuolating cytotoxin A (vacA gene, which are located in thecag pathogenicity island (cagPAI. Serum immunoglobulin G (IgG antibodies to H. pylori,especially, the CagA antigen may be a reliable marker for selection of dyspeptic patients for upperendoscopy.Methods: Serum sample of 129 dyspeptic patients with positive H. pylori, were tested for serumIgG Anti-CagA antibody by ELISA. The presence of the cagA and vacA genotypes weredetermined using polymerase chain reaction (PCR on biopsy samples taken via endoscopy.Results: Positive serum IgG anti-CagA antibodies in patients with cagA+/vacA+ and cagA+/vacA- genotypes were 22/23 (95.6% and 18/19 (94.7%, respectively. In addition, serum IgG anti-CagAantibodies in patients with cagA-/vacA+ and cagA-/vacA- genotypes were 22/47 (46.8% and 33/40(82.5%, respectively.Conclusions: It can be concluded that the serum IgG anti-CagA antibody alone could selectpatients with dyspepsia following upper endoscopy. The assessment of vacuolating cytotoxinactivity of H. Pylori is, therefore, not required, even when vacA gene is positive. This hypothesisneeds to be studied in a large number of patients with dyspepsia.

  7. A quantitative ELISA for monitoring the secretion of ZZ-fusion proteins using SpA domain as immunodetection reporter system.

    Science.gov (United States)

    Mergulhão, F J; Monteiro, G A; Cabral, J M; Taipa, M A

    2001-11-01

    A sandwich-type enzyme-linked immunosorbent assay (ELISA) was established for monitoring the secretion of ZZ-fusion proteins. Two antibodies, a monoclonal mouse anti-human proinsulin and a rabbit anti-bovine IgG (strongly binding to the ZZ-domain), were used to quantify the secretion of recombinant human ZZ-proinsulin to the growth medium of Escherichia coli cultures. The method here reported conjugates the advantages of sandwich-type ELISA assays, namely, high sensitivity, specificity, and throughput, with the possibility of quantifying small protein molecules (e.g., peptides). A further advantage of gene fusion techniques integrating both downstream processing and product detection and quantitation is highlighted. The method is capable of detecting levels of 0.05 ng of ZZ-proinsulin.

  8. High prevalence of human anti-bovine IgG antibodies as the major cause of false positive reactions in two-site immunoassays based on monoclonal antibodies

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Koch, Claus; Jensen, Charlotte H

    2004-01-01

    were purified by protein G affinity chromatography from culture supernatant containing 10% (v/v) fetal calf serum (FCS). Human anti-animal IgG (bovine, mouse, horse, and swine) antibodies and human anti-bovine serum albumin antibodies were measured using an ELISA design, with direct bridging...... of the solid phase and biotinylated antigens. The false positive reactions were abolished by addition of 1% (v/v) bovine serum to the dilution buffer (DB). Human anti-bovine IgG antibodies (HABIA) were detected in 99 out of 104 sera from blood donors (50 females; 54 males). HABIA levels in male sera (n = 54...... positively correlated to human anti-bovine serum albumin antibodies in the donor sera (r = 0.639; p

  9. Characteristic tubulointerstitial nephritis in IgG4-related disease.

    Science.gov (United States)

    Yamaguchi, Yutaka; Kanetsuna, Yukiko; Honda, Kazuho; Yamanaka, Nobuaki; Kawano, Mitsuhiro; Nagata, Michio

    2012-04-01

    Nephropathy associated with IgG4-related disease is characterized by tubulointerstitial nephritis. To better identify its pathology, the present study analyzed clinicopathologic features of IgG4-related tubulointerstitial nephritis cases from across Japan. Sixteen cases were identified as IgG4-related nephropathy using the criterion of high serum IgG4 levels (>135 mg/dL) with abnormal kidney computed tomography or elevated serum creatinine levels. Male predominance (75%) and advanced age (average, 62.0 years) were noted. Eight cases displayed no autoimmune pancreatitis. Renal computed tomography abnormalities were found in 12 of 13 cases examined. Renal dysfunction was found in 15 of 16 cases at biopsy. Distinctive features of tubulointerstitial lesions included (1) well-demarcated borders between involved and uninvolved areas; (2) involvement of the cortex and medulla, often extending beyond the renal capsule and with occasional extension to retroperitoneal fibrosis; (3) interstitial inflammatory cells comprising predominantly plasma cells and lymphocytes, with a high prevalence of IgG4-positive cells often admixed with fibrosis; (4) peculiar features of interstitial fibrosis resembling a "bird's-eye" pattern comprising fibrosis among inter-plasma cell spaces; and (5) deposits visible by light and immunofluorescent microscopy in the tubular basement membrane, Bowman capsule, and interstitium that are restricted to the involved portion, sparing normal parts. Ultrastructural analysis revealed the presence of myofibroblasts with intracellular/pericellular collagen accompanied by plasma cell accumulation from an early stage. Histology could not discriminate between IgG4-related tubulointerstitial nephritis with and without autoimmune pancreatitis. In conclusion, the distinctive histologic features of IgG4-related tubulointerstitial nephritis can facilitate the differential diagnosis of tubulointerstitial nephritis, even without autoimmune pancreatitis or an abnormal

  10. Differential expression profiling of circulation microRNAs in PTC patients with non-131I and 131I-avid lungs metastases: a pilot study

    International Nuclear Information System (INIS)

    Qiu, Zhong-Ling; Shen, Chen-Tian; Song, Hong-Jun; Wei, Wei-Jun; Luo, Quan-Yong

    2015-01-01

    Introduction: Loss of the ability to concentrate 131 I is one of the important causes of radioiodine-refractory disease in papillary thyroid carcinoma (PTC). Recent advantages of serum microRNAs (miRNAs) open a new realm of possibilities for noninvasive diagnosis and prognosis of many cancers. The aim of the current study was to identify differential expression profiling of circulation miRNAs in PTC patients with non- 131 I and 131 I-avid lungs metastases. Methods: The expressions of miRNAs were examined using miRNA microarray chip. The most significantly changed miRNAs from microarray were verified by using qRT-PCR. The potential miRNAs regulating target genes and their preliminary biological functions were forecasted by Bioinformatic analysis. Results: Compared to 131 I-avid lung metastases, 13 kinds of significantly differential serum miRNAs including 5 upregulated miRNAs (miR-1249, miR-106a, miR-503, miR-34c-5p, miR-1281) and 8 downregulated miRNAs (miR-1915, miR-2861, miR-3196, miR-500, miR-572, miR-33b, miR-554, miR-18a) in PTC patients with non- 131 I-avid lung metastases were identified. Bioinformatic analysis demonstrated that miR-106a was the core miRNA regulating 193 genes in the network. The results of validation confirmed the up-regulation of miR-106a in non- 131 I-avid lungs metastatic PTC patients. Conclusion: Differentially expressed serum miRNA profiles between PTC patients with non- 131 I and 131 I-avid lungs metastases were analyzed. These findings in our present study could represent new clues for the diagnostic and therapeutic strategy in PTC patients with non- 131 I-avid metastatic disease

  11. Study on immobilizations of ovine anti-human IgG and MCAb against EHF on radiation-modified silicone films

    International Nuclear Information System (INIS)

    Guo Jinhui; Ha Hongfei; Zhang Yuhua

    1990-01-01

    Films of silicone (silastic) were grafted by monomer acrylamide vis γ-radiation technology and then the ovine anti-human IgG, Epidemic hemorrhagic fever (EHF)-MCAb were immobilized on the silastic-AAM films with different grafting yields passthrough associate reactions. Measruements of relationships between grafting yields. Contents of immobilized antibodies and immunoactivities for immobilized silastic-AAM films were performed by using 125 I method ELISA method was used to measure the immunoactivities for the immobilized monoantibody. The results showed that the antibodies used can be immobilized on radiation-grafted silicone films and this immobilization method has its potential significance in clinical practice

  12. Diagnostic Value of PCR and ELISA for Chlamydia trachomatis in a Group of Asymptomatic and Symptomatic Women in Isfahan, Iran

    Directory of Open Access Journals (Sweden)

    Anahita Jenab

    2009-01-01

    Full Text Available Background: Chlamydia trachomatis infections are the most prevalent sexually transmittedbacterial infections (STI in the world that lead to a cause of tubal factor infertility in women. Theaim of this study is to determine the presence of C.trachomatis by polymerase chain reaction (PCRand ELISA.Materials and Methods: Endocervical swabs were collected from 80 women; 22 of them wereasymptomatic and 58 symptomatic. Samples were examined by PCR designed to detect Chlamydialplasmid using specific KL1 and KL2 primers. Serum IgG and IgA antibodies to C.trachomatiswere detected by ELISA. Since elevated CRP levels are a marker for inflammation, the presence ofC- Reactive protein (CRP has also been evaluated in all samples.Results: The rate of C.trachomatis infection by PCR was revealed to be 27.2% and 18.9% inasymptomatic and symptomatic women, respectively The χ2 test shows no significant difference(p value= 0.22. Serological screening was done on all samples. The high level of IgG and IgAto C.trachomatis infection was 29.4% and 17.6%, respectively. The presence of high levels ofCRP, as a serological marker of persistence infection, was 31.8% and 34.4% in asymptomaticand symptomatic women, respectively. The high rate of CRP level in the samples indicates acuteinfections in both groups.Conclusion: Genital C.trachomatis infection is the leading cause of tubal factor infertility. Thepresent study shows that C.trachomatis infection could be present in symptomatic as well asasymptomatic women. Therefore, a screening test for C.trachomatis infection is recommended forall women who refer to gynecologic outpatient departments in Isfahan and other parts of Iran.

  13. The effects of selected drugs, including chlorpromazine and non-steroidal anti-inflammatory agents, on polyclonal IgG synthesis and interleukin 1 production by human peripheral blood mononuclear cells in vitro.

    Science.gov (United States)

    Martinez, F; Coleman, J W

    1989-01-01

    We tested a range of drugs for their effects on in vitro polyclonal IgG synthesis by human peripheral blood mononuclear cells (PBMC) stimulated with the lectin pokeweed mitogen (PWM). The test drugs were selected on the basis of reported disruptive effects on immune function in vivo. IgG production between day 4 and days 7 or 8 of culture was measured by biotin-streptavidin sandwich ELISA. The anti-psychotic agent chlorpromazine (0.55-1.7 microM) enhanced IgG synthesis to approximately double control levels. In contrast, the non-steroidal anti-inflammatory drugs (NSAIDs) indomethacin, piroxicam, ibuprofen and aspirin inhibited IgG synthesis by up to 50%, with a rank order of potency that reflects their activity as inhibitors of cyclo-oxygenase. Phenytoin, procainamide, propylthiouracil, methimazole, D-penicillamine and D-penicillamine-L-cysteine all failed to modulate IgG synthesis at non-toxic concentrations. The potentiation and inhibition of IgG synthesis by chlorpromazine and indomethacin, respectively, was observed only when the drug was present during the first 24 h of culture. Neither chlorpromazine nor indomethacin, at non-toxic concentrations, affected PHA- and PWM-stimulated proliferation of PBMC. In addition, chlorpromazine, indomethacin and piroxicam, at concentrations which produced maximal modulation of IgG synthesis, and D-penicillamine and D-penicillamine-L-cysteine at 10 microM failed to influence production of interleukin-1-like activity. We conclude that chlorpromazine and NSAIDs, although they exert opposite effects on IgG synthesis, act at an early stage of B cell differentiation that appears to be independent of interleukin 1 synthesis and early proliferative events. PMID:2788047

  14. A dual-monoclonal sandwich ELISA specific for hepcidin-25.

    Science.gov (United States)

    Butterfield, Anthony M; Luan, Peng; Witcher, Derrick R; Manetta, Joseph; Murphy, Anthony T; Wroblewski, Victor J; Konrad, Robert J

    2010-11-01

    Hepcidin, a key regulator of iron metabolism, binds to the iron transporter ferroportin to cause its degradation. In humans, hepcidin deficiency has been linked to hemochromatosis and iron overload, whereas increased concentrations have been reported in anemia of cancer and chronic disease. There is currently an unmet clinical need for a specific immunoassay with a low limit of quantification to measure serum concentrations of hepcidin-25, the active form of the protein. We generated 2 antihepcidin-25 monoclonal antibodies and used them to build a sandwich ELISA. We correlated ELISA results to hepcidin-25 measurements by LC-MS and used ELISA to measure serum hepcidin-25 concentrations in normal individuals, cancer patients, and patients with rheumatoid arthritis. The sandwich ELISA was highly specific for hepcidin-25, having a limit of quantification of 0.01 μg/L (10 pg/mL). Serum concentrations of hepcidin-25 measured by ELISA correlated with hepcidin-25 concentrations measured by using an independent LC-MS assay (r = 0.98, P < 0.001). Hepcidin-25 concentrations were increased in patients with cancer (median 54.8 μg/L, 25%-75% range 23.2-93.5 μg/L, n = 34) and rheumatoid arthritis (median 10.6 μg/L, 25%-75% range 5.9-18.4 μg/L, n = 76) compared with healthy individuals (median 1.20 μg/L, 25%-75% range 0.42-3.07 μg/L, n = 100). The use of 2 monoclonal antibodies in a sandwich ELISA format provides a robust and convenient method for measuring concentrations of the active form of hepcidin. This ELISA should help to improve our understanding of the role of hepcidin in regulating iron metabolism.

  15. Immunoglobulin G Avidities in Infants in Mexico after Primary Immunization with Three Doses of Polyribosylribitol Phosphate-Tetanus Toxoid Haemophilus influenzae Type b Vaccine▿ †

    Science.gov (United States)

    Gómez-de-León, Patricia; Díaz-García, F. Javier; Villaseñor-Sierra, Alberto; Segura, Jorge; Carranza, Martha I.; Arredondo-Garcia, José Luis; Santos, José Ignacio

    2008-01-01

    Serum immunoglobulin G concentrations and avidities specific to Haemophilus influenzae type b (Hib) were measured in 208 children living in Guadalajara and Mexico City. Protective concentrations were found in 98.9% and 100.0% of participants, respectively. Geometric mean concentrations differed between both populations and/or among age groups. Mean avidities differed only among the 7- to 12-month-old children. Diphtheria-tetanus-whole-cell pertussis-hepatitis B-Hib primary vaccination seems to induce protection in Mexican children. PMID:18417667

  16. Immunoglobulin G avidities in infants in Mexico after primary immunization with three doses of polyribosylribitol phosphate-tetanus toxoid Haemophilus influenzae type b vaccine.

    Science.gov (United States)

    Gómez-de-León, Patricia; Díaz-García, F Javier; Villaseñor-Sierra, Alberto; Segura, Jorge; Carranza, Martha I; Arredondo-Garcia, José Luis; Santos, José Ignacio

    2008-06-01

    Serum immunoglobulin G concentrations and avidities specific to Haemophilus influenzae type b (Hib) were measured in 208 children living in Guadalajara and Mexico City. Protective concentrations were found in 98.9% and 100.0% of participants, respectively. Geometric mean concentrations differed between both populations and/or among age groups. Mean avidities differed only among the 7- to 12-month-old children. Diphtheria-tetanus-whole-cell pertussis-hepatitis B-Hib primary vaccination seems to induce protection in Mexican children.

  17. Comparison between a Conductometric Biosensor and ELISA in the Evaluation of Johne’s Disease

    Directory of Open Access Journals (Sweden)

    Chika Okafor

    2014-10-01

    Full Text Available Johne’s disease (JD, caused by Mycobacterium avium subspecies paratuberculosis (MAP, is an important gastrointestinal disease of cattle worldwide because of the economic losses encountered in JD-affected herds. These losses include reduction in milk yield in cows, premature culling and reduced carcass weight of culled diseased animals. In the U.S. dairy industry, economic losses from reduced productivity associated with JD are estimated to cost between $200 and $250 million annually. The development of non-laboratory-based assays would support more frequent testing of animals for JD and could improve its control. Conductometric biosensors combine immunomigration technology with electronic signal detection and have been adapted for the detection of IgG antibody against MAP. In the present study, a capture membrane with limited variability in the immunomigration channel and an optimal concentration of the secondary anti-bovine antibody used in a previously developed conductometric biosensor were compared with a commercially available antibody detection ELISA in their evaluation of JD, using samples of serum from cattle whose JD status where unknown. There was a moderate strength of agreement (kappa = 0.41 between the two assays. Findings from this preliminary study support the continued development of conductometric biosensors for use in the diagnosis of JD.

  18. IgG4-unrelated type 1 autoimmune pancreatitis

    Science.gov (United States)

    Nakano, Eriko; Kanno, Atsushi; Masamune, Atsushi; Yoshida, Naoki; Hongo, Seiji; Miura, Shin; Takikawa, Tetsuya; Hamada, Shin; Kume, Kiyoshi; Kikuta, Kazuhiro; Hirota, Morihisa; Nakayama, Keisuke; Fujishima, Fumiyoshi; Shimosegawa, Tooru

    2015-01-01

    A 50-year-old male was referred to our hospital for the evaluation of hyperproteinemia. Fluorodeoxyglucose positron emission tomography revealed high fluorodeoxyglucose uptake in the pancreas, bilateral lacrimal glands, submandibular glands, parotid glands, bilateral pulmonary hilar lymph nodes, and kidneys. Laboratory data showed an elevation of hepatobiliary enzymes, renal dysfunction, and remarkably high immunoglobulin (Ig) G levels, without elevated serum IgG4. Abdominal computed tomography revealed swelling of the pancreatic head and bilateral kidneys. Endoscopic retrograde cholangiopancreatography showed an irregular narrowing of the main pancreatic duct in the pancreatic head and stricture of the lower common bile duct. Histological examination by endoscopic ultrasonography-guided fine-needle aspiration revealed findings of lymphoplasmacytic sclerosing pancreatitis without IgG4-positive plasma cells. Abnormal laboratory values and the swelling of several organs were improved by the treatment with steroids. The patient was diagnosed as having type 1 autoimmune pancreatitis (AIP) based on the International Consensus Diagnostic Criteria. Therefore, we encountered a case of compatible type 1 AIP without elevated levels of serum IgG4 or IgG4-positive plasma cells. This case suggests that AIP phenotypes are not always associated with IgG4. PMID:26361429

  19. Structural characterization of the Man5 glycoform of human IgG3 Fc

    Energy Technology Data Exchange (ETDEWEB)

    Shah, Ishan S.; Lovell, Scott; Mehzabeen, Nurjahan; Battaile, Kevin P.; Tolbert, Thomas J. (Kansas); (HWMRI)

    2017-12-01

    Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8 Å resolution and compared its structural features with published structures from the other IgG subclasses. Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). The IgG3 Fc structure helps to explain some of these differences. Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function.

  20. Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres Indirect ELISA for the serological diagnosis of visceral leishmaniasis in wild canids

    Directory of Open Access Journals (Sweden)

    Paulo R.B. Ferreira

    2013-04-01

    assay (ELISA for the serodiagnosis of visceral leishmaniasis (VL in Brazilian species of wild canids is described. Serum and plasma samples from 12 captive wild canids were studied: seven from maned wolves (Chrysocyon brachyurus, three from hoary foxes (Lycalopex vetulus, and two from crab-eating foxes (Cerdocyon thous. Samples from C. brachyurus and L. vetulus, both captive in an endemic area for VL, presenting clinical disease and positivity in Indirect Immunofluorescence Reaction and Polymerase Chain Reaction tests were used as positive controls. The antibody anti-dog IgG and Protein A, both conjugated with horseradish peroxidase, were compared in indirect ELISA tests which detected four (04/12 and three (03/12 seropositive C. brachyurus for anti-Leishmania antibodies, respectively. The ELISA tests were able to clearly distinguish negative from positive samples, as the mean optical density (OD of the negative samples was 4.8 and 15.5 times lower than those of the positive ones either using anti-dog IgG and Protein A, respectively. Samples from three ELISA - positive C. brachyurus were analyzed by Western blotting and identified immunodominant bands of 19, 22, 24, 45 and 66 kDa, among 22 protein bands detected. The ELISAs with protein A and anti-dog IgG showed respectively excellent (Kappa = 1.0; p<0.001 and moderate (Kappa = 0.8; p<0.0015 agreement with the Western blotting assay. The ELISA tests showed to be adequate for screening studies to identify antibody responses, thus indicating contact with Leishmania infection by wild canids.

  1. Synergistic Inhibition of R5 HIV-1 by the Fusion Protein (FLSC) IgG1 Fc and Maraviroc in Primary Cells: Implications for Prevention and Treatment.

    Science.gov (United States)

    Latinovic, Olga S; Zhang, Jian; Tagaya, Yutaka; DeVico, Anthony L; Fouts, Timothy R; Schneider, K; Lakowicz, Joseph R; Heredia, Alonso; Redfield, Robert R

    2016-01-01

    Antiretroviral (ARV) drugs targeting retroviral enzymes have been extensively employed to treat HIV-1 infection. Drawbacks of this approach include cost, toxicity, and the eventual emergence of resistant strains that threaten prophylactic and/or therapeutic efficacy. Accordingly, efforts to develop next-generation ARV approaches are warranted, particularly if they can offer a higher threshold of resistance. We have previously shown that FLSC, a fusion protein containing gp120(BAL) and the D1 and D2 domains of human CD4, specifically binds CCR5, an important cellular co-receptor, and inhibits the entry of R5 HIV isolates. (FLSC) IgG1, a fusion of FLSC and the hinge-C(H)2-C(H)3 region of human IgG1, has an increased antiviral activity, likely due to the resultant bivalency. In this study, we show CCR5 reduction upon (FLSC) IgG1 treatment both by standard flow cytometry and visualized using a novel nanoparticle method. A β-lactamase virus-cell fusion assay was used to quantify (FLSC) IgG1 inhibition of HIV-1 entry into both cell lines and primary cells. Synergistic anti-viral activities of (FLSC) IgG1 and MVC in primary cells were evaluated by measuring supernatant p24 levels via ELISA and calculated using the MacSynergy™ II program. We previously reported that treatment with the CCR5 small molecule antagonist Maraviroc (MVC) increased the apparent exposure of the (FLSC) IgG1 binding sites on CCR5, leading us to wonder if the two compounds used in combination might synergize in their anti-viral activity. Here we show that this is indeed the case. We demonstrate that fusion protein (FLSC) IgG1, strongly synergizes with the CCR5 antagonist Maraviroc to successfully inhibit both MVC-sensitive and MVC-resistant R5 HIV-1. Observed synergy between (FLSC) IgG1 and MVC was high in both, cell lines and primary PBMCs. This has relevance for future in vivo studies. In addition, synergy occurred both with MVC-sensitive viruses and MVC-resistant viruses, partially restoring the

  2. Fifth Percentile Cutoff Values for Antipneumococcal Polysaccharide and Anti-Salmonella typhi Vi IgG Describe a Normal Polysaccharide Response

    Directory of Open Access Journals (Sweden)

    Heidi Schaballie

    2017-05-01

    Full Text Available BackgroundSerotype-specific antibody responses to unconjugated pneumococcal polysaccharide vaccine (PPV evaluated by a World Health Organization (WHO-standardized enzyme-linked immunosorbent assay (ELISA are the gold standard for diagnosis of specific polysaccharide antibody deficiency (SAD. The American Academy of Allergy, Asthma and Immunology (AAAAI has proposed guidelines to interpret the PPV response measured by ELISA, but these are based on limited evidence. Additionally, ELISA is costly and labor-intensive. Measurement of antibody response to Salmonella typhi (S. typhi Vi vaccine and serum allohemagglutinins (AHA have been suggested as alternatives. However, there are no large cohort studies and cutoff values are lacking.ObjectiveTo establish cutoff values for antipneumococcal polysaccharide antibody response, anti-S. typhi Vi antibody, and AHA.MethodsOne hundred healthy subjects (10–55 years were vaccinated with PPV and S. typhi Vi vaccine. Blood samples were obtained prior to and 3–4 weeks after vaccination. Polysaccharide responses to 3 serotypes were measured by WHO ELISA and to 12 serotypes by an in-house bead-based multiplex assay. Anti-S. typhi Vi IgG were measured with a commercial ELISA kit. AHA were measured by agglutination method.ResultsApplying AAAAI criteria, 30% of healthy subjects had a SAD. Using serotype-specific fifth percentile (p5 cutoff values for postvaccination IgG and fold increase pre- over postvaccination, only 4% of subjects had SAD. One-sided 95% prediction intervals for anti-S. typhi Vi postvaccination IgG (≥11.2 U/ml and fold increase (≥2 were established. Eight percent had a response to S. typhi Vi vaccine below these cutoffs. AHA titer p5 cutoffs were ½ for anti-B and ¼ for anti-A.ConclusionWe establish reference cutoff values for interpretation of PPV response measured by bead-based assay, cutoff values for S. typhi Vi vaccine responses, and normal values for AHA. For the first time, the

  3. ELISA test for p63 antibodies in chronic ulcerative stomatitis.

    Science.gov (United States)

    Solomon, L W; Stark, P C; Winter, L; Kumar, V; Sinha, S

    2010-03-01

    To develop a novel test for chronic ulcerative stomatitis (CUS), a chronic immunologically mediated condition that produces oral ulcerations. Current diagnostic methods require expensive and technically demanding in situ immunofluorescence (IF) studies. An Enzyme-Linked ImmunoSorbent Assay (ELISA) was prepared and tested with serum samples from patients with CUS and negative controls. The N-terminal portion of the CUS autoantigen, DeltaNp63alpha, was produced as a purified recombinant protein and used to coat ELISA plates. Sera from 25 patients with CUS and 16 negative controls were analyzed for reactive antibodies. The optimal cut-offs for positive and negative samples were determined. The optimal cut-off of 0.236 resulted in a sensitivity and specificity of the ELISA of 0.80 and 0.75, respectively (exact 95% confidence intervals, P-value of <0.001). The ELISA developed in this study provides a novel and reliable diagnostic assessment to distinguish CUS from other oral ulcerative diseases. Immunoassay will allow the true incidence and prevalence of CUS to be determined in future studies. When combined with clinical correlations, the ELISA results will facilitate the evaluation of the prognostic utility of antibody titers and allow correlation with treatment responses in individual CUS cases.

  4. Hybrid IgG4/IgG4 Fc antibodies form upon 'Fab-arm' exchange as demonstrated by SDS-PAGE or size-exclusion chromatography

    NARCIS (Netherlands)

    Rispens, Theo; den Bleker, Tamara H.; Aalberse, Rob C.

    2010-01-01

    Human IgG4 antibodies are dynamic molecules that in vivo exchange half-molecules to become bispecific antibodies. Here we show that IgG4 antibodies and IgG4 Fc fragments similarly exchange resulting in hybrid antibodies (a single Fab + Fc) with a molecular weight of ca. 100 kDa. These antibodies can

  5. Pachymeningitis associated with IgG4 disease

    Directory of Open Access Journals (Sweden)

    Dan Zimelewicz Oberman

    2017-06-01

    Full Text Available Hypertrophic pachymeningitis is an infrequent disorder. It can be idiopathic or secondary to infectious, autoimmune or neoplastic disease. The recently described 〈IgG4-related disease〉 could be the origin of many cases considered cryptogenic. We present the case of a 60-year-old man, with a history of headache and episcleritis in both eyes, with partial response to corticoid therapy. The brain MR study with gadolinium showed enhancement and thickening of the dura mater, extending from lateral wall of left temporal and occipital lobes to ipsilateral tentorium. Meningeal biopsy showed fibrosis and lymphoplasmacytic infiltrate, with more than 10 IgG4+ plasma cells per high power field. After treatment with rituximab there was clinical improvement accompanied by the virtual disappearance of the alterations detected in neuroimaging. Hypertrophic pachymeningitis as a manifestation of IgG4-related disease can be based on MRI findings if plasma IgG4 are elevated.

  6. Entamoeba histolytica: detection of coproantigens by purified antibody in the capture sandwich ELISA Entamoeba histolytica: detecção de coproantígenos por ELISA de captura utilizando anticorpo purificado

    Directory of Open Access Journals (Sweden)

    Haidee Urdaneta

    1994-12-01

    Full Text Available A sensitive and specific Capture Sandwich ELISA (CSE was developed using polyclonal purified rabbit antibodies against three different axenic strains of Entamoeba histolytica: CSP from Brazil and HM1 - IMSS from Mexico, for the detection of coproantigens in fecal samples. Immunoglobulin G (IgG againstis E. histolytica was isolated from rabbits immunized with throphozoites whole extract in two stages: affinity chromatography in a column containing E. histolytica antigens bound to Sepharose 4B was followed by another chromatography in Sepharose antibodies 4B-Protein A. A Capture Sandwich ELISA using purified antibodies was able to detect 70ng of amebae protein, showing a sensitivity of 93% and specificity of 94%. The combination of microscopic examination and CSE gave a concordance and discordance of 93.25% and 6.75%, respectively. It was concluded that CSE is highly specific for the detection of coproantigens of E. histolytica in feces of infected patients, is quicker to perform, easier and more sensitive than microscopic examination.Foi desenvolvido um teste de ELISA de Captura usando anticorpos policlonais purificados obtidos em coelhos contra três diferentes cepas axênicas de Entamoeba histolytica (ICB-CSP and ICB-462 do Brasil e HM1 do México para detecção de coproantígenos em amostras de fezes de indivíduos: a sintomáticos, b assintomáticos, c com outros parasitos intestinais, e d sadios. Imunoglobulina G (IgG contra E. histolytica foi isolada de imune soro de coelho, em duas etapas: cromatografia de afinidade em uma coluna contendo antígenos de E. histolytica unidos à Sepharose 4B, seguido por outra cromatografía em Sepharose 4B Proteína A. O teste de ELISA usando anticorpos purificados, foi capaz de detectar até um só trofozoíto por lâmina ou 70 ng de proteína de ameba por orifício, apresentando uma sensibilidade de 93% e uma especificidade de 94%. A combinação do exame microscópico com o método de ELISA de Captura

  7. Immunohistochemical Characteristics of IgG4-Related Tubulointerstitial Nephritis: Detailed Analysis of 20 Japanese Cases

    Directory of Open Access Journals (Sweden)

    Mitsuhiro Kawano

    2012-01-01

    Full Text Available Although tubulointerstitial nephritis with IgG4+ plasma cell (PC infiltration is a hallmark of IgG4-related kidney disease (IgG4-RKD, only a few studies are available about the minimum number of IgG4+ PC needed for diagnosis along with IgG4+/IgG+ PC ratio in the kidney. In addition, the significance of the deposition of IgG or complement as a reflection of humoral immunity involvement is still uncertain. In this study, we analyzed 20 Japanese patients with IgG4-RKD to evaluate the number of IgG4+ PCs along with IgG4+/IgG+ PC ratio and involvement of humoral immunity. The average number of IgG4+ PCs was 43.8/hpf and the average IgG4+/IgG+ or IgG4+/CD138+ ratio was 53%. IgG and C3 granular deposits on the tubular basement membrane (TBM were detected by immunofluorescence microscopy in 13% and 47% of patients, respectively. Nine patients had a variety of glomerular lesions, and 7 of them had immunoglobulin or complement deposition in the glomerulus. In conclusion, we confirmed that infiltrating IgG4+ PCs > 10/hpf and/or IgG4/IgG (CD138+ PCs > 40% was appropriate as an item of the diagnostic criteria for IgG4-RKD. A relatively high frequency of diverse glomerular lesions with immunoglobulin or complement deposits and deposits in TBM may be evidence of immune complex involvement in IgG4-related disease.

  8. Recalibration of the limiting antigen avidity EIA to determine mean duration of recent infection in divergent HIV-1 subtypes.

    Directory of Open Access Journals (Sweden)

    Yen T Duong

    Full Text Available Mean duration of recent infection (MDRI and misclassification of long-term HIV-1 infections, as proportion false recent (PFR, are critical parameters for laboratory-based assays for estimating HIV-1 incidence. Recent review of the data by us and others indicated that MDRI of LAg-Avidity EIA estimated previously required recalibration. We present here results of recalibration efforts using >250 seroconversion panels and multiple statistical methods to ensure accuracy and consensus.A total of 2737 longitudinal specimens collected from 259 seroconverting individuals infected with diverse HIV-1 subtypes were tested with the LAg-Avidity EIA as previously described. Data were analyzed for determination of MDRI at ODn cutoffs of 1.0 to 2.0 using 7 statistical approaches and sub-analyzed by HIV-1 subtypes. In addition, 3740 specimens from individuals with infection >1 year, including 488 from patients with AIDS, were tested for PFR at varying cutoffs.Using different statistical methods, MDRI values ranged from 88-94 days at cutoff ODn = 1.0 to 177-183 days at ODn = 2.0. The MDRI values were similar by different methods suggesting coherence of different approaches. Testing for misclassification among long-term infections indicated that overall PFRs were 0.6% to 2.5% at increasing cutoffs of 1.0 to 2.0, respectively. Balancing the need for a longer MDRI and smaller PFR (<2.0% suggests that a cutoff ODn = 1.5, corresponding to an MDRI of 130 days should be used for cross-sectional application. The MDRI varied among subtypes from 109 days (subtype A&D to 152 days (subtype C.Based on the new data and revised analysis, we recommend an ODn cutoff = 1.5 to classify recent and long-term infections, corresponding to an MDRI of 130 days (118-142. Determination of revised parameters for estimation of HIV-1 incidence should facilitate application of the LAg-Avidity EIA for worldwide use.

  9. Recalibration of the limiting antigen avidity EIA to determine mean duration of recent infection in divergent HIV-1 subtypes.

    Science.gov (United States)

    Duong, Yen T; Kassanjee, Reshma; Welte, Alex; Morgan, Meade; De, Anindya; Dobbs, Trudy; Rottinghaus, Erin; Nkengasong, John; Curlin, Marcel E; Kittinunvorakoon, Chonticha; Raengsakulrach, Boonyos; Martin, Michael; Choopanya, Kachit; Vanichseni, Suphak; Jiang, Yan; Qiu, Maofeng; Yu, Haiying; Hao, Yan; Shah, Neha; Le, Linh-Vi; Kim, Andrea A; Nguyen, Tuan Anh; Ampofo, William; Parekh, Bharat S

    2015-01-01

    Mean duration of recent infection (MDRI) and misclassification of long-term HIV-1 infections, as proportion false recent (PFR), are critical parameters for laboratory-based assays for estimating HIV-1 incidence. Recent review of the data by us and others indicated that MDRI of LAg-Avidity EIA estimated previously required recalibration. We present here results of recalibration efforts using >250 seroconversion panels and multiple statistical methods to ensure accuracy and consensus. A total of 2737 longitudinal specimens collected from 259 seroconverting individuals infected with diverse HIV-1 subtypes were tested with the LAg-Avidity EIA as previously described. Data were analyzed for determination of MDRI at ODn cutoffs of 1.0 to 2.0 using 7 statistical approaches and sub-analyzed by HIV-1 subtypes. In addition, 3740 specimens from individuals with infection >1 year, including 488 from patients with AIDS, were tested for PFR at varying cutoffs. Using different statistical methods, MDRI values ranged from 88-94 days at cutoff ODn = 1.0 to 177-183 days at ODn = 2.0. The MDRI values were similar by different methods suggesting coherence of different approaches. Testing for misclassification among long-term infections indicated that overall PFRs were 0.6% to 2.5% at increasing cutoffs of 1.0 to 2.0, respectively. Balancing the need for a longer MDRI and smaller PFR (<2.0%) suggests that a cutoff ODn = 1.5, corresponding to an MDRI of 130 days should be used for cross-sectional application. The MDRI varied among subtypes from 109 days (subtype A&D) to 152 days (subtype C). Based on the new data and revised analysis, we recommend an ODn cutoff = 1.5 to classify recent and long-term infections, corresponding to an MDRI of 130 days (118-142). Determination of revised parameters for estimation of HIV-1 incidence should facilitate application of the LAg-Avidity EIA for worldwide use.

  10. A Peptide Mimetic of 5-Acetylneuraminic Acid-Galactose Binds with High Avidity to Siglecs and NKG2D.

    Directory of Open Access Journals (Sweden)

    Laura L Eggink

    Full Text Available We previously identified several peptide sequences that mimicked the terminal sugars of complex glycans. Using plant lectins as analogs of lectin-type cell-surface receptors, a tetravalent form of a peptide with the sequence NPSHPLSG, designated svH1C, bound with high avidity to lectins specific for glycans with terminal 5-acetylneuraminic acid (Neu5Ac-galactose (Gal/N-acetylgalactosamine (GalNAc sequences. In this report, we show by circular dichroism and NMR spectra that svH1C lacks an ordered structure and thus interacts with binding sites from a flexible conformation. The peptide binds with high avidity to several recombinant human siglec receptors that bind preferentially to Neu5Ac(α2,3Gal, Neu5Ac(α2,6GalNAc or Neu5Ac(α2,8Neu5Ac ligands. In addition, the peptide bound the receptor NKG2D, which contains a lectin-like domain that binds Neu5Ac(α2,3Gal. The peptide bound to these receptors with a KD in the range of 0.6 to 1 μM. Binding to these receptors was inhibited by the glycoprotein fetuin, which contains multiple glycans that terminate in Neu5Ac(α2,3Gal or Neu5Ac(α2,6Gal, and by sialyllactose. Binding of svH1C was not detected with CLEC9a, CLEC10a or DC-SIGN, which are lectin-type receptors specific for other sugars. Incubation of neuraminidase-treated human peripheral blood mononuclear cells with svH1C resulted in binding of the peptide to a subset of the CD14+ monocyte population. Tyrosine phosphorylation of siglecs decreased dramatically when peripheral blood mononuclear cells were treated with 100 nM svH1C. Subcutaneous, alternate-day injections of svH1C into mice induced several-fold increases in populations of several types of immune cells in the peritoneal cavity. These results support the conclusion that svH1C mimics Neu5Ac-containing sequences and interacts with cell-surface receptors with avidities sufficient to induce biological responses at low concentrations. The attenuation of inhibitory receptors suggests that svH1C

  11. ELISA reader does not interfere by mobile phone radiofrequency radiation

    Science.gov (United States)

    Mortazavi, Seyyed Mohammad Javad; Baradaran-Ghahfarokhi, Hamid Reza; Abdi, Mohammad Reza; Baradaran-Ghahfarokhi, Milad; Mostafavi, Nayyer Sadat; Mahmoudi, Golshan; Berenjkoub, Nafiseh; Akmali, Zahra; Hossein-Beigi, Fahimeh; Arsang, Vajiheh

    2016-01-01

    Background: The increasing number of mobile phones can physically cause electromagnetic interference (EMI) in medical environments; can also cause errors in immunoassays in laboratories. The ELISA readers are widely used as a useful diagnostic tool for Enzymun colorimetric assay in medicine. The aim of this study was to investigate whether the ELISA reader could be interfered by the exposure to the 900 MHz cell phones in the laboratory. Materials and Methods: Human serum samples were collected from 14 healthy donors (9 women and 5 men) and each sample was divided into four aliquots and was placed into four batches for the in-vitro quantitative determination of human chorionic gonadotropin (hCG). During colorimetric reading of the first, second, and third batches, the ELISA reader (Stat Fax 2100, Awareness Technology, Inc., USA) was exposed to 0.5, 1.0, and 2.0 W exposure of 900 MHz radiation, respectively. For the forth batch (control group), no radiation was applied. All experiments were performed comparing ELISA read out results of the I, II, and III batches with the control batch, using the Wilcoxon test with criterion level of P = 0.050. Results: The final scores in the exposed batches I, II, and III were not statistically significant relative to the control batch (P > 0.05). The results showed that 900 MHz radiation exposure did not alter the ELISA measured levels of hCG hormone in I (P = 0.219), II (P = 0.909), and III (P = 0.056) batches compared to the control batch. Conclusion: This study showed that ELISA reader does not interfere by mobile phone RF radiation at a closed contact (less than 5 cm distance). However, we recommend that medical institutions discuss these issues in the context of their specific use of technologies and frame a policy that is clear and straightforward to guide staff, patients, and visitors. PMID:27376040

  12. Evaluation of SD BIOLINEH. pyloriAg rapid test against double ELISA with SDH. pyloriAg ELISA and EZ-STEPH. pyloriAg ELISA tests.

    Science.gov (United States)

    Negash, Markos; Kassu, Afework; Amare, Bemnet; Yismaw, Gizachew; Moges, Beyene

    2018-01-01

    Helicobacter pylori antibody titters fall very slowly even after successful treatment. Therefore, tests detecting H. pylori antibody lack specificity and sensitivity. On the other hand, H. pylori stool antigen tests are reported as an alternative assay because of their reliability and simplicity. However, the comparative performance of H. pylori stool antigen tests for detecting the presence of the bacterium in clinical specimens in the study area is not assessed. Therefore, in this study we evaluated the performance of SD BIOLINE H. pylori Ag rapid test with reference to the commercially available EZ- STEP ELISA and SD BIOLINE H. pylori Ag ELISA tests. Stool samples were collected to analyse the diagnostic performance of SD BIOLINE H. pylori Ag rapid test kit using SD H. pylori Ag ELISA kit and EZ- STEP ELISA tests as a gold standard. Serum samples were also collected from each patient to test for the presence of H. pylori antibodies using dBest H. pylori Test Disk. Sensitivity, specificity, predictive values and kappa value are assessed. P values rapid test were: 95.6% (95% CI, 88.8-98.8), 92.5% (95%CI, 89-94.1%), 86.7% (95% CI, 80.5-89.6), and 97.6% (95% CI, 993.9-99.3) respectively. The performance of SD BIOLINE H. pylori Ag rapid test was better than the currently available antibody test in study area. Therefore, the SD BIOLINE Ag rapid stool test could replace and be used to diagnose active H. pylori infection before the commencement of therapy among dyspeptic patients.

  13. An ELISA for the quantitation of von Willebrand Factor

    DEFF Research Database (Denmark)

    Vinholt, Pernille Just; Overgaard, Martin; Diederichsen, Axel Cosmus Pyndt

    2013-01-01

    for measurement of von Willebrand factor-osteoprotegerin complex (VWF:OPG) in human plasma. Furthermore, the significance of VWF:OPG complex as a marker of cardiovascular disease (CVD) was evaluated. PATIENTS/METHODS: A sandwich ELISA for quantification of VWF:OPG was developed using a polyclonal rabbit anti...... with and without documented coronary calcification (total n=118). RESULTS AND CONCLUSIONS: The assay detected VWF:OPG complexes in human plasma, while no significant signal was observed when testing solutions containing VWF or recombinant OPG alone. Importantly, the ELISA assay was able to detect in vitro formed...

  14. Retratos falados em Elisa Lispector: um álbum fragmentado

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    André de Souza Pinto

    2015-11-01

    Full Text Available Este artigo, ao selecionar Retratos antigos: esboços a serem ampliados, de Elisa Lispector, teve como objetivo analisar a existência de um discurso fragmentado e memorialístico, assim como a construção, por intermédio dos retratos familiares, de uma autobiografia de Elisa e de sua família. Assim, o principal objetivo desse artigo foi apresentar a história dos Lispector feita por fotografias esquecidas em um velho álbum.

  15. IgE, IgG4 and IgA specific to Bet v 1-related food allergens do not predict oral allergy syndrome.

    Science.gov (United States)

    Guhsl, E E; Hofstetter, G; Lengger, N; Hemmer, W; Ebner, C; Fröschl, R; Bublin, M; Lupinek, C; Breiteneder, H; Radauer, C

    2015-01-01

    Birch pollen-associated plant food allergy is caused by Bet v 1-specific IgE, but presence of cross-reactive IgE to related allergens does not predict food allergy. The role of other immunoglobulin isotypes in the birch pollen-plant food syndrome has not been investigated in detail. Bet v 1-sensitized birch pollen-allergic patients (n = 35) were diagnosed for food allergy by standardized interviews, skin prick tests, prick-to-prick tests and ImmunoCAP. Concentrations of allergen-specific IgE, IgG1, IgG4 and IgA to seven Bet v 1-related food allergens were determined by ELISA. Bet v 1, Cor a 1, Mal d 1 and Pru p 1 bound IgE from all and IgG4 and IgA from the majority of sera. Immunoglobulins to Gly m 4, Vig r 1 and Api g 1.01 were detected in allergy and increased or reduced levels of IgE, IgG1, IgG4 or IgA specific to most Bet v 1-related allergens. Api g 1-specific IgE was significantly (P = 0.01) elevated in celeriac-allergic compared with celeriac-tolerant patients. Likewise, frequencies of IgE (71% vs 15%; P = 0.01) and IgA (86% vs 38%; P = 0.04) binding to Api g 1.01 were increased. Measurements of allergen-specific immunoglobulins are not suitable for diagnosing Bet v 1-mediated plant food allergy to hazelnut and Rosaceae fruits. In contrast, IgE and IgA to the distantly related allergen Api g 1 correlate with allergy to celeriac. © 2014 The Authors. Allergy Published by John Wiley & Sons Ltd.

  16. Purification of polyclonal IgG specific for Camelid’s antibodies and their recombinant nanobodies

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    Haddad Muhammad

    2016-01-01

    Full Text Available Camelid’ s heavy-chain antibody (HCAb consists of only two heavy chains and lacks the two light chains together with the CH1 domain usually found in conventional immunoglobulins. A recombinant single antigen-binding entity, named VHH (or Nanobody® was generated by reengineering the variable domains from HCAb. This study focuses on the detection of camelid´s immunoglobulins as well as their derivative nanobodies using a universal anti-camel antibody produced in rabbit (rIgG. Starting from a crude rabbit serum, a standard stock of rIgG (1 mg/ml was prepared after purification by affinity chromatography using protein-A column. As expected, rIgG was able to detect camel antibodies in ELISA and immunoblotting, and its reactivity was equal against all different camel IgG subclasses, which were purified from serum by differential affinity chromatography on protein-G and -A. Interestingly, rIgG also recognized nanobodies since they were originally part of camel HCAbs, providing an alternative method to detect the corpus of these recombinant proteins rather than targeting their artificial tags. These data suggest that the anti-camel rIgG described here could be efficiently applied at different stages of nanobody technology, including the quantitation of the issued nanobodies and their detection when bound to target antigens.

  17. Adult reference intervals for IgG subclasses with Siemens immunonephelometric assays in Chinese population.

    Science.gov (United States)

    Li, Ping; Liu, Zhongjuan; Wu, Ziyan; Wen, Xiaoting; Li, Liubing; Zhang, Shulan; Xu, Yingchun; Li, Yongzhe

    2017-01-01

    To determine the adult reference intervals for the Siemens IgG subclass reagents. 636 blood samples of healthy adults were analyzed to determine the level of IgG subclass using the reagents of Siemens immunonephelometric assay with molecular biology kits. IgGSc reference intervals were as follows: IgG1 4.45-9.76 g/L, IgG2 2.07-8.57 g/L, IgG3 0.08-0.80 g/L and IgG4 0.05-1.54 g/L. There was an excellent correlation between the total IgG and the sum of the IgG subclasses. No significant gender and age differences were observed. Our data provide the missing reference intervals and enable the use of the nephelometric IgG subclass reagents in Chinese. The study can offer reference on clinic diagnose.

  18. IgG2 immunodeficiency: association to pediatric patients with bacterial meningoencephalitis

    Directory of Open Access Journals (Sweden)

    ESCOBAR-PÉREZ XIOMARA

    2000-01-01

    Full Text Available An IgG subclass deficiency is often associated with bacterial infections. We studied four pediatric patients suffering from meningoencephalitis, two of them due to Streptococcus pneumoniae and two due to Haemophilus influenzae type b. Simultaneous diagnostic serum and cerebrospinal fluid samples were taken during income. The four subclasses of IgG and albumin were quantified in both biologic fluids by radial immunodiffusion. Very low levels of seric IgG2 with non detectable cerebrospinal fluid IgG2 were found in the patients. No intrathecal IgG subclass synthesis was found in two patients. One patient with S. pneumoniae had IgG3 intrathecal synthesis. Intrathecal IgG1, IgG3 and IgG4 synthesis was found in one patient suffering from H. influenzae according with reibergrams. Substitutive therapy with intravenous gammaglobulin was given to the patients as part of the treatment.

  19. What is IgG4? A review of the biology of a unique immunoglobulin subtype.

    Science.gov (United States)

    Nirula, Ajay; Glaser, Scott M; Kalled, Susan L; Taylor, Frederick R; Taylora, Frederick R

    2011-01-01

    Recent descriptions of the group of clinical disorders collectively defined as IgG4-related systemic disease (IgG4-RSD) have prompted this review of the unique biology of the IgG4 antibody. This article will discuss IgG4 structure and function, the unique phenomenon of half-antibody exchange, and the implications of IgG4 biology for its proposed role in immunologic diseases. IgG4 antibodies have unique structural and functional properties and undergo 'half-antibody exchange' in vivo, resulting in recombined antibodies composed of two different binding specificities. The production of IgG4 antibodies appears to be driven in part by T helper 2 (Th2) cytokines that mediate allergic responses and IgE production. Although serum IgG4 levels in healthy individuals vary significantly, data from multiple sclerosis (MS) patients suggest tight regulation of individual IgG4 levels over time. IgG4-RSD represents a diverse group of clinical disorders unified by elevated IgG4 levels and specific histopathologic findings. A key unanswered question is whether IgG4, a relatively weak activator of effector cells, is pathogenic in these disorders. IgG4 is a unique antibody biologically and structurally. Increased understanding of its precise role in the clinical syndromes that comprise IgG4-RSD may ultimately elucidate the underlying pathogenesis.

  20. Potential Mechanisms for IgG4 Inhibition of Immediate Hypersensitivity Reactions.

    Science.gov (United States)

    James, Louisa K; Till, Stephen J

    2016-03-01

    IgG4 is the least abundant IgG subclass in human serum, representing less than 5% of all IgG. Increases in IgG4 occur following chronic exposure to antigen and are generally associated with states of immune tolerance. In line with this, IgG4 is regarded as an anti-inflammatory antibody with a limited ability to elicit effective immune responses. Furthermore, IgG4 attenuates allergic responses by inhibiting the activity of IgE. The mechanism by which IgG4 inhibits IgE-mediated hypersensitivity has been investigated using a variety of model systems leading to two proposed mechanisms. First by sequestering antigen, IgG4 can function as a blocking antibody, preventing cross-linking of receptor bound IgE. Second IgG4 has been proposed to co-stimulate the inhibitory IgG receptor FcγRIIb, which can negatively regulate FcεRI signaling and in turn inhibit effector cell activation. Recent advances in our understanding of the structural features of human IgG4 have shed light on the unique functional and immunologic properties of IgG4. The aim of this review is to evaluate our current understanding of IgG4 biology and reassess the mechanisms by which IgG4 functions to inhibit IgE-mediated allergic responses.

  1. Historically aggressive types of follicular cell-derived thyroid cancer often have radioactive avid distant metastases: a study of 314 patients with distant metastases at a single institution

    Energy Technology Data Exchange (ETDEWEB)

    Tala, H.P.; Rondeau, G.; Fagin, J.A.; Tuttle, R.M. [Endocrinology Division, Department of Medicine, Nuclear Medicine Division, Memorial Sloan Kettering Cancer Center, New-York (United States); Ghossein, R.A. [Pathology Department, Nuclear Medecine Division, Memorial Sloan Kettering Cancer Center, New-York (United States); Grewal, R.K.; Larson, S.M. [Radiology Department, Nuclear Medicine Division, Memorial Sloan Kettering Cancer Center, New-York (United States)

    2012-07-01

    Radioactive iodine (RAI) remains one of the primary treatment options for metastatic, follicular cell derived thyroid cancers. The aim of this study was to determine the likelihood that metastatic lesions arising from one of the aggressive thyroid cancer histologies [tall cell variant of papillary thyroid carcinoma (TCV-PTC), poorly differentiated thyroid carcinoma (PDTC) and Hurthle cell carcinoma (HCC)] would demonstrate sufficient RAI avidity for visualization on RAI scanning and therefore could potentially benefit from RAI therapy. The study shows that in patients selected for RAI scanning or therapy at our center, RAI avid lesions can be identified in more than two thirds of the patients with distant metastases arising in the setting of C-PTC, WD-FTC, FV-PTC, TCV-PTC, or PDTC primary tumors. While RAI avidity on a post-therapy scan does not always correlate with clinically significant tumor killing activity, it is likely that some of these patients with RAI avid metastatic disease did obtain a clinical benefit

  2. Dual Testing Algorithm of BED-CEIA and AxSYM Avidity Index Assays Performs Best in Identifying Recent HIV Infection in a Sample of Rwandan Sex Workers

    NARCIS (Netherlands)

    Braunstein, S.L.; Nash, D.; Kim, A.A.; Ford, K.; Mwambarangwe, L.; Ingabire, C.M.; Vyankandondera, J.; van de Wijgert, J.H.H.M.

    2011-01-01

    To assess the performance of BED-CEIA (BED) and AxSYM Avidity Index (Ax-AI) assays in estimating HIV incidence among female sex workers (FSW) in Kigali, Rwanda. Eight hundred FSW of unknown HIV status were HIV tested; HIV-positive women had BED and Ax-AI testing at baseline and ≥12 months later to

  3. Colour measurement of colostrum for estimation of colostral IgG and colostrum composition in dairy cows.

    Science.gov (United States)

    Gross, Josef J; Kessler, Evelyne C; Bruckmaier, Rupert M

    2014-11-01

    Instruments for on-farm determination of colostrum quality such as refractometers and densimeters are increasingly used in dairy farms. The colour of colostrum is also supposed to reflect its quality. A paler or mature milk-like colour is associated with a lower colostrum value in terms of its general composition compared with a more yellowish and darker colour. The objective of this study was to investigate the relationships between colour measurement of colostrum using the CIELAB colour space (CIE L*=from white to black, a*=from red to green, b*=from yellow to blue, chroma value G=visual perceived colourfulness) and its composition. Dairy cow colostrum samples (n=117) obtained at 4·7±1·5 h after parturition were analysed for immunoglobulin G (IgG) by ELISA and for fat, protein and lactose by infrared spectroscopy. For colour measurements, a calibrated spectrophotometer was used. At a cut-off value of 50 mg IgG/ml, colour measurement had a sensitivity of 50·0%, a specificity of 49·5%, and a negative predictive value of 87·9%. Colostral IgG concentration was not correlated with the chroma value G, but with relative lightness L*. While milk fat content showed a relationship to the parameters L*, a*, b* and G from the colour measurement, milk protein content was not correlated with a*, but with L*, b*, and G. Lactose concentration in colostrum showed only a relationship with b* and G. In conclusion, parameters of the colour measurement showed clear relationships to colostral IgG, fat, protein and lactose concentration in dairy cows. Implementation of colour measuring devices in automatic milking systems and milking parlours might be a potential instrument to access colostrum quality as well as detecting abnormal milk.

  4. An indirect ELISA to detect the serologic response of elk (Cervus elaphus nelsoni) inoculated with Brucella abortus strain RB51.

    Science.gov (United States)

    Colby, Lesley A; Schurig, Gerhardt G; Elzer, Philip H

    2002-10-01

    An indirect enzyme-linked immunosorbent assay (ELISA) was developed to identify elk (Cervus elaphus nelsoni) with Brucella abortus strain RB51 (RB51)-specific antibodies using a mouse monoclonal antibody specific for bovine IgG1. This test was relatively easy to perform, accurate, and easily reproducible; therefore it could be standardized for use between laboratories. In addition, we attempted to compensate for inherent variabilities encountered when comparing ELISA readings from multiple samples taken from many animals over time. Optical density (OD) readings for each sample were converted into a percent positivity value for analysis. A negative cutoff value was determined above which a sample was considered to have a significantly elevated anti-RB51 antibody level. Pre- and postvaccination sera from 64 6-8 mo old elk, divided into four groups (females subcutaneously inoculated with saline (control animals), females ballistically inoculated with RB51, females subcutaneously inoculated with RB51, and males subcutaneously inoculated with RB51) were used. All serum samples were collected between 27 April and 15 November 1995. Values for all saline controls were appropriately below the negative cutoff value. All subcutaneously and ballistically inoculated elk were serologically positive to RB51 for at least two sampling periods during the study. The difference in percent positivity values for the ballistically compared to the subcutaneously inoculated groups was not statistically significant at 8, 10, 14, or 18 wk postvaccination. This suggests that processing RB51 into lactose based pellets and ballistically inoculating elk with these pellets does not alter the detectable elk antibody response. Also, inoculated and control animals can be accurately identified with ELISA at 4-8 weeks postvaccination.

  5. Quantitative modeling assesses the contribution of bond strengthening, rebinding and force sharing to the avidity of biomolecule interactions.

    Directory of Open Access Journals (Sweden)

    Valentina Lo Schiavo

    Full Text Available Cell adhesion is mediated by numerous membrane receptors. It is desirable to derive the outcome of a cell-surface encounter from the molecular properties of interacting receptors and ligands. However, conventional parameters such as affinity or kinetic constants are often insufficient to account for receptor efficiency. Avidity is a qualitative concept frequently used to describe biomolecule interactions: this includes incompletely defined properties such as the capacity to form multivalent attachments. The aim of this study is to produce a working description of monovalent attachments formed by a model system, then to measure and interpret the behavior of divalent attachments under force. We investigated attachments between antibody-coated microspheres and surfaces coated with sparse monomeric or dimeric ligands. When bonds were subjected to a pulling force, they exhibited both a force-dependent dissociation consistent with Bell's empirical formula and a force- and time-dependent strengthening well described by a single parameter. Divalent attachments were stronger and less dependent on forces than monovalent ones. The proportion of divalent attachments resisting a force of 30 piconewtons for at least 5 s was 3.7 fold higher than that of monovalent attachments. Quantitative modeling showed that this required rebinding, i.e. additional bond formation between surfaces linked by divalent receptors forming only one bond. Further, experimental data were compatible with but did not require stress sharing between bonds within divalent attachments. Thus many ligand-receptor interactions do not behave as single-step reactions in the millisecond to second timescale. Rather, they exhibit progressive stabilization. This explains the high efficiency of multimerized or clustered receptors even when bonds are only subjected to moderate forces. Our approach provides a quantitative way of relating binding avidity to measurable parameters including bond

  6. HIV Incidence Estimates Using the Limiting Antigen Avidity EIA Assay at Testing Sites in Kiev City, Ukraine: 2013-2014

    Science.gov (United States)

    Kruglov, Yuri; Yurchenko, Alexander

    2016-01-01

    Objective To estimate HIV incidence and highlight the characteristics of persons at greatest risk of HIV in the Ukraine capital, Kiev. Method Residual samples from newly-diagnosed persons attending the Kiev City AIDS Centre were tested for evidence of recent HIV infection using an avidity assay. Questions on possible risk factors for HIV acquisition and testing history were introduced. All persons (≥16yrs) presenting for an HIV test April’13–March’14 were included. Rates per 100,000 population were calculated using region-specific denominators. Results During the study period 6370 individuals tested for HIV. Of the 467 individuals newly-diagnosed with HIV, 21 had insufficient samples for LAg testing. Of the remaining 446, 39 (8.7%) were classified as recent with an avidity index <1.5ODn, 10 were reclassified as long-standing as their viral load was <1000 copies/mL, resulting in 29 (6.5%) recent HIV infections. The only independent predictor for a recent infection was probable route of exposure, with MSM more likely to present with a recent infection compared with heterosexual contact [Odds Ratio 8.86; 95%CI 2.65–29.60]. We estimated HIV incidence at 21.5 per 100,000 population, corresponding to 466 new infections. Using population estimates for MSM and PWID, incidence was estimated to be between 2289.6 and 6868.7/100,000 MSM, and 350.4 for PWID. Conclusion A high proportion of persons newly-infected remain undiagnosed, with MSM disproportionally affected with one in four newly-HIV-diagnosed and one in three recently-HIV-infected. Our findings should be used for targeted public health interventions and health promotion. PMID:27276170

  7. Seroprevalence of Rubella Specific IgG Antibody among Pregnant ...

    African Journals Online (AJOL)

    ABCCAFE7

    ABSTRACT: Rubella infection is generally mild in children. However, infection during pregnancy can leads to fetal death or congenital defects known as Congenital Rubella Syndrome. The seroprevalence of IgG to rubella virus was determined using Enzyme Linked Immunosorbent Assay among 400 pregnant women ...

  8. Seroprevalence of Rubella Specific IgG Antibody among Pregnant ...

    African Journals Online (AJOL)

    Rubella infection is generally mild in children. However, infection during pregnancy can leads to fetal death or congenital defects known as Congenital Rubella Syndrome. The seroprevalence of IgG to rubella virus was determined using Enzyme Linked Immunosorbent Assay among 400 pregnant women between the ages ...

  9. Seropositivity and determinants of immunoglobulin-G (IgG ...

    African Journals Online (AJOL)

    Objective: This study determined the seropositivity and determinants of serum IgG antibody against HSV-1 & HIV-2 among ... Conclusion: Our data shows that HSV-1 & 2 seropositivity among pregnant women in Port Harcourt is high; thus serolog- ical screening for ..... tation with certain risk and sociodemographic factors.

  10. ORIGINAL ARTICLE RUBELLA IgG ANTIBODY AMONG NIGERIAN ...

    African Journals Online (AJOL)

    boaz

    capable of causing a spectrum of birth defects described as congenital rubella syndrome (CRS). Despite the availability of safe and effective ... designed to determine the prevalence of anti- rubella IgG among pregnant women in order to ..... Vyse A.J., Gay N.J., White J.M., Ramsay. M.E., Brown D.W., Cohen B.J. Evolution.

  11. Increased IgG4 responses to multiple food and animal antigens indicate a polyclonal expansion and differentiation of pre-existing B cells in IgG4-related disease

    NARCIS (Netherlands)

    Culver, Emma L.; Vermeulen, Ellen; Makuch, Mateusz; van Leeuwen, Astrid; Sadler, Ross; Cargill, Tamsin; Klenerman, Paul; Aalberse, Rob C.; van Ham, S. Marieke; Barnes, Eleanor; Rispens, Theo

    2015-01-01

    IgG4-related disease (IgG4-RD) is a systemic fibroinflammatory condition, characterised by an elevated serum IgG4 concentration and abundant IgG4-positive plasma cells in the involved organs. An important question is whether the elevated IgG4 response is causal or a reflection of immune-regulatory

  12. Fragmentation of human IgG by a new protease isolated from the basidiomycete Armillaria mellea.

    Science.gov (United States)

    Hunneyball, I M; Stanworth, D R

    1975-01-01

    Digestion of human IgG by a new lysine-specific protease, isolated from the basidiomycete Armillaria mellea, produced Fc and Fab fragments similar to those produced by papain digestion of the same molecule. Digestion appeared to be restricted to a single cleavage point within the hinge region of the IgG molecule. Myeloma proteins of IgG1, IgG3 and IgG4 subclasses were found to be digested at an extremely rapid rate whereas IgG2 myeloma proteins appeared to be resistant to digestion by this enzyme. Images FIG. 2 FIG. 6 PMID:1201861

  13. A monoclonal blocking ELISA to detect chicken anaemia virus ...

    African Journals Online (AJOL)

    The blocking ELISA depends on the selective inhibition of the binding of MAb 2A9 to solid-phase antigen by CAV-specific antibodies present in convalescent chicken serum. Performance evaluation of the MBE using 417 sera from Nigerian and Northern Ireland commercial chicken flocks revealed a 99.3 % agreement ...

  14. Designing of enzyme linked immunosorbent assay (ELISA) kit for ...

    African Journals Online (AJOL)

    The sensitivity of microscopic examination of fecal samples to recognize Giardia parasites is low. In the methods based on antigen scanning of parasites such as enzyme linked immunosorbent assay (ELISA), copro-antigens of parasite will be traced and diagnosed even if the live parasite is absent in the fecal samples.

  15. Opportunities and challenges when pooling milk samples using ELISA

    DEFF Research Database (Denmark)

    Græsbøll, Kaare; Andresen, Lars Ole; Hisham Beshara Halasa, Tariq

    2017-01-01

    -positive samples by pooling. To illustrate this, the sensitivity of antibody ELISA on pooled samples of bovine milk for Salmonella Dublin, Mycobacterium avium spp. paratuberculosis, and bovine virus diarrhea was tested. For these milk assays, the analytical sensitivity decreased rapidly with increasing pool sizes...

  16. Detection of antibodies to the 20s proteasome by ELISA

    DEFF Research Database (Denmark)

    Jørgensen, Karin Meinike; Frederiksen, Jette Lautrup; Nielsen, Christoffer Tandrup

    2013-01-01

    The presence of antibodies against the 20S proteasome has been correlated with diseases like multiple sclerosis (MS) and systemic lupus erythematosus (SLE) but no definite association has been established. In order to investigate this further, we optimized an ELISA for proteasome antibodies...

  17. Evaluation of monoclonal antibody-based sandwich direct ELISA (MSD-ELISA for antigen detection of foot-and-mouth disease virus using clinical samples.

    Directory of Open Access Journals (Sweden)

    Kazuki Morioka

    Full Text Available A monoclonal antibody-based sandwich direct ELISA (MSD-ELISA method was previously developed for foot-and-mouth disease (FMD viral antigen detection. Here we evaluated the sensitivity and specificity of two FMD viral antigen detection MSD-ELISAs and compared them with conventional indirect sandwich (IS-ELISA. The MSD-ELISAs were able to detect the antigen in saliva samples of experimentally-infected pigs for a longer term compared to the IS-ELISA. We also used 178 RT-PCR-positive field samples from cattle and pigs affected by the 2010 type-O FMD outbreak in Japan, and we found that the sensitivities of both MSD-ELISAs were about 7 times higher than that of the IS-ELISA against each sample (P<0.01. In terms of the FMD-positive farm detection rate, the sensitivities of the MSD-ELISAs were about 6 times higher than that of the IS-ELISA against each farm (P<0.01. Although it is necessary to conduct further validation study using the other virus strains, MSD-ELISAs could be appropriate as a method to replace IS-ELISA for FMD antigen detection.

  18. Evaluation of monoclonal antibody-based sandwich direct ELISA (MSD-ELISA) for antigen detection of foot-and-mouth disease virus using clinical samples.

    Science.gov (United States)

    Morioka, Kazuki; Fukai, Katsuhiko; Sakamoto, Kenichi; Yoshida, Kazuo; Kanno, Toru

    2014-01-01

    A monoclonal antibody-based sandwich direct ELISA (MSD-ELISA) method was previously developed for foot-and-mouth disease (FMD) viral antigen detection. Here we evaluated the sensitivity and specificity of two FMD viral antigen detection MSD-ELISAs and compared them with conventional indirect sandwich (IS)-ELISA. The MSD-ELISAs were able to detect the antigen in saliva samples of experimentally-infected pigs for a longer term compared to the IS-ELISA. We also used 178 RT-PCR-positive field samples from cattle and pigs affected by the 2010 type-O FMD outbreak in Japan, and we found that the sensitivities of both MSD-ELISAs were about 7 times higher than that of the IS-ELISA against each sample (P<0.01). In terms of the FMD-positive farm detection rate, the sensitivities of the MSD-ELISAs were about 6 times higher than that of the IS-ELISA against each farm (P<0.01). Although it is necessary to conduct further validation study using the other virus strains, MSD-ELISAs could be appropriate as a method to replace IS-ELISA for FMD antigen detection.

  19. A novel indirect ELISA for diagnosis of dengue fever.

    Science.gov (United States)

    Narayan, Rohan; Raja, Senthil; Kumar, Senthil; Sambasivam, Mohana; Jagadeesan, Raja; Arunagiri, Kavita; Krishnasamy, Kaveri; Palani, Gunasekaran

    2016-07-01

    Dengue fever (DF) is associated with significant morbidity and mortality in the tropical and sub-tropical regions of the world. Since there are no effective antiviral drugs for treatment, clinicians often rely on the accurate diagnosis of dengue fever to begin supportive therapy at early stages of the illness. The objective of this study was to develop an in-house dengue virus serotype 2 (DENV-2) non-structural protein- 5 (NS5) based indirect ELISA. DENV-2 was raised in Vero cells and the viral proteins were separated and subsequently the NS5 protein was eluted. Serum samples from primary and secondary dengue fever patients; and acute and convalescent samples from Japanese encephalitis (JE) and West Nile virus (WNV) cases were used to validate the ELISA. The assay was found to be 100 per cent specific in detecting DENV-2 specific antibodies from patient's serum. However, in terms of sensitivity, the assay could detect IgM antibodies only from 90 per cent of the primary dengue samples. The IgM/IgG ratio of the primary and secondary samples was 7.24 and 0.64, respectively. The results indicate that the DENV-2 NS5 ELISA is dengue group specific and can be used to differentiate dengue infection from other circulating Flavivirus infections. This NS5 ELISA can also be used to distinguish between primary and secondary dengue fever on the basis of IgM/IgG ratios. Further studies with larger sample sizes and different DENV serotypes are required to validate the ELISA.

  20. A novel indirect ELISA for diagnosis of dengue fever

    Directory of Open Access Journals (Sweden)

    Rohan Narayan

    2016-01-01

    Full Text Available Background & objectives: Dengue fever (DF is associated with significant morbidity and mortality in the tropical and sub-tropical regions of the world. Since there are no effective antiviral drugs for treatment, clinicians often rely on the accurate diagnosis of dengue fever to begin supportive therapy at early stages of the illness. The objective of this study was to develop an in-house dengue virus serotype 2 (DENV-2 non-structural protein- 5 (NS5 based indirect ELISA. Methods: DENV-2 was raised in Vero cells and the viral proteins were separated and subsequently the NS5 protein was eluted. Serum samples from primary and secondary dengue fever patients; and acute and convalescent samples from Japanese encephalitis (JE and West Nile virus (WNV cases were used to validate the ELISA. Results: The assay was found to be 100 per cent specific in detecting DENV-2 specific antibodies from patient′s serum. However, in terms of sensitivity, the assay could detect IgM antibodies only from 90 per cent of the primary dengue samples. The IgM/IgG ratio of the primary and secondary samples was 7.24 and 0.64, respectively. Interpretation & conclusions: The results indicate that the DENV-2 NS5 ELISA is dengue group specific and can be used to differentiate dengue infection from other circulating Flavivirus infections. This NS5 ELISA can also be used to distinguish between primary and secondary dengue fever on the basis of IgM/IgG ratios. Further studies with larger sample sizes and different DENV serotypes are required to validate the ELISA.

  1. Distinct Patterns of IgG and IgA against Food and Microbial Antigens in Serum and Feces of Patients with Inflammatory Bowel Diseases

    Science.gov (United States)

    Frehn, Lisa; Jansen, Anke; Bennek, Eveline; Mandic, Ana D.; Temizel, Ilknur; Tischendorf, Stefanie; Verdier, Julien; Tacke, Frank; Streetz, Konrad; Trautwein, Christian; Sellge, Gernot

    2014-01-01

    Background Inflammatory bowel disease (IBD) is associated with a defective intestinal barrier and enhanced adaptive immune responses against commensal microbiota. Immune responses against food antigens in IBD patients remain poorly defined. Methods IgG and IgA specific for food and microfloral antigens (wheat and milk extracts; purified ovalbumin; Escherichia coli and Bacteroides fragilis lysates; mannan from Saccharomyces cerevisiae) were analyzed by ELISA in the serum and feces of patients with Crohn's disease (CD; n = 52 for serum and n = 20 for feces), ulcerative colitis (UC; n = 29; n = 17), acute gastroenteritis/colitis (AGE; n = 12; n = 9) as well as non-inflammatory controls (n = 61; n = 39). Results Serum anti-Saccharomyces cerevisiae antibodies (ASCA) and anti-B. fragilis IgG and IgA levels were increased in CD patients whereas antibody (Ab) levels against E. coli and food antigens were not significantly different within the patient groups and controls. Subgroup analysis revealed that CD patients with severe diseases defined by stricturing and penetrating lesions have slightly higher anti-food and anti-microbial IgA levels whereas CD and UC patients with arthropathy have decreased anti-food IgG levels. Treatment with anti-TNF-α Abs in CD patients was associated with significantly decreased ASCA IgG and IgA and anti-E. coli IgG. In the feces specific IgG levels against all antigens were higher in CD and AGE patients while specific IgA levels were higher in non-IBD patients. Anti-food IgG and IgA levels did not correlate with food intolerance. Summary In contrast to anti-microbial Abs, we found only minor changes in serum anti-food Ab levels in specific subgroups of IBD patients. Fecal Ab levels towards microbial and food antigens show distinct patterns in controls, CD and UC patients. PMID:25215528

  2. Maternofetal transplacental transport of recombinant IgG antibodies lacking effector functions

    DEFF Research Database (Denmark)

    Mathiesen, Line; Nielsen, Leif K; Andersen, Jan Terje

    2013-01-01

    alloimmunity, which may be lethal. A novel strategy to control pathogenic antibodies would be administration of a non-destructive IgG antibody blocking antigen binding while retaining binding to FcRn. We report on two human IgG3 antibodies with a hinge deletion and a C131S point mutation (IgG3ΔHinge......) that eliminate complement activation and binding to all classical Fcγ receptors (FcγRs) and to C1q while binding to FcRn is retained. Additionally, one of the antibodies has a single point mutation in the Fc (R435H) at the binding site for FcRn (IgG3ΔHinge:R435H). We compared transplacental transport with wild...... type IgG1 and IgG3, and found transport across trophoblast-derived BeWo cells and ex vivo placenta perfusions with hierarchies as follows: IgG3ΔHinge:R435H>wild type IgG1≥IgG3ΔHinge, and IgG3ΔHinge:R435H=wild type IgG1=wild type IgG3>IgG3ΔHinge, respectively. Collectively, IgG3ΔHinge:R435H...

  3. Enrichment of total serum IgG4 in patients with pemphigus

    Science.gov (United States)

    Funakoshi, T.; Lunardon, L.; Ellebrecht, C.T.; Nagler, A.R.; O’Leary, C.E.; Payne, A.S.

    2013-01-01

    Summary Background Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are potentially fatal blistering diseases caused by autoantibodies targeting desmoglein (Dsg) adhesion proteins. Previous studies have shown an IgG4 > IgG1 predominance of anti-Dsg antibodies in pemphigus; however, no studies have examined total serum IgG4 levels in pemphigus. IgG4 is induced by chronic antigen stimulation, which could occur with persistent skin blistering and potentially elevate the total serum IgG4 relative to other IgG subclasses in patients with pemphigus. Objectives The primary aim of the study was to quantitate total and Dsg-specific IgG subclasses in patients with pemphigus. Methods IgG subclasses and Dsg-specific IgG1 and IgG4 were quantitated in patients with PV and PF, and in sera from age-matched controls using a subclass enzyme-linked immunosorbent assay. The effectiveness of IgG4 depletion in blocking IgG pathogenicity in PV was determined using a keratinocyte dissociation assay. Results Dsg-specific antibodies comprised a median of 7·1% and 4·2% of total IgG4 in patients with PV and PF, respectively, with eightfold and fourfold enrichment in IgG4 vs. IgG1. Total serum IgG4, but not other IgG subclasses, was enriched in patients with PV and PF compared with age-matched controls (P = 0·004 and P = 0·005, respectively). IgG4 depletion of PV sera reduced pathogenicity in a keratinocyte dissociation assay and showed that affinity-purified IgG4 is more pathogenic than other serum IgG fractions. Conclusions Dsg-specific autoantibodies are significantly enriched in IgG4, which may explain the enrichment of total serum IgG4 in some patients with pemphigus. By preferentially targeting autoimmune rather than beneficial immune antibodies, IgG4-targeted therapies may offer safer treatment options for pemphigus. PMID:22803659

  4. Diagnostic sensitivity of cutoff values of IgG4-positive plasma cell number and IgG4-positive/CD138-positive cell ratio in typical multiple lesions of patients with IgG4-related disease.

    Science.gov (United States)

    Mizushima, Ichiro; Yamada, Kazunori; Harada, Kenichi; Matsui, Shoko; Saeki, Takako; Kondo, Satoru; Takahira, Masayuki; Waseda, Yuko; Hamaguchi, Yasuhito; Fujii, Hiroshi; Yamagishi, Masakazu; Kawano, Mitsuhiro

    2018-03-01

    This study aimed to investigate the diagnostic sensitivity of the cutoff values of IgG4-positive plasma cell (PC) number and IgG4-positive/CD138-positive cell ratio proposed by the International consensus statement (ICS) on the pathology of IgG4-related disease (IgG4-RD) in typical multiple lesions of patients with IgG4-RD. We evaluated IgG4-positive PC number and IgG4-positive/CD138-positive cell ratio in 39 samples from 18 IgG4-RD patients having more than two typical lesions of IgG4-RD. We evaluated 12 submandibular, 12 ophthalmic, six skin, five kidney, two pancreatic, and one bronchus and prostate lesion each in 18 patients with typical clinical, serological, and radiographic features. Concerning IgG4 + PC number per high-power field, most ophthalmic (11/12), kidney (5/5), pancreatic (2/2), and bronchial lesions (1/1) fulfilled the cutoff value of ICS, whereas many of the submandibular (6/12) and skin lesions (0/6) did not. In contrast to the absolute number, all lesions fulfilled the cutoff value of IgG4+/CD138 + cell ratio. In eight cases, only one or two lesions in the same patient fulfilled the cutoff value of ICS, while the others did not. These results suggest that ICS criteria have different sensitivities among the affected organs for the diagnosis of IgG4-RD.

  5. Prevalence of IgG varicella zoster virus antibodies in the Kuikuro and Kaiabi indigenous communities in Xingu National Park, Brazil, before varicella vaccination Prevalência de anticorpos IgG contra o vírus varicela zoster nas aldeias indígenas Kuikuro e Kaiabi do Parque Nacional do Xingu, Brasil, antes da vacinação contra varicela

    Directory of Open Access Journals (Sweden)

    Manuel Mindlin Lafer

    2005-06-01

    Full Text Available The purpose of the study was to estimate the prevalence of IgG antibodies against varicella zoster virus (VZV in the two most populated indigenous ethnic groups from Xingu Indigenous National Park, in Brazil, prior to the introduction of vaccination against the disease, and to determine the positive and the negative predictive values of a history of varicella infection. In 2001, 589 inhabitants of two Kuikuro villages and three Kaiabi villages were evaluated and provided information concerning previous varicella infection. An indirect immunosorbent assay (ELISA to detect IgG anti-VZV antibodies was performed in 224 blood samples - volunteer selection had no interference of anamnesis. IgG prevalence was 80.8% (95% Confidence Interval: 76% - 86%. The seroepidemiology of varicella in Xingu National Park prior to varicella vaccine introduction was comparable to the Brazilian national seroprevalence described in the literature, and so were the positive (98% and the negative predictive value (41% of the referred history.O objetivo do estudo foi aferir a prevalência de anticorpos IgG contra o Vírus Varicela-Zoster (VVZ nos dois grupos étnicos indígenas mais povoados do Parque Nacional Indígena do Xingu, Brasil, antes da introdução da vacinação contra a doença, e determinar os valores preditivos positivo e negativo da história de infecção de varicela. Em 2001, 589 habitantes de duas aldeias Kuikuro e três aldeias Kaiabi foram avaliados e forneceram dados referentes à infecção prévia por varicela. Um ensaio imunoenzimático indireto (ELISA foi realizado em 224 amostras de sangue para detectar anticorpos IgG anti-VVZ - a seleção de voluntários não teve interferência da anamnese. A prevalência de IgG foi de 80,8% (Intervalo de Confiança de 95%: 76% - 86%. A soroepidemiologia de varicela no Parque Nacional do Xingu antes da introdução da vacina foi comparável à soroprevalência nacional descrita na literatura, assim como os

  6. E.L.I.S.A. en coccidioidomicosis humana E.L.I.S.A. in human coccidioidomycosis

    Directory of Open Access Journals (Sweden)

    Iris Nora Tiraboschi

    1991-08-01

    Full Text Available Se realizó E.L.I.S.A. con exoantígeno de Coccidioides immitis para la detectión de anticuerpos, en 67 sueros humanos diluidos 1/1000, 1/2000, 1/4000 y 1/8000. De los 18 sueros de enfermos de coccidioidomicosis comprobada por examen directo, cultivo y/o histología, 5 fueron negativos, en otros 13 fueron positivos en una o varias diluciones. 3/26 sueros de personas sanas, coccidioidino positivas, fueron positivos en títulos de 1/1000 y el resto no tuvo anticuerpos detectables. No presentaron reacciones positivas ninguno de los sueros controles de personas sanas, pero sí lo hicieron 4/8 pacientes con otras micosis. Se concluye que E.L.I.S.A. es útil para la detección de mínimas cantidades de anticuerpos o en sueros que no pueden ser procesados por fijación de complemento. No es recomendable el uso de la técnica en forma aislada por al presencia de reacciones cruzadas.An E.L.I.S.A. test for antibody detection, with an exo-antigen of Coccidioides immitis was standardized in 67 humans sera diluited in 1/1000, 1/2000, 1/4000 and 1/8000. Eightheen sera from mycologically proved cases of coccidioidomycosis were studied: 5 were negative and 13 were positive in some dilutions. 3/26 sera of healthy persons who presented positive skin tests with coccidioidin were positive and the other 23 sera did not have positive reactions. None of the 15 sera of healthy human exhibited positive E.L.I.S.A. Serum samples of 8 patients suffering other deep mycosis were studied, 4 of them presented cross-reactions in E.L.I.S.A. tests. E.L.I.S.A. test seems to be a useful Serologic technique for antibody detection in anticomplementary serum samples or when a low concentration of antibodies should be detected. As it is very sensitive, cross-reactions with other mycoses are frequent, thus the use other more specific serologic technique together E.L.I.S.A. is recommended.

  7. [Standardization of Dot-ELISA for detection of anti-Trypanosoma cruzi antibodies, compared to ELISA and Western blot].

    Science.gov (United States)

    Cervantes-Landín, Alejandra Yunuen; Martínez-Martínez, Ignacio; Reyes, Pedro A; Shabib, Muslim; Espinoza-Gutiérrez, Bertha

    2014-01-01

    Chagas disease is considered endemic of Latin America. Because of migration of people from this region to non-endemic areas, such as the United States, Canada and Europe, it has become a major health problem. There are parasitology and serology tests for its diagnosis, but only the latter are useful during the chronic phase. Most of these tests require expensive equipment, which make them also inaccessible for laboratories in endemic areas. In the present work we standardize Dot-ELISA as a diagnostic test for Trypanosoma cruzi infection, since it is an easy, inexpensive and an accessible test. A total of 360 samples were tested: 96 sera from Chagas patients and 153 from healthy people; 40 blood samples spots collected and eluted from filter paper were also tested, as well as 71 serum samples of patients with non-related infections. Sensitivity, specificity and kappa index of Dot-ELISA test were calculated, in order to determine a correlation value of this technique compared to ELISA and Western blot that are already being used for diagnosis. Dot-ELISA obtained 97% sensitivity and 89% specificity, since it showed cross-reaction mainly with Leishmania spp., and a kappa index of 0,79. Dot-ELISA results correlate well with other tests that are already being used for diagnosis of Chagas disease. As it is easy and inexpensive, it may be useful as an additional diagnostic test or for field studies. Copyright © 2013 Elsevier España, S.L. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  8. Relação entre o diagnóstico sorológico (ELISA e a gravidade da tuberculose pulmonar na infância

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    Sant'Anna Clemax Couto

    2001-01-01

    Full Text Available Foram estudadas 48 crianças de 0 a 13 anos através da realização do ensaio imunoenzimático ligado a enzima (ELISA para pesquisa de anticorpos da classe IgG antiPPD, visando estabelecer correlação entre a resposta imune humoral medida pela sorologia e a gravidade da tuberculose, segundo formas radiológicas (leve, moderada e grave. A amostra foi composta de 29 crianças com tuberculose e 19 sem tuberculose comunicantes de tuberculose. Os valores médios (medianas da densidade óptica do teste ELISA foram, respectivamente: 0,098 na forma gânglio-pulmonar (leve, 0,092 na forma pneumônica (moderada e 0,134 na tuberculose miliar (grave. Nas crianças não tuberculosas com radiografia de tórax normal, o ELISA foi igual a 0,020. Os achados evidenciam valores mais elevados do teste sorológico relacionados à maior gravidade da doença (p= 0,0007.

  9. Characterization of the Native and Denatured Herceptin by ELISA and QCM using a High-Affinity Single Chain Fragment Variable (scFv) Recombinant Antibody

    Science.gov (United States)

    Shang, Yuqin; Mernaugh, Ray

    2012-01-01

    Herceptin/Trastuzumab is a humanized IgG1κ light chain antibody used to treat some forms of breast cancer. A phage-displayed recombinant antibody library was used to obtain an scFv (designated 2B4) to a linear synthetic peptide representing Herceptin’s heavy chain CDR3. ELISAs and piezoimmunosensor/quartz crystal microbalance (QCM) assays were used to characterize 2B4-binding activity to both native and heat denatured Herceptin. The 2B4 scFv specifically bound to heat denatured Herceptin in a concentration dependent manner over a wide (35–220.5 nM) dynamic range. Herceptin denatures and forms significant amount of aggregates when heated. UV-Vis characterization confirms that Herceptin forms aggregates as the temperature used to heat Herceptin increases. QCM affinity assay shows that binding stoichiometry between 2B4 scFv and Herceptin follows a 1:2 relationship proving that 2B4 scFv binds strongly to the dimers of heat denatured Herceptin aggregates and exhibits an affinity constant of 7.17 × 1013 M−2. The 2B4-based QCM assay was more sensitive than the corresponding ELISA. Combining QCM with ELISA can be used to more fully characterize non-specific binding events in assays. The potential theoretical and clinical implications of these results and the advantages of using QCM to characterize human therapeutic antibodies in samples are also discussed. PMID:22934911

  10. Spectrum of Disorders Associated with Elevated Serum IgG4 Levels Encountered in Clinical Practice

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    Jay H. Ryu

    2012-01-01

    Full Text Available IgG4-related disease (IgG4-RD is a recently described systemic fibroinflammatory disease associated with elevated circulating levels of IgG4 and manifests a wide spectrum of clinical presentations. Although serum IgG4 level has been described to be the most sensitive and specific laboratory test for the diagnosis of IgG4-RD, it is recognized that an elevated serum IgG4 level can be encountered in other diseases. In this study, we sought to identify the frequency of IgG4-RD and other disease associations in patients with elevated serum IgG4 levels seen in clinical practice. Among 3,300 patients who underwent IgG subclass testing over a 2-year period from January 2009 to December 2010, 158 (4.8% had an elevated serum IgG4 level (>140 mg/dL. IgG4 subclass testing was performed for evaluation of suspected IgG4-RD or immunodeficiency. Twenty-nine patients (18.4% had definite or possible IgG4-RD. Among those patients without IgG4-RD, a broad spectrum of biliary tract, pancreatic, liver, and lung diseases, as well as systemic vasculitis, was diagnosed. We conclude that patients with elevated serum IgG4 levels encountered in clinical practice manifest a wide array of disorders, and only a small minority of them has IgG4-RD.

  11. The role of IgG in immediate-type hypersensitivity

    NARCIS (Netherlands)

    van der Zee, J. S.; Aalberse, R. C.

    1991-01-01

    Two opposed modes of action have been attributed to immunoglobulin G (IgG) in immediate-type allergy. On the one hand, a small fraction of IgG may have anaphylactic properties. On the other hand, IgG antibodies induced by hyposensitization therapy are considered to act as allergen blocking

  12. Human IgG subclass antibodies to the 19 kilodalton carboxy ...

    African Journals Online (AJOL)

    Human IgG subclass antibodies to the 19 kilodalton carboxy terminal fragment of Plasmodium Falciparum merozoite surface protein 1 (MSP1 19 ) and predominance of the MAD20 allelic ... Results: Both the prevalence and the mean concentration of serum IgG1 , and to a lesser extent IgG3, antibodies increased with age.

  13. On the Perplexingly Low Rate of Transport of IgG2 across the Human Placenta

    NARCIS (Netherlands)

    Einarsdottir, Helga K.; Stapleton, Nigel M.; Scherjon, Sicco; Andersen, Jan Terje; Rispens, Theo; van der Schoot, C. Ellen; Vidarsson, Gestur

    2014-01-01

    The neonatal receptor, FcRn, mediates both serum half-life extension as well as active transport of maternal IgG to the fetus during pregnancy. Therefore, transport efficiency and half-life go hand-in-hand. However, while the half-life of the human IgG2 subclass is comparable to IgG1, the placental

  14. The role of IgG antibodies in allergy and immunotherapy

    NARCIS (Netherlands)

    Aalberse, R.

    2011-01-01

    In specific immunotherapy (SIT), a beneficial response is associated with an increase in allergen-specific IgG(4) . This does not indicate that IgE-producing B cells have switched to IgG(4) production, because in human DNA, IgE is downstream from IgG(4) . Thus, by conventional switching, B cells

  15. Evaluation of bio-conjugation methods for obtaining of synthetic positive control for IgM-capture ELISA

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    O. Y. Galkin

    2014-10-01

    Full Text Available The enzyme-linked immunosorbent assay (ELISA is the most informative and versatile method of serological diagnostics. The possibility of detecting by ELISA specific antibodies of different classes allow to differentiate primary infectious process and its remission, exacerbation and chronic disease (differential diagnosis. This approach is implemented in the methodology for evaluation of patients for the presence of humoral immune response to TORCH-infections pathogens (toxoplasmosis, rubella, cytomegalovirus, herpes simplex viruses’ infections, and some others. Therefore, testing for presence of specific IgG and IgM antibodies against TORCH-infections pathogens in blood serum is an important element of motherhood and childhood protection. The essential problem in the production of IgM-capture ELISA diagnostic kits is obtaining of positive control. The classic version of positive control is human blood serum (plasma containing specific antibodies. But specific IgM-positive sera are insignificant raw materials. This fact can seriously restrict the production of diagnostic kits, especially in the event of large-scale production. We have suggested the methodological approach to using of synthetic positive controls in IgM-capture ELISA kits based on conjugate of normal human IgM and monoclonal antibodies against horseradish peroxidase. It is found that this task can be fulfilled by means of NHS ester-maleimide-mediated conjugation (by sulfosuccinimidyl-4-(N-maleimidomethylcyclohexane-1-carboxylate, reductive amination-mediated conjugation (by sodium periodate and glutaraldehyde-mediated conjugation. It was found that conjugates of normal human IgM and monoclonal antibodies against horseradish peroxidase obtained using NHS ester-mediated maleimide conjugation and periodate method were similar by molecular weight, whereas conjugate synthesized by glutaraldehyde method comprised at least three types of biopolymers with close molecular weight. It was

  16. Efficacy demonstration of tetanus vaccines by double antigen ELISA.

    Science.gov (United States)

    Rosskopf, U; Noeske, K; Werner, E

    2005-09-01

    This paper describes a double antigen ELISA (DAE) for rapid, specific and reliable assessment of the antitetanus immune status of horses and sheep. Compared with the indirect ELISA, the double antigen ELISA has the advantage of species-independent testing of sera. Thanks to its test design, it is more specific since the detected antibodies are forced to bind tetanus toxoid twice. In addition, it is very sensitive to tetanus antibodies, enabling the detection of low antibody titres, in range which is relevant for the assessment of the protective status (tetanus toxin neutralising antibodies). The detection limit of the DAE for tetanus antibodies is in the order of 10(-4) EU/ml. A comparison of in vitro results of individual sera with in vivo titres showed that horse sera with titres of 0.04 and 0.05 EU/ml in the DAE showed titres of > 0.05 IU and 0.034 IU/ml respectively during in vivo testing thus indicating good agreement. For tested sheep sera which were rated > 0.05 IU/ml in vivo, the corresponding titre in the DAE was 0.24 EU/ml. Clear tetanus antitoxin establishment of protective ELISA limits requires further comparative examination of sera with low titres (tetanus vaccines ad us. vet. As a consequence, the toxin neutralisation test (still being the standard method of choice for quantifying tetanus toxin neutralising antitoxin titres) could be replaced, since it requires too great a number of animals per test and involves considerable suffering for the animals. The test described here reduces the use of mice and guinea pigs within vaccine efficacy testing. In addition, it involves less exposure of the laboratory personnel to toxin.

  17. Determination of specific IgG antibody by crossed radioimmunoelectrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Nordvall, S.L. (Dept. of Paediatrics, University Hospital, Uppsala, Sweden); Uhlin, T.; Einarsson, R. (Allergy Research, Pharmacia Diagnostics AB, Uppsala, Sweden)

    1983-01-01

    A crossed radioimmunoelectrophoretic method was developed for detection of honey bee venom specific IgG antibodies in patient sera. At the serum concentration 1/200 the contrast between specific binding and backgroud was the most favourable. The detection limit was fairly low, approximately 30 kU/l(IgG RAST units). A reference system based on the reference kits in Phadebas IgG-RAST was elaborated.

  18. Determination of specific IgG antibody by crossed radioimmunoelectrophoresis

    International Nuclear Information System (INIS)

    Nordvall, S.L.; Uhlin, T.; Einarsson, R.

    1983-01-01

    A crossed radioimmunoelectrophoretic method was developed for detection of honey bee venom specific IgG antibodies in patient sera. At the serum concentration 1/200 the contrast between specific binding and backgroud was the most favourable. The detection limit was fairly low, approximately 30 kU/l(IgG RAST units). A reference system based on the reference kits in Phadebas IgG-RAST was elaborated. (author)

  19. Genetic and infectious profiles influence cerebrospinal fluid IgG abnormality in Japanese multiple sclerosis patients.

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    Satoshi Yoshimura

    Full Text Available BACKGROUND: Abnormal intrathecal synthesis of IgG, reflected by cerebrospinal fluid (CSF oligoclonal IgG bands (OBs and increased IgG index, is much less frequently observed in Japanese multiple sclerosis (MS cohorts compared with Western cohorts. We aimed to clarify whether genetic and common infectious backgrounds influence CSF IgG abnormality in Japanese MS patients. METHODOLOGY: We analyzed HLA-DRB1 alleles, and IgG antibodies against Chlamydia pneumoniae, Helicobacter pylori, Epstein-Barr virus nuclear antigen (EBNA, and varicella zoster virus (VZV in 94 patients with MS and 367 unrelated healthy controls (HCs. We defined CSF IgG abnormality as the presence of CSF OBs and/or increased IgG index (>0.658. PRINCIPAL FINDINGS: CSF IgG abnormality was found in 59 of 94 (62.8% MS patients. CSF IgG abnormality-positive patients had a significantly higher frequency of brain MRI lesions meeting the Barkhof criteria compared with abnormality-negative patients. Compared with HCs, CSF IgG abnormality-positive MS patients showed a significantly higher frequency of DRB1 1501, whereas CSF IgG abnormality-negative patients had a significantly higher frequency of DRB1 0405. CSF IgG abnormality-positive MS patients had a significantly higher frequency of anti-C. pneumoniae IgG antibodies compared with CSF IgG abnormality-negative MS patients, although there was no difference in the frequency of anti-C. pneumoniae IgG antibodies between HCs and total MS patients. Compared with HCs, anti-H. pylori IgG antibodies were detected significantly less frequently in the total MS patients, especially in CSF IgG abnormality-negative MS patients. The frequencies of antibodies against EBNA and VZV did not differ significantly among the groups. CONCLUSIONS: CSF IgG abnormality is associated with Western MS-like brain MRI features. DRB1 1501 and C. pneumoniae infection confer CSF IgG abnormality, while DRB1 0405 and H. pylori infection are positively and negatively

  20. Dot-ELISA for the diagnosis of neurocysticercosis Dot-ELISA no diagnóstico da neurocisticercose

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    Rakhi Biswas

    2004-10-01

    Full Text Available The aim of the present study was to standardize and evaluate dot-Enzyme linked immunosorbent assay (Dot-ELISA, a simple and rapid test for the detection of cysticercus antibodies in the serum for the diagnosis of neurocysticercosis (NCC. The antigen used in the study was a complete homogenate of Cysticercus cellulosae cysts obtained from infected pigs and dotted on to nitrocellulose membrane. Test sera were collected from the patients of NCC, and control sera from patients with other diseases and healthy students and blood donors of the Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER Hospital, Pondicherry, during a study period from 2001 to 2003. Dot-ELISA detected antibodies in 14 of 25 (56% in clinically suspected cases of NCC, 13 of 23 (56.5% in CT/MRI proven cases of NCC and 2 of 25 (8% each in non-cysticercal CNS infection controls and healthy controls. The test showed a sensitivity of 56.25%, specificity of 92%, positive predictive value of 87.09%, and negative predictive value of 70.76%. Results of the present study shows that the Dot-ELISA as a simple test can be used in the field or poorly equipped laboratories for diagnosis of NCC .O objetivo do presente estudo foi estandardizar e avaliar o Dot-ELISA, um teste simples e rápido para detectar anticorpos de cisticercos no soro para diagnóstico da neurocisticercose (NCC. O antígeno usado no estudo foi um homogenizado completo de cistos de Cysticercus cellulosae obtidos de porcos infectados e marcados sobre a membrana de nitrocelulose. Os soros testados foram coletados de pacientes com NCC e os soros controle de pacientes com outras doenças e estudantes saudáveis e doadores e sangue do "Jawaharlal Institute of Postgraduate Medical Education and Research Hospital", em Pondicherry, durante o período de estudo de 2001 a 2003. Dot-Elisa detectou anticorpos em 14 de 25 (56% casos suspeitos de NCC, em 13 de 23 (56,5% em CT/MRI casos provados de NCC e em 2 de 25

  1. Quantitative sandwich ELISA for the determination of fish in foods.

    Science.gov (United States)

    Faeste, Christiane K; Plassen, Christin

    2008-01-01

    Allergy to fish represents one of the most prevalent causes for severe food-allergic reactions. Therefore, food authorities in different countries have implemented mandatory labeling of fish in pre-packed foods. Detection of fish proteins in food has previously been based on the use of patient serum. In the present study, a novel sandwich enzyme-linked immunosorbent assay (ELISA) for the quantitation of fish in food matrixes has been developed and validated, using a polyclonal rabbit anti-cod parvalbumin antibody for capture and a biotinylated conjugate of the same antibody for detection. By employing the ubiquitous muscle protein parvalbumin as target the method succeeds to detect a variety of fish. However, the ELISA is specific for fish and does not cross-react with other species. Recoveries ranged from 68-138% in typical food matrixes, while the intra- and inter-assay precisions were parvalbumin ELISA with a limit of detection of 0.01 mg parvalbumin/kg food, about 5 mg fish/kg food, seems sufficient to detect fish protein traces in foods at levels low enough to minimize the risk for fish allergic consumers.

  2. Difference of Clinical and Radiological Characteristics According to Radioiodine Avidity in Pulmonary Metastases of Differentiated Thyroid Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Dohoon; Jung, Jihoon; Song, Seung Hyun; Kim, Choonyoung; Hong, Chae Moon; Jeong, Shin Young; Lee, Sangwoo; Lee, Jaetae; Ahn, Byeongcheol [Kyungpook National Univ., Daegu (Korea, Republic of)

    2014-03-15

    To evaluate differences in clinical, radiological and laboratory findings between pulmonary metastasis with and without radioiodine avidity in thyroidectomized differentiated thyroid cancer (DTC) patients with pulmonary metastasis who underwent high-dose I-131 treatment. A total of 105 DTC patients with pulmonary metastasis (age, 48.7±16.8 years; women/men, 78/27) were included. Clinical characteristics, chest computed tomography (CT), F-18 fluorodeoxyglucose positron emission tomography (F-18 FDG PET)/CT and thyroid-stimulating hormone (TSH)-stimulated serum thyroglobulin (s-Tg) level were compared between patients with and without radioiodine uptake in metastatic lung lesions. The response to I-131 treatment was evaluated with follow-up study. Eighty-nine patients (84.8 %, whole-body scan positive [WBSP] group) showed radioiodine uptake at pulmonary metastasis on post I-131 treatment whole-body scan (WBS) and 16 patients (15.2 %, WBS negative [WBSN] group) did not show uptake at pulmonary lesions on the WBS. Ninety percent and 87 % of the WBSP group had visible metastatic lesions on CT and F-18 FDG PET/CT; however, all of the patients in the WBSN group showed lesions on CT and F-18 FDG PET/CT. In seven (6.7 %) of 105 patients, CT and F-18 FDG PET/CT could not detect pulmonary lesions, which were diagnosed by post I-131 treatment WBS. Complete disease remission was achieved in six (5.7 %) patients and all of them were in the WBSP group. Metastatic lesion was not visualized on chest CT or F-18 FDG PET/CT in 6.7 % of DTC patients with pulmonary metastasis and the lesion was visualized only on post I-131 treatment WBS. Complete remission was achieved in 5.7 % of DTC patients with pulmonary metastasis and the cured metastases were non-visualizing or micronodular lesions on chest CT and demonstrated radioiodine avidity on post I-131 treatment WBS.

  3. Overlap of highly FDG-avid and FMISO hypoxic tumor subvolumes in patients with head and neck cancer.

    Science.gov (United States)

    Mönnich, David; Thorwarth, Daniela; Leibfarth, Sara; Pfannenberg, Christina; Reischl, Gerald; Mauz, Paul-Stefan; Nikolaou, Konstantin; la Fougère, Christian; Zips, Daniel; Welz, Stefan

    2017-11-01

    PET imaging may be used to personalize radiotherapy (RT) by identifying radioresistant tumor subvolumes for RT dose escalation. Using the tracers [ 18 F]-fluorodeoxyglucose (FDG) and [ 18 F]-fluoromisonidazole (FMISO), different aspects of tumor biology can be visualized. FDG depicts various biological aspects, e.g., proliferation, glycolysis and hypoxia, while FMISO is more hypoxia specific. In this study, we analyzed size and overlap of volumes based on the two markers for head-and-neck cancer patients (HNSCC). Twenty five HNSCC patients underwent a CT scan, as well as FDG and dynamic FMISO PET/CT prior to definitive radio-chemotherapy in a prospective FMISO dose escalation study. Three PET-based subvolumes of the primary tumor (GTV prim ) were segmented: a highly FDG-avid volume V FDG , a hypoxic volume on the static FMISO image acquired four hours post tracer injection (V H ) and a retention/perfusion volume (V M ) using pharmacokinetic modeling of dynamic FMISO data. Absolute volumes, overlaps and distances to agreement (DTA) were evaluated. Sizes of PET-based volumes and the GTV prim are significantly different (GTV prim >V FDG >V H >V M ; p DTAs are small (mean coverage 74.4%, mean DTA 1.4 mm). Coverage of V M is less pronounced. With respect to V FDG and V H , the mean coverage is 48.7% and 43.1% and the mean DTA is 5.3 mm and 6.3 mm, respectively. For two patients, DTAs were larger than 2 cm. Hypoxic subvolumes from static PET imaging are typically covered by or in close proximity to highly FDG-avid subvolumes. Therefore, dose escalation to FDG positive subvolumes should cover the static hypoxic subvolumes in most patients, with the disadvantage of larger volumes, resulting in a higher risk of dose-limiting toxicity. Coverage of subvolumes from dynamic FMISO PET is less pronounced. Further studies are needed to explore the relevance of mismatches in functional imaging.

  4. Avidity Binding of Human Adenovirus Serotypes 3 and 7 to the Membrane Cofactor CD46 Triggers Infection

    Science.gov (United States)

    Trinh, Hung V.; Lesage, Guillaume; Chennamparampil, Venus; Vollenweider, Benedikt; Burckhardt, Christoph J.; Schauer, Stefan; Havenga, Menzo; Greber, Urs F.

    2012-01-01

    The species B human adenoviruses (HAdVs) infect cells upon attaching to CD46 or desmoglein 2 (DSG-2) by one or several of their 12 fiber knob trimers (FKs). To test whether DSG-2 and CD46 simultaneously serve as virus receptors for adenovirus type 3 (Ad3), we performed individual and combined CD46/DSG-2 loss-of-function studies in human lung A549 and 16HBE14o cells. Our results suggest that in these cells, DSG-2 functions as a major attachment receptor for Ad3, whereas CD46 exerts a minor contribution to virus attachment and uptake in the range of ∼10%. However, in other cells the role of CD46 may be more pronounced depending on, e.g., the expression levels of the receptors. To test if avidity allows Ad3/7 to use CD46 as a receptor, we performed gain-of-function studies. The cell surface levels of ectopically expressed CD46 in CHO or human M010119 melanoma cells lacking DSG-2 positively correlated with Ad3/7 infections, while Ad11/35 infections depended on CD46 but less on CD46 levels. Antibody-cross-linked soluble CD46 blocked Ad3/7/11/35 infections, while soluble CD46 alone blocked Ad11/35 but not Ad3/7. Soluble Ad3/7-FKs poorly inhibited Ad3/7 infection of CHO-CD46 cells, illustrating that Ad3/7-FKs bind with low affinity to CD46. This was confirmed by Biacore studies. Ad3/7-FK binding to immobilized CD46 at low density was not detected, unlike that of Ad11/35-FK. At higher CD46 densities, however, Ad3/7-FK bound to CD46 with only 15-fold-higher dissociation constants than those of Ad11/35-FK. These data show that an avidity mechanism for Ad3/7 binding to CD46 leads to infection of CD46-positive cells. PMID:22130529

  5. Transfection of an expressive construct including IgG1 and Fv1 genes in ovary cell line for infliximab expression

    Directory of Open Access Journals (Sweden)

    Zohreh Sarabinejad

    2016-04-01

    Full Text Available Background: Infeliximab is a form of chimeric antibody which neutralizes the most important inflammatory cytokine, TNF-a, in inflammatory disorders. The aim of current study was to pilot expression of chimeric infliximab in Chinese Hamster ovary (CHO cells. Methods: In this research study, pVITRO2-neo-mcs vector that consist of infliximab light chain and heavy chain was used to transform into the E.coli by CaCl2 method. The plasmid was then purified and transfected to cultured CHO cells by Lipofectamine 2000® (Invitrogen GmbH, Germany. Transfected cells were selected upon G-418 treatment after 2 weeks and the level of expression, based on standard curve, was measured using IgG ELISA kit after 48 hours for each clone. High level expressed clone was then cultured in roller bottles and recombinant chimeric product was purified by protein A affinity chromatography. The purity of the product was analyzed by 10% gel SDS-PAGE from eluted samples. The efficacy of the purification was analyzed by ELISA before and after purification step. This article is a master's student thesis from February 2015 to August 2016 in pharmaceutical technology development center, Tehran University of Medical Sciences, Tehran, Iran. Results: The purified plasmid was analyzed on 2% agarose gel. After selective pressure of G-418, 10 stable transfect clones were assessed for infliximab secretion by IgG ELISA kit at 450 nm. The maximum and minimum expression which detected by ELISA were 23 ng/ml and 6 ng/ml, respectively. The band width of infliximab fraction during purification procedure was observed at 0.7-0.8 min. The efficiency of the purification by ELISA was 70%. On SDS-PAGE analysis, two bands, 25 and 50 kDa, respect to light and heavy chains of Infliximab, was confirmed the expression of recombinant protein. Conclusion: In the current study, the construct for infliximab monoclonal antibody production was designed using genetic engineering techniques and the expression

  6. IgG4 subclass antibodies impair antitumor immunity in melanoma.

    Science.gov (United States)

    Karagiannis, Panagiotis; Gilbert, Amy E; Josephs, Debra H; Ali, Niwa; Dodev, Tihomir; Saul, Louise; Correa, Isabel; Roberts, Luke; Beddowes, Emma; Koers, Alexander; Hobbs, Carl; Ferreira, Silvia; Geh, Jenny L C; Healy, Ciaran; Harries, Mark; Acland, Katharine M; Blower, Philip J; Mitchell, Tracey; Fear, David J; Spicer, James F; Lacy, Katie E; Nestle, Frank O; Karagiannis, Sophia N

    2013-04-01

    Host-induced antibodies and their contributions to cancer inflammation are largely unexplored. IgG4 subclass antibodies are present in IL-10-driven Th2 immune responses in some inflammatory conditions. Since Th2-biased inflammation is a hallmark of tumor microenvironments, we investigated the presence and functional implications of IgG4 in malignant melanoma. Consistent with Th2 inflammation, CD22+ B cells and IgG4(+)-infiltrating cells accumulated in tumors, and IL-10, IL-4, and tumor-reactive IgG4 were expressed in situ. When compared with B cells from patient lymph nodes and blood, tumor-associated B cells were polarized to produce IgG4. Secreted B cells increased VEGF and IgG4, and tumor cells enhanced IL-10 secretion in cocultures. Unlike IgG1, an engineered tumor antigen-specific IgG4 was ineffective in triggering effector cell-mediated tumor killing in vitro. Antigen-specific and nonspecific IgG4 inhibited IgG1-mediated tumoricidal functions. IgG4 blockade was mediated through reduction of FcγRI activation. Additionally, IgG4 significantly impaired the potency of tumoricidal IgG1 in a human melanoma xenograft mouse model. Furthermore, serum IgG4 was inversely correlated with patient survival. These findings suggest that IgG4 promoted by tumor-induced Th2-biased inflammation may restrict effector cell functions against tumors, providing a previously unexplored aspect of tumor-induced immune escape and a basis for biomarker development and patient-specific therapeutic approaches.

  7. Modification the ELISA Kit for diagnosis of “Pseudomonas aeruginosa and comparing it with ordinary ELISA kit”

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    Taghreed A. Mohammad

    2017-07-01

    Full Text Available The first aim of the present study was to diagnosis Pseudomonas  aeruginosa by many tests. This study consisted "200 patients " who suffered from burn wound and compare with 100 health individuals (male and female as a control group, Vitek test was used to diagnose  118 (87 "local isolate  ATCC 15692" with 31 other isolate of Pseudomonas  aeruginosa  ((ATCC 15690, ATCC 15688  from 200 samples which were taken from burn patients. This result was similar to Analytical profile index ( API   test (118 isolates of  P. aeruginosa with 82 isolations of  other bacteria. Then the detection P. aeruginosa isolate  ATCC 15692 by new ELISA Technique and comparing its with modify the ordinary ELISA kit.

  8. Immuno-epidemiology of human Schistosoma haematobium infection: preferential IgG3 antibody responsiveness to a recombinant antigen dependent on age and parasite burden

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    Fernandez Cecilia

    2006-06-01

    Full Text Available Abstract Background Schistosomiasis is a major parasitic disease affecting over 200 million people in the developing world with a further 400 million people at risk of infection. The aim of this study was to identify a single antigen from adult Schistosoma haematobium worms and subsequently use this antigen to study the development of schistosome-acquired immunity in a human population. Methods The full-length cDNA sequence of a S. haematobium protein, a putative orthologue of the S. mansoni tegumental antigen Sm13, was obtained from a cDNA library of adult S. haematobium worms and named Sh13 following a small-scale expressed sequence tags (EST project. The recombinant Sh13 protein expressed in E. coli, was used to investigate immuno-epidemiological patterns in 147 Zimbabweans (7–18 years old exposed to S. haematobium. Results Sequence analysis of the full-length cDNA sequence of the S. haematobium protein Sh13, indicated that the protein has an N-terminal signal peptide and encodes an 85-amino acid mature protein with a highly conserved predicted transmembrane domain (86 % identity with the S. mansoni tegumental antigen Sm13. The recombinant Sh13 protein was used in ELISA assays to determine the reactivity of sera from the study participants. Antibody responses against Sh13 were predominantly IgG3 isotype compared to responses against crude worm antigens which were predominantly IgG1 and IgG4. The relationship between anti-Sh13 IgG3 levels and infection intensity varied significantly with host age. The youngest children (7–10 years old had relatively low levels of both infection and anti-Sh13 IgG3. In older children (11–12 years old rising infection levels were accompanied by a significant increase in anti-Sh13 IgG3 levels. Subsequently, infection intensity declined significantly in 13–18 year olds but levels of the antibody continued to rise. The changing relationship between infection intensity and anti-Sh13 IgG3 levels with host age

  9. [Detection the serum IgM and IgG specific for TORCH in 1307 women in the period of pre-pregnancy and pregnancy].

    Science.gov (United States)

    Qin, Wen; Hu, Da-chun; Pang, Hai-li

    2011-08-01

    The objective is to obtain the information of women in the period of prepregnancy and pregnancy infected by TORCH (Toxoplasma, Rabella Virus, Cytomegalovirus, and Herpes Simplex Virus) in Kunming for preventing fetus from infection by TORCH in uterus. The serum IgM and IgG specific for TORCH from 1307 women in the period of pre-pregnancy and pregnancy were measured with ELISA and The reagent is the product of virion/serion. The serum IgM specific for TORCH were detected in 2.83% of 1307 pregnant women for Toxoplasma, 2.37% for Rebulla Virus, 0.46% for Cytomegalovirus, 2.45% for Herpes Simplex Virus. The total positive rate of serum IgM specific for TORCH was 1.45%. The serum IgG specific for TORCH were detected in 3.98% of 1307 pregnant women for Toxoplasma, 72.3% for Rebulla Virus, 97.78% for Cytomegalovirus, 80.34% for Herpes Simplex Virus. The total positive rate of serum IgG specific for TORCH was 63.60%. There was no significant difference among the women with different pregnant situation in terms of the serum IgM and IgG specific for TORCH. There are some of women in the period of pre-pregnancy and pregnancy with current infection by TORCH in Kunming. It is recommended to screen for infection by TORCH in women in the period of pre-pregnancy and pregnancy to prevent fetus from infection by TORCH in uterus.

  10. Comparison of IgG and IgA Antibodies Titrations against Helicobacter Pylori in Urban and Rural Populations in Mazandaran Province

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    Mina Owrang

    2014-06-01

    Full Text Available Background & objective: The infection caused by Helicobacter pylori (H. pylori is one of the most common bacterial gastrointestinal diseases throughout the world. Based on the role of H. pylori in a variety of diseases such as gastrointestinal and lymphoma, present study is aimed to consider the concentration of IgA and IgG against H. pylori in both rural and urban populations and then its relationship with some demographic characteristics. Materials & Methods: In this descriptive study, 400 sera samples were collected from both genders at the Sari treatment- health center. After blood collection, the concentration of IgG and IgA against H. pylori was measured by ELISA kit. Results: There was no significant difference in antibodies titration between men and women. Approximately 18.5% of males and 16.5% of females were positive regarding to IgA and 70.2% of men and 66.7% of women were positive regarding to IgG. The mean of antibodies in rural populations (0.87±0.35 was significantly (p<0.001 higher than those in urban populations (0.78±0.41. The mean of antibodies in patients who had a history of gastrointestinal infection was significantly higher than others (p<0.05. Conclusion: Due to the high level of IgA and IgG antibodies in studied populations, especially in rural people, and lack of symptoms in patients, the screen of positive serologic populations can be helpful for the management and control of infections caused by H. pylori.

  11. High prevalence of IgG antibodies to Ebola virus in the Efé pygmy population in the Watsa region, Democratic Republic of the Congo.

    Science.gov (United States)

    Mulangu, Sabue; Borchert, Matthias; Paweska, Janusz; Tshomba, Antoine; Afounde, Afongenda; Kulidri, Amayo; Swanepoel, Robert; Muyembe-Tamfum, Jean-Jacques; Van der Stuyft, Patrick

    2016-06-10

    Factors related to the natural transmission of Ebola virus (EBOV) to humans are still not well defined. Results of previous sero-prevalence studies suggest that circulation of EBOV in human population is common in sub-Saharan Africa. The Efé pygmies living in Democratic Republic of the Congo are known to be exposed to potential risk factors of EBOV infection such as bush meat hunting, entry into caves, and contact with bats. We studied the pygmy population of Watsa region to determine seroprevalence to EBOV infection and possible risks factors. Volunteer participants (N = 300) aged 10 years or above were interviewed about behavior that may constitute risk factors for transmission of EBOV, including exposures to rats, bats, monkeys and entry into caves. Samples of venous blood were collected and tested for IgG antibody against EBOV by enzyme-linked immunosorbent assay (ELISA). The χ2-test and Fisher's exact test were used for the comparison of proportions and the Student's t-test to compare means. The association between age group and anti-EBOV IgG prevalence was analysed by a nonparametric test for trend. The prevalence of anti-EBOV IgG was 18.7 % overall and increased significantly with age (p = 0.023). No association was observed with exposure to risk factors (contacts with rats, bats, monkeys, or entry into caves). The seroprevalence of IgG antibody to EBOV in pygmies in Watsa region is among the highest ever reported, but it remains unclear which exposures might lead to this high infection rate calling for further ecological and behavioural studies.

  12. A novel lateral flow immunoassay for the rapid detection of anti-Dsg3 IgG serum autoantibodies in pemphigus vulgaris.

    Science.gov (United States)

    Schmidt, Thomas; Mauracher, Susanne; Bender, Lena; Greene, Brandon; Kurzhals, Jonas; Eming, Rüdiger; Dostatni, Ralf; Hertl, Michael

    2017-12-26

    Pemphigus vulgaris (PV) is a severe autoimmune blistering disease of the skin and mucous membranes. As autoantibodies play an essential role in the disease pathogenesis, the serological detection of anti-desmoglein 3 IgG represents a central tool in the diagnosis of the disease. In this study, we show the validation of a novel lateral flow immunoassay (LFIA) which rapidly detects anti-desmoglein 3 (Dsg3) IgG in human serum. In contrast to other diagnostic procedures, the assay is compact and simple to perform and delivers a fast "yes" or "no" answer within 10 minutes without additional hardware requirements for test evaluation. For validation, a blinded collection of 200 sera including 100 sera from 14 PV patients, 75 sera from 24 bullous pemphigoid patients and 25 sera from 6 patients with pemphigus foliaceus collected at different time points during disease was used. Presence or non-presence of anti-Dsg3 IgG within sera was confirmed using a commercially available Dsg3-ELISA. For qualitative evaluation, Dsg3-LFIA test results were assessed by two independent groups of human observers. Furthermore, quantitative evaluation using POCScan reader was applied. The Dsg3-LFIA demonstrated reliable test results with a sensitivity and specificity of 78.1% and 97.1%, respectively. Test results from POCScan and human observers showed a substantial agreement. The Dsg3-LFIA represents a new diagnostic tool for the immediate and reliable detection of anti-desmoglein 3 serum IgG autoantibodies that does not require additional hardware. Further prospective trials are warranted to validate the Dsg3 LFIA in pemphigus. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Seroprevalence study in forestry workers from eastern Germany using novel genotype 3- and rat hepatitis E virus-specific immunoglobulin G ELISAs.

    Science.gov (United States)

    Dremsek, Paul; Wenzel, Jürgen J; Johne, Reimar; Ziller, Mario; Hofmann, Jörg; Groschup, Martin H; Werdermann, Sandra; Mohn, Ulrich; Dorn, Silvia; Motz, Manfred; Mertens, Marc; Jilg, Wolfgang; Ulrich, Rainer G

    2012-05-01

    Hepatitis E virus (HEV) is the causative agent of an acute self-limiting hepatitis in humans. In industrialized countries, autochthonous cases are linked to zoonotic transmission from domestic pigs, wild boar and red deer. The main route of human infection presumably is consumption of contaminated meat. Farmers, slaughterers and veterinarians are expected to be risk groups as they work close to potentially infected animals. In this study, we tested four Escherichia coli-expressed segments of the capsid protein (CP) of a German wild boar-derived HEV genotype 3 strain for their diagnostic value in an indirect immunoglobulin G (IgG) ELISA. In an initial validation experiment, a carboxy-terminal CP segment spanning amino acid (aa) residues 326-608 outperformed the other segments harbouring aa residues 112-608, 326-660 and 112-335. Based on this segment, an indirect ELISA for detection of anti-HEV IgG antibodies in human sera was established and validated using a commercial line immunoassay as reference assay. A total of 563 sera from forestry workers of all forestry offices of Brandenburg, eastern Germany and 301 sera of blood donors from eastern Germany were surveyed using these assays. The commercial test revealed seroprevalence rates of 11% for blood donors and 18% for forestry workers. These rates are in line with data obtained by the in-house test (12 and 21%). Hence, the in-house test performed strikingly similar to the commercial test (sensitivity 0.9318, specificity 0.9542). An initial screening of forestry worker and blood donor sera with a corresponding CP segment of the recently discovered Norway rat-associated HEV revealed several strong positive sera exclusively in the forestry worker panel. Future investigations have to prove the performance of this novel IgG ELISA in large-scale seroepidemiological studies. In addition, the observed elevated seroprevalence in a forestry worker group has to be confirmed by studies on groups of forestry workers from other

  14. Serum antibody response to Human papillomavirus (HPV infections detected by a novel ELISA technique based on denatured recombinant HPV16 L1, L2, E4, E6 and E7 proteins

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    Mariani Luciano

    2006-11-01

    Full Text Available Abstract Background Human papillomaviruses (HPVs are the primary etiological agents of cervical cancer and are also involved in the development of other tumours (skin, head and neck. Serological survey of the HPV infections is important to better elucidate their natural history and to disclose antigen determinants useful for vaccine development. At present, the analysis of the HPV-specific antibodies has not diagnostic value for the viral infections, and new approaches are needed to correlate the antibody response to the disease outcome. The aim of this study is to develop a novel ELISA, based on five denatured recombinant HPV16 proteins, to be used for detection HPV-specific antibodies. Methods The HPV16 L1, L2, E4, E6 and E7 genes were cloned in a prokaryotic expression vector and expressed as histidine-tagged proteins. These proteins, in a denatured form, were used in ELISA as coating antigens. Human sera were collected from women with abnormal PAP smear enrolled during an ongoing multicenter HPV-PathogenISS study in Italy, assessing the HPV-related pathogenetic mechanisms of progression of cervical cancer precursor lesions. Negative human sera were collected from patients affected by other infectious agents. All the HPV-positive sera were also subjected to an avidity test to assess the binding strength in the antigen-antibody complexes. Results Most of the sera showed a positive reactivity to the denatured HPV16 proteins: 82% of the sera from HPV16 infected women and 89% of the sera from women infected by other HPV genotypes recognised at least one of the HPV16 proteins. The percentages of samples showing reactivity to L1, L2 and E7 were similar, but only a few serum samples reacted to E6 and E4. Most sera bound the antigens with medium and high avidity index, suggesting specific antigen-antibody reactions. Conclusion This novel ELISA, based on multiple denatured HPV16 antigens, is able to detect antibodies in women infected by HPV16 and it

  15. The Utility of Serum IgG4 Concentrations as a Biomarker

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    Shigeyuki Kawa

    2012-01-01

    Full Text Available IgG4-related disease is a new disease entity involving IgG4 in its clinical presentation and having 6 characteristic features: (1 systemic involvement; (2 solitary or multiple lesions showing diffuse or localized swelling, masses, nodules, and/or wall thickening on imaging; (3 high serum IgG4 concentration >135 mg/dL; (4 abundant infiltration of lymphoplasmacytes and IgG4-bearing plasma cells; (5 a positive response to corticosteroid therapy; and (6 complications of other IgG4-related diseases. To date, most IgG4-related diseases have been recognized as extrapancreatic lesions of autoimmune pancreatitis. This paper will discuss the utility of IgG4 as a biomarker of IgG4-related diseases, including in the diagnosis of autoimmune pancreatitis and its differentiation from pancreatic cancer, in the prediction of relapse, in the long-term follow-up of patients with autoimmune pancreatitis and normal or elevated IgG4 concentrations, and in patients with autoimmune pancreatitis and extrapancreatic lesions, as well as the role of IgG4 in the pathogenesis of IgG4-related disease.

  16. Testing for IgG class antibodies in celiac disease patients with selective IgA deficiency. A comparison of the diagnostic accuracy of 9 IgG anti-tissue transglutaminase, 1 IgG anti-gliadin and 1 IgG anti-deaminated gliadin peptide antibody assays.

    Science.gov (United States)

    Villalta, Danilo; Alessio, Maria Grazia; Tampoia, Marilina; Tonutti, Elio; Brusca, Ignazio; Bagnasco, Marcello; Pesce, Giampaola; Stella, Sergio; Bizzaro, Nicola

    2007-07-01

    To evaluate the diagnostic characteristics of commercially available IgG anti-tTG assays in selective IgA deficiency (SIgAD), we tested different IgG anti-tTG methods and compared the results with those obtained from two other tests: one for IgG anti-gliadin (AGA) and one for IgG to deaminated gliadin peptides (DGP). 20 CD patients with SIgAD and 113 controls (9 patients with SIgAD without CD; 54 patients with chronic liver disease; 50 healthy subjects) were tested with 9 IgG anti-tTG assays (2 of which are enriched with gliadin peptides), one IgG AGA assay and one IgG anti-DGP assay. Using optimal cutoffs as determined by ROC curves, the sensitivity of IgG anti-tTG methods ranged from 75% (1 kit) to 95% (7 kits) and the specificity from 94% (1 kit) to 100% (5 kits). Sensitivity and specificity were 40% and 87% for IgG AGA, and 80% and 98% for IgG anti-DGP, respectively. All IgG anti-tTG methods evaluated are reliable serologic assays for the diagnosis of CD in patients with SIgAD and perform better than the gliadin-based assays used in this study. The tests containing both tTG and gliadinic peptides are burdened by a lower specificity than the anti-tTG assays.

  17. Neuron-derived IgG protects neurons from complement-dependent cytotoxicity.

    Science.gov (United States)

    Zhang, Jie; Niu, Na; Li, Bingjie; McNutt, Michael A

    2013-12-01

    Passive immunity of the nervous system has traditionally been thought to be predominantly due to the blood-brain barrier. This concept must now be revisited based on the existence of neuron-derived IgG. The conventional concept is that IgG is produced solely by mature B lymphocytes, but it has now been found to be synthesized by murine and human neurons. However, the function of this endogenous IgG is poorly understood. In this study, we confirm IgG production by rat cortical neurons at the protein and mRNA levels, with 69.0 ± 5.8% of cortical neurons IgG-positive. Injury to primary-culture neurons was induced by complement leading to increases in IgG production. Blockage of neuron-derived IgG resulted in more neuronal death and early apoptosis in the presence of complement. In addition, FcγRI was found in microglia and astrocytes. Expression of FcγR I in microglia was increased by exposure to neuron-derived IgG. Release of NO from microglia triggered by complement was attenuated by neuron-derived IgG, and this attenuation could be reversed by IgG neutralization. These data demonstrate that neuron-derived IgG is protective of neurons against injury induced by complement and microglial activation. IgG appears to play an important role in maintaining the stability of the nervous system.

  18. Overview of IgG4-Related Tubulointerstitial Nephritis and Its Mimickers

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    Hyeon Joo Jeong

    2016-01-01

    Full Text Available Tubulointerstitial nephritis (TIN is the most common form of renal involvement in IgG4-related disease. It is characterized by a dominant infiltrate of IgG4-positive plasma cells in the interstitium and storiform fibrosis. Demonstration of IgG4-positive plasma cells is essential for diagnosis, but the number of IgG4-positive cells and the ratio of IgG4-positive/IgG-positive plasma cells may vary from case to case and depending on the methods of tissue sampling even in the same case. IgG4-positive plasma cells can be seen in TIN associated with systemic lupus erythematosus, Sjögren syndrome, or anti-neutrophil cytoplasmic antibody–associated vasculitis, which further add diagnostic confusion and difficulties. To have a more clear view of IgG4-TIN and to delineate differential points from other TIN with IgG4-positive plasma cell infiltrates, clinical and histological features of IgG4-TIN and its mimickers were reviewed. In the rear part, cases suggesting overlap of IgG4-TIN and its mimickers and glomerulonephritis associated with IgG4-TIN were briefly described.

  19. Metastatic Follicular Thyroid Carcinoma Secreting Thyroid Hormone and Radioiodine Avid without Stimulation: A Case Report and Literature Review

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    Syed A. Abid

    2014-01-01

    Full Text Available Introduction. This is an extremely rare case of a patient with metastatic follicular thyroid cancer who continued to produce thyroid hormone and was iodine scan positive without stimulation after thyroidectomy and radioiodine (I-131 therapy. Patient Findings. A 76-year-old Caucasian male was diagnosed with metastatic follicular thyroid carcinoma on lung nodule biopsy. Total thyroidectomy was performed and he was ablated with 160 mCi of I-131 after recombinant human thyrotropin (rhTSH stimulation. Whole body scan (WBS after treatment showed uptake in bilateral lungs, right sacrum, and pelvis. The thyroglobulin decreased from 2,063 to 965 four months after treatment but rapidly increased to 2,506 eleven months after I-131. Thyroid stimulating hormone (TSH remained suppressed and free T4 remained elevated after I-131 therapy without thyroid hormone supplementation. He was treated with an additional 209 mCi with WBS findings positive in lung and pelvis. Despite I-131, new metastatic lesions were noted in the left thyroid bed and large destructive lesion to the first cervical vertebrae four months after the second I-131 dose. Conclusions. This case is exceptional because of its rarity and also due to the dissociation between tumor differentiation and aggressiveness. The metastatic lesions continued to secrete thyroid hormone and remained radioiodine avid with rapid progression after I-131 therapy.

  20. Discovery of Radioiodinated Monomeric Anthraquinones as a Novel Class of Necrosis Avid Agents for Early Imaging of Necrotic Myocardium.

    Science.gov (United States)

    Wang, Qin; Yang, Shengwei; Jiang, Cuihua; Li, Jindian; Wang, Cong; Chen, Linwei; Jin, Qiaomei; Song, Shaoli; Feng, Yuanbo; Ni, Yicheng; Zhang, Jian; Yin, Zhiqi

    2016-02-16

    Assessment of myocardial viability is deemed necessary to aid in clinical decision making whether to recommend revascularization therapy for patients with myocardial infarction (MI). Dianthraquinones such as hypericin (Hyp) selectively accumulate in necrotic myocardium, but were unsuitable for early imaging after administration to assess myocardial viability. Since dianthraquinones can be composed by coupling two molecules of monomeric anthraquinone and the active center can be found by splitting chemical structure, we propose that monomeric anthraquinones may be effective functional groups for necrosis targetability. In this study, eight radioiodinated monomeric anthraquinones were evaluated as novel necrosis avid agents (NAAs) for imaging of necrotic myocardium. All (131)I-anthraquinones showed high affinity to necrotic tissues and (131)I-rhein emerged as the most promising compound. Infarcts were visualized on SPECT/CT images at 6 h after injection of (131)I-rhein, which was earlier than that with (131)I-Hyp. Moreover, (131)I-rhein showed satisfactory heart-to-blood, heart-to-liver and heart-to-lung ratios for obtaining images of good diagnostic quality. (131)I-rhein was a more promising "hot spot imaging" tracer for earlier visualization of necrotic myocardium than (131)I-Hyp, which supported further development of radiopharmaceuticals based on rhein for SPECT/CT ((123)I and (99m)Tc) or PET/CT imaging ((18)F and (124)I) of myocardial necrosis.

  1. Lectin-functionalized poly(glycidyl methacrylate)-block-poly(vinyldimethyl azlactone) surface supports for high avidity microbial capture

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, Ryan R [ORNL; Hinestrosa Salazar, Juan P [ORNL; Shubert, Katherine R [ORNL; Morrell, Jennifer L. [University of Tennessee, Knoxville (UTK); Pelletier, Dale A [ORNL; Messman, Jamie M [ORNL; Kilbey, II, S Michael [ORNL; Lokitz, Bradley S [ORNL; Retterer, Scott T [ORNL

    2013-01-01

    Microbial exopolysaccharides (EPS) play a critical and dynamic role in shaping the interactions between microbial community members and their local environment. The capture of targeted microbes using surface immobilized lectins that recognize specific extracellular oligosaccharide moieties offers a non-destructive method for functional characterization based on EPS content. In this report, we evaluate the use of the block co-polymer, poly(glycidyl methacrylate)-block-4,4-dimethyl-2-vinylazlactone (PGMA-b-PVDMA), as a surface support for lectin-specific microbial capture. Arrays of circular polymer supports ten micron in diameter were generated on silicon substrates to provide discrete, covalent coupling sites for Triticum vulgare and Lens culinaris lectins. These supports promoted microbe adhesion and colony formation in a lectin-specific manner. Silicon posts with similar topography containing only physisorbed lectins showed significantly less activity. These results demonstrate that micropatterned PGMA-b-PVDMA supports provide a unique platform for microbial capture and screening based on EPS content by combining high avidity lectin surfaces with three-dimensional topography.

  2. Pneumococcal polysaccharide vaccination elicits IgG anti-AB blood group antibodies in healthy individuals and patients with Type I diabetes mellitus

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    Wendelin Wolfram

    2016-11-01

    Full Text Available Hypothesis: Blood group antibodies are natural antibodies that develop early in life in response to cross-reactive environmental antigens in the absence of antigen encounter. Even later in life structural similarities in saccharide composition between environmental antigens such as bacterial polysaccharides and blood group A/B antigens could lead to changes in serum levels, IgM/IgG isotype and affinity maturation of blood group anti-A/B antibodies. We adressed the question whether immunization with pneumococcal polysaccharide (PnP vaccine (PPV Pneumovax®23 could have such an effect in patients with with type I diabetes mellitus (DM I, an autoimmune disease where an aberrant immune response to microbial antigens likely plays a role.Methods: Anti-PnP IgM and IgG responses were determined by ELISA and the Diamed-ID Micro Typing System was used to screen anti-A/B antibody titer before and after Pneumovax®23 immunization in 28 healthy individuals and 16 patients with DM I. In addition, surface plasmon resonance (SPR technology using the Biacore® device and a synthetic blood group A/B trisaccharide as the antigen was applied to investigate IgM and IgG anti-A/B antibodies and to measure antibody binding dynamics. Results: All healthy individuals and DM I patients responded with anti-PnP IgM and IgG antibody production four to six weeks after Pneumovax®23 (Pn23 immunization, while no increase in blood group anti-A/B antibody titer was observed when measured by the Diamed-ID Micro Typing System. Interestingly, isotype-specific testing by SPR-technology revealed an increase in blood group anti-A/B IgG, but not IgM, following Pn23 immunization in both patients and controls. No change in binding characteristics of blood group anti-A/B antibodies could be detected following Pn23 vaccination, supporting the assumption of an increase in IgG antibody titer with no or very little affinity maturation.Conclusion: The study provides evidence for epitope sharing

  3. Neisseria meningitidis Group A IgG1 and IgG2 Subclass Immune Response in African Children Aged 12–23 Months Following Meningococcal Vaccination

    Science.gov (United States)

    Holme, Daniel; Findlow, Helen; Sow, Samba O.; Idoko, Olubukola T.; Preziosi, Marie-Pierre; Carlone, George; Plikaytis, Brian D.; Borrow, Ray

    2015-01-01

    Background. A group A meningococcal conjugate vaccine, PsA-TT, was licensed in 2010 and was previously studied in a phase 2 clinical trial to evaluate its safety and immunogenicity in African children 12–23 months of age. Methods. Subjects received either PsA-TT; meningococcal group A, C, W, Y polysaccharide vaccine (PsACWY); or Haemophilus influenzae type b conjugate vaccine (Hib-TT). Forty weeks following primary vaccination, the 3 groups were further randomized to receive either PsA-TT, one-fifth dose of PsACWY, or Hib-TT. Group A–specific immunoglobulin G (IgG) subclass response was characterized using an enzyme-linked immunosorbent assay. Results. The predominant IgG subclass response, regardless of vaccine, was IgG1. One month following primary vaccination, the geometric mean concentrations (GMCs) of IgG1 and IgG2 in the PsA-TT group were 21.73 µg/mL and 6.27 µg/mL, whereas in the PsACWY group the mean GMCs were 2.01 µg/mL and 0.97 µg/mL, respectively (P Group A–specific IgG1 and IgG2 GMCs remained greater in the PsA-TT group than in the PsACWY group 40 weeks following primary vaccination (P vaccines. Conclusions. Vaccination of African children aged 12–24 months with either PsA-TT or PsACWY elicited a predominantly IgG1 response. The IgG1:IgG2 mean ratio decreased following successive vaccination with PsACWY, indicating a shift toward IgG2, suggestive of the T-cell–independent immune response commonly associated with polysaccharide antigens. Clinical Trials Registration. SRCTN78147026. PMID:26553689

  4. Impact of wall shear stress and ligand avidity on binding of anti-CD146-coated nanoparticles to murine tumor endothelium under flow

    Science.gov (United States)

    Ryschich, Eduard

    2015-01-01

    The endothelial phenotype of tumor blood vessels differs from the liver and forms an important base for endothelium-specific targeting by antibody-coated nanoparticles. Although differences of shear stress and ligand avidity can modulate the nanoparticle binding to endothelium, these mechanisms are still poorly studied. This study analyzed the binding of antibody-coated nanoparticles to tumor and liver endothelium under controlled flow conditions and verified this binding in tumor models in vivo. Binding of anti-CD146-coated nanoparticles, but not of antibody was significantly reduced under increased wall shear stress and the degree of nanoparticle binding correlated with the avidity of the coating. The intravascular wall shear stress favors nanoparticle binding at the site of higher avidity of endothelial epitope which additionally promotes the selectivity to tumor endothelium. After intravenous application in vivo, pegylated self-coated nanoparticles showed specific binding to tumor endothelium, whereas the nanoparticle binding to the liver endothelium was very low. This study provides a rationale that selective binding of mAb-coated nanoparticles to tumor endothelium is achieved by two factors: higher expression of endothelial epitope and higher nanoparticle shearing from liver endothelium. The combination of endothelial marker targeting and the use of shear stress-controlled nanoparticle capture can be used for selective intratumoral drug delivery. PMID:26503468

  5. Concurrent FDG Avid Nasopharyngeal Lesion and Generalized Lymphadenopathy on PET-CT Imaging Is Indicative of Lymphoma in Patients with HIV Infection

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    Yiyan Liu

    2012-01-01

    Full Text Available Patients with HIV infection often have generalized lymphadenopathy and/or other lymphoid proliferation and are at significantly increased risk for lymphoma. This study retrospectively evaluated the diagnostic value of concurrent nasopharyngeal lesion and lymphadenopathy on positron emission tomography-computed tomography (PET-CT with fluorine-18 fluorodeoxyglucose (FDG PET-CT imaging. The eligible cases were from patients with HIV infection and lymphadenopathy and referred for FDG PET-CT to evaluate lymphoma or other malignancies prior to pathological investigation. FDG PET-CT images and interpretation reports were correlated with clinical information and pathological diagnoses. Among 22 eligible patients, FDG avid nasopharyngeal lesions were incidentally noted in 7 on PET-CT imaging, and all had lymphomas diagnosed with subsequent biopsies (6 diffuse large B-cell lymphomas and 1 Hodgkin’s lymphoma. In the remaining 15 patients with adenopathy but no visible nasopharyngeal lesion or uptake on PET-CT imaging, 9 had biopsies and lymphomas were diagnosed in 4. The patients with FDG avid retroperitoneal or intra-abdominal lymphadenopathy had a greater possibility of lymphoma, compared to those with adenopathy localized only in the upper torso. Coexistent FDG avid nasopharyngeal lesion and generalized lymphadenoapthy on PET-CT imaging are indicative of a malignant lymphoma rather than benign lymphproliferative disease or nasopharyngeal carcinoma.

  6. Seroprevalence of anti-rubella and anti-measles IgG antibodies in pregnant women in Shiraz, Southern Iran: outcomes of a nationwide measles-rubella mass vaccination campaign.

    Science.gov (United States)

    Honarvar, Behnam; Moghadami, Mohsen; Moattari, Afagh; Emami, Amir; Odoomi, Neda; Bagheri Lankarani, Kamran

    2013-01-01

    Nonimmune pregnant women are at risk of developing congenital rubella syndrome and measles complications. We aimed to identify pregnant women susceptible to rubella or measles in order to determine the need for immunity screening and supplemental immunization in women of childbearing age. This seroprevalence survey was conducted by convenience sampling in obstetric hospitals affiliated with Shiraz University of Medical Sciences (southern Iran). Serum IgG levels were measured by ELISA. Mean age of the 175 pregnant women was 27.3±5.3 (range 16 to 42) years. The geometric mean concentration of anti-rubella IgG was 14.9 IU/mL (CI 95%,14.1-15.5), and that of anti-measles IgG was 13.8 IU/mL (CI 95%, 13-14.5). One hundred sixty-eight women (96%) had a protective serologic level (>11 IU/mL) of IgG against rubella, and 143 (81.7%) had a protective level against measles. Except for a significant inverse correlation that was showed by univariate analysis between anti-rubella IgG and the women's age (P = 0.01), immunity did not correlate with demographic or obstetric characteristics or medical history. There was no significant correlation between anti-rubella and anti-measles IgG levels (P = 0.25). Nearly a decade after Iran's nationwide measles-rubella vaccination campaign for the population aged 5-25 years, most pregnant women up to 34 years of age had humoral immunity against rubella. We recommend rubella immunity screening or catch-up immunization for women older than 35 years who wish to become pregnant, and measles immunity screening and appropriate vaccination for all women of childbearing age.

  7. Seroprevalence of anti-rubella and anti-measles IgG antibodies in pregnant women in Shiraz, Southern Iran: outcomes of a nationwide measles-rubella mass vaccination campaign.

    Directory of Open Access Journals (Sweden)

    Behnam Honarvar

    Full Text Available OBJECTIVE: Nonimmune pregnant women are at risk of developing congenital rubella syndrome and measles complications. We aimed to identify pregnant women susceptible to rubella or measles in order to determine the need for immunity screening and supplemental immunization in women of childbearing age. METHOD: This seroprevalence survey was conducted by convenience sampling in obstetric hospitals affiliated with Shiraz University of Medical Sciences (southern Iran. Serum IgG levels were measured by ELISA. RESULT: Mean age of the 175 pregnant women was 27.3±5.3 (range 16 to 42 years. The geometric mean concentration of anti-rubella IgG was 14.9 IU/mL (CI 95%,14.1-15.5, and that of anti-measles IgG was 13.8 IU/mL (CI 95%, 13-14.5. One hundred sixty-eight women (96% had a protective serologic level (>11 IU/mL of IgG against rubella, and 143 (81.7% had a protective level against measles. Except for a significant inverse correlation that was showed by univariate analysis between anti-rubella IgG and the women's age (P = 0.01, immunity did not correlate with demographic or obstetric characteristics or medical history. There was no significant correlation between anti-rubella and anti-measles IgG levels (P = 0.25. CONCLUSION: Nearly a decade after Iran's nationwide measles-rubella vaccination campaign for the population aged 5-25 years, most pregnant women up to 34 years of age had humoral immunity against rubella. We recommend rubella immunity screening or catch-up immunization for women older than 35 years who wish to become pregnant, and measles immunity screening and appropriate vaccination for all women of childbearing age.

  8. Methodology for determination of plasma cortisol in fish using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Cruz-Casallas, Pablo E.

    2007-01-01

    Objective. To determine plasma cortisol procedure in fish using competitive enzymelinked immunosorbent assay (ELISA). Materials and methods. Two plasma samples of juveniles rainbow trout Oncorhynchus mykiss were analized by using ELISA human kit for cortisol assay. For standard curve calibration ...

  9. IgG Antibody Responses to Recombinant gp120 Proteins, gp70V1/V2 Scaffolds, and a CyclicV2 Peptide in Thai Phase I/II Vaccine Trials Using Different Vaccine Regimens.

    Science.gov (United States)

    Karasavvas, Nicos; Karnasuta, Chitraporn; Savadsuk, Hathairat; Madnote, Sirinan; Inthawong, Dutsadee; Chantakulkij, Somsak; Rittiroongrad, Surawach; Nitayaphan, Sorachai; Pitisuttithum, Punnee; Thongcharoen, Prasert; Siriyanon, Vinai; Andrews, Charla A; Barnett, Susan W; Tartaglia, James; Sinangil, Faruk; Francis, Donald P; Robb, Merlin L; Michael, Nelson L; Ngauy, Viseth; de Souza, Mark S; Paris, Robert M; Excler, Jean-Louis; Kim, Jerome H; O'Connell, Robert J

    2015-11-01

    RV144 correlates of risk analysis showed that IgG antibodies to gp70V1V2 scaffolds inversely correlated with risk of HIV acquisition. We investigated IgG antibody responses in RV135 and RV132, two ALVAC-HIV prime-boost vaccine trials conducted in Thailand prior to RV144. Both trials used ALVAC-HIV (vCP1521) at 0, 1, 3, and 6 months and HIV-1 gp120MNgD and gp120A244gD in alum (RV135) or gp120SF2 and gp120CM235 in MF59 (RV132) at 3 and 6 months. We assessed ELISA binding antibodies to the envelope proteins (Env) 92TH023, A244gD and MNgD, cyclicV2, and gp70V1V2 CaseA2 (subtype B) and 92TH023 (subtype CRF01_AE), and Env-specific IgG1 and IgG3. Antibody responses to gp120 A244gD, MNgD, and gp70V1V2 92TH023 scaffold were significantly higher in RV135 than in RV132. Antibodies to gp70V1V2 CaseA2 were detected only in RV135 vaccine recipients and IgG1 and IgG3 antibody responses to A244gD were significantly higher in RV135. IgG binding to gp70V1V2 CaseA2 and CRF01_AE scaffolds was higher with the AIDSVAX(®)B/E boost but both trials showed similar rates of antibody decline post-vaccination. MF59 did not result in higher IgG antibody responses compared to alum with the antigens tested. However, notable differences in the structure of the recombinant proteins and dosage used for immunizations may have contributed to the magnitude and specificity of IgG induced by the two trials.

  10. Detecção de imunoglobulinas IgG, IgM e IgA anti-Toxoplasma gondii no soro, líquor e saliva de pacientes com síndrome da imunodeficiência adquirida e neurotoxoplasmose Detection of anti-Toxoplasma gondii IgG, IgM and IgA immunoglobulins in the serum, cerebrospinal fluid and saliva of patients with acquired immunodeficiency syndrome and neurotoxoplasmosis

    Directory of Open Access Journals (Sweden)

    Aercio Sebastião Borges

    2004-12-01

    Full Text Available Estudamos 55 pacientes com sindrome da imunodeficiência adquirida (SIDA e neurotoxoplasmose (grupo 1; 37 pacientes com SIDA e comprometimento neurológico por outra etiologia (grupo 2 e 18 indivíduos anti-HIV negativos com manifestações neurológicas (grupo 3, pesquisando IgG, IgA e IgM anti-Toxoplasma gondii, no soro, líquor e saliva, utilizando teste ELISA, para fins diagnósticos. O valor preditivo negativo do teste para o encontro de IgG no soro foi 100% e no líquor, 92,4%. Não houve diferença entre os três grupos quanto aos anticorpos IgA neste material. Para IgA, no líquor, o teste alcançou 72,7% de especificidade (pWe studied 55 patients with acquired immunodeficiency syndrome (AIDS and neurotoxoplasmosis (group 1, 37 patients with AIDS and neurological involvement due to another etiology (group 2 and 18 anti-HIV-negative individuals with neurological manifestations, by searching for anti-T. gondii IgG, IgA and IgM immunoglobulins in serum, cerebrospinal fluid (CSFand saliva, using ELISA. The negative predictive value of the test for IgG in serum was 100% and in CSF, 92.4%. There was no difference among the three groups studied regarding IgA in serum. For IgA, in CSF the test reached 72.7% specificity (p<0.05. In saliva, only the detection of IgG was found to be correlated with a diagnosis of neurotoxoplasmosis. We emphasize that the absence of anti-T. gondii IgG antibodies in serum and CSF strongly indicates the absence of a diagnosis of neurotoxoplasmosis and that specific IgA immunoglobulins in CSF and IgG in saliva may represent two auxiliary markers for the differential diagnosis of toxoplasmic encephalitis in AIDS.

  11. Development and Standardization of Dot - ELISA for Detection of Neospora caninum Antibodies in Cattle and Comparison with Standard Indirect ELISA and Direct Agglutination Test (DAT).

    Science.gov (United States)

    Hamidinejat, Hossein; Haji Hajikolaei, Mohamad Rahim; Ghorbanpoor, Masoud; Namavari, Mehdi; Gol, Sara Mohamad Ali

    2013-10-01

    Neospora caninum is a protozoan parasite from phylum apicomplexa and an important agent causing abortion in cattle which produce notable economic loss all around the world. Dot-Elisa was set up performing checker board procedure and then 178 sera of cattle examined with commercial indirect ELISA and direct agglutination test (DAT) were also evaluated by dot-ELISA afterwards. Kappa statistical analysis revealed that Dot-ELISA has good agreements with ELISA as well as the DAT and also, Mc Nemar's analyzing showed that this procedure has acceptable ability to discriminate positive results. Relative sensitivity and specificity of Dot-ELISA were respectively 92.63% and 89.16% and 93.4% and 90.8% in comparison with ELISA and DAT. Since the dot-ELISA is easy, inexpensive and not needed high experience to interpret the results, it is superior to ELISA and DAT when we aim to screen the cattle on the farm and slaughterhouses or when the laboratory equipment is not available.

  12. ELISA to measure neutralizing capacity of anti-C1-inhibitor antibodies in plasma of angioedema patients.

    Science.gov (United States)

    Engel, Ruchira; Rensink, Irma; Roem, Dorina; Brouwer, Mieke; Kalei, Asma; Perry, Dawn; Zeerleder, Sacha; Wouters, Diana; Hamann, Dörte

    2015-11-01

    Neutralizing autoantibodies (NAbs) against plasma serpin C1-inhibitor (C1-inh) are implicated in the rare disorder, acquired angioedema (AAE). There is insufficient understanding of the process of antibody formation and its correlation with disease progression and severity. We have developed an ELISA for detecting neutralizing capacity of anti-C1-inh positive plasma samples that can be used to study changes in NAb repertoire in patient plasma over the course of disease. The ELISA is based on the specific interaction of active C1-inh with its target protease C1s. Decrease in the amount of C1s bound to immobilized C1-inh in the presence of test samples is proportional to the neutralizing capacity of the sample. Assay specificity, intra- and inter-assay variation and assay cut-off are determined using anti-C1-inh antibodies. Assay capability is demonstrated using plasma samples from AAE patients. The assay is specific to a neutralizing anti-C1-inh antibody and shows no interference by a non-neutralizing anti-C1-inh antibody or by the plasma matrix. Intra-assay and inter-assay variations are determined as 17 and 18% respectively. Neutralizing capacity of antibody positive AAE patient plasma samples (n=16) with IgG or IgM type antibodies is readily determined. All samples show positive neutralizing capacity. We have developed a robust, specific and semi-quantitative assay to detect the neutralizing capacity of plasma samples containing anti-C1-inh antibodies. This assay can be an important tool for the study of clinical implications of anti-C1-inh NAbs. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Are Classification Criteria for IgG4-RD Now Possible? The Concept of IgG4-Related Disease and Proposal of Comprehensive Diagnostic Criteria in Japan

    Directory of Open Access Journals (Sweden)

    Kazuichi Okazaki

    2012-01-01

    Full Text Available Recent studies suggest simultaneous or metachronous lesions in multiorgans characterized by elevated serum levels of IgG4 and abundant infiltration of IgG4-positive plasma cells with various degrees of fibrosis. Two Japanese research committees for IgG4-RD, one from fibrosclerosis (Okazaki team and the other from lymph proliferation (Umehara team supported by the “Research Program for Intractable Disease” of the Ministry of Health, Labor, and Welfare of Japan, have agreed with the unified nomenclature as “IgG4-RD” and proposed the comprehensive diagnostic criteria (CDC for IgG4-RD. Validation of the CDC demonstrated satisfactory sensitivity for the practical use of general physicians and nonspecialists but low sensitivity in the organs to be difficult in taking biopsy specimens such as type1 autoimmune pancreatitis (IgG4-related AIP, compared with IgG4-related sialadenitis/dacryoadenitis (Mikulicz's disease and IgG4-related kidney disease. Although the diagnostic criteria covering all IgG4-RD are hard to be established, combination with the CDC and organ-specific diagnostic criteria should improve sensitivity.

  14. IMPROVEMENT OF THE QUALITY CONTROL OF ELISA TESTING FOR THE LABORATORY CONFIRMATION OF MEASLES AND RUBELLA INFECTIONS AT THE STAGE OF THE MEASLES/ RUBELLA ELIMINATION PROGRAM

    Directory of Open Access Journals (Sweden)

    T. A. Mamaeva

    2017-01-01

    Full Text Available To estimate ELISA serological studies results of IgM and IgG specific Measles and Rubella Viruses (MRV antibodies detection the “in-house” laboratory controls (ILC including the specific markers of MRV infections were for the first time commercially prepared by the Vector Best PLC (Russia: “Measles-IgM, ser.1”, “Measles-IgM, ser.2”, “Rubella-IgM”, Measles-IgG” and “Rubella-IgG”. This task was realized under the special Executive Order of the Government of Russia N 523-r, 2014, April, 4. According to passport characteristics ILC samples are the lyophilized human sera, inactivated by heating (1 hour at 56°C and stabilized by the mixture of sucrose (5% and ProClin-3000 as the conservation agent. Samples are free of HBs Ag, anti-HVC, T.Pallidum, HIV-1/2, HIV-1Ag р24.The aim of the study was to evaluate the possibility of using the ILC for detection of the MRV IgM and IgG antibodies by ELISA with commercial ELISA kits used in Russia and CIS countries. In the process of detecting the specific activity of “Measles-IgM, ser.1”, “Measles-IgM, ser.2” and “Rubella-IgM” by ELISA kits of different formats (Vector Best, EcoLab and Siemens Companies the statistically different results were received (p < 0.05. The optical density (OD values of IgM in the “Measles-IgM, ser.1” and “Measles-IgM, ser.2” ILC, obtained by ELISA “VectoMeasles IgM” (Vector Best were significantly higher than those obtained by ELISA IgEnzygnost®Anti-MeaslesVirus/IgМ. These values consisted for the ser. 1–1.33±0.02 о.u. vs. 0.18±0.01 о.u. (р < 0.05 and for the ser. 2–2.83±0.03 о.u. vs. 0.7±0.02 о.е. (р < 0.05 in the Vector Best and Siemens ELISA kits correspondently. In the “Rubella-IgM” ILC the OD values of the specific IgM by the “ELISA-Rubella IgM” EcoLab were also higher than those obtained by IgEnzygnost®Anti-RubellaVirus/IgМ ELISA kit. These values consisted 2.92±0.04 о.u. vs. 0.88±0.03

  15. The ELisA Facility - RESTful API and Client Libraries

    CERN Document Server

    Corso-Radu, A; The ATLAS collaboration; Murillo Garcia, R

    2013-01-01

    The ATLAS experiment at the LHC (CERN) comprises a large and geographically distributed community of over three thousand scientists from all over the world. Data acquisition is supervised by a shift crew of about 10 people running the experiment 24/7. Information concerning the experiment operation, configuration and behavior has to be reported, gathered and shared with the whole community. To provide such functionality, a logbook facility tool, known as ELisA, has been developed to offer an user-friendly web interface to browse activity logs and an effective way for shifters and experts to report on system operations.\

  16. Rubella virus detection by ELISA method in exposed radiation workers

    International Nuclear Information System (INIS)

    Wu Jianmei; Zhu Bo; Zhu Youming; Shao Jinhui; Wu Weiping; Han Jinxiang

    2005-01-01

    Objective: A rapid diagnosis method was developed to detect Rubella virus infection in radiation workers. Methods: Modified ELISA method was used to detect the level of lgG and lgM antibodies in 514 in Jinan district. Results: 90.47% of 514 cases was shown to be resistant against Rubella virus; 6.42% were sensitive type; 0.78% belonged to be reinfected. Conclusion: Detection of Rubella virus in exposed radiation workers was imperative, and vaccine against Rubella virus was also needed to eliminate the infection risk. (authors)

  17. Evaluation Of Antibody Elisa, Coproscopy And Serum Enzyme ...

    African Journals Online (AJOL)

    Le titrage avec immunoadsorbant lié à une enzyme (ELISA), la sédimentation fécale et les tests de l'action de l'enzyme du sérum ont été faits sur des échantillons de fèces et de sérum recueillis de 134 bovins (55 positifs et 79 négatifs pour les lésions dues à la douve du foie) lors de l'inspection de viande en Ethiopie.

  18. Estimating False-Recent Classification for the Limiting-Antigen Avidity EIA and BED-Capture Enzyme Immunoassay in Vietnam: Implications for HIV-1 Incidence Estimates.

    Science.gov (United States)

    Shah, Neha S; Duong, Yen T; Le, Linh-Vi; Tuan, Nguyen Anh; Parekh, Bharat S; Ha, Hoang Thi Thanh; Pham, Quang Duy; Cuc, Cao Thi Thu; Dobbs, Trudy; Tram, Tran Hong; Lien, Truong Thi Xuan; Wagar, Nick; Yang, Chunfu; Martin, Amy; Wolfe, Mitchell; Hien, Nguyen Tran; Kim, Andrea A

    2017-06-01

    Laboratory tests that can distinguish recent from long-term HIV infection are used to estimate HIV incidence in a population, but can potentially misclassify a proportion of long-term HIV infections as recent. Correct application of an assay requires determination of the proportion false recents (PFRs) as part of the assay characterization and for calculating HIV incidence in a local population using a HIV incidence assay. From April 2009 to December 2010, blood specimens were collected from HIV-infected individuals attending nine outpatient clinics (OPCs) in Vietnam (four from northern and five from southern Vietnam). Participants were living with HIV for ≥1 year and reported no antiretroviral (ARV) drug treatment. Basic demographic data and clinical information were collected. Specimens were tested with the BED capture enzyme immunoassay (BED-CEIA) and the Limiting-antigen (LAg)-Avidity EIA. PFR was estimated by dividing the number of specimens classified as recent by the total number of specimens; 95% confidence intervals (CI) were calculated. Specimens that tested recent had viral load testing performed. Among 1,813 specimens (north, n = 942 and south, n = 871), the LAg-Avidity EIA PFR was 1.7% (CI: 1.2-2.4) and differed by region [north 2.7% (CI: 1.8-3.9) versus south 0.7% (CI: 0.3-1.5); p = .002]. The BED-CEIA PFR was 2.3% (CI: 1.7-3.0) and varied by region [north 3.4% (CI: 2.4-4.7) versus south 1.0% (CI: 0.5-1.2), p < .001]. Excluding specimens with an undetectable VL, the LAg-Avidity EIA PFR was 1.2% (CI: 0.8-1.9) and the BED-CEIA PFR was 1.7% (CI: 1.2-2.4). The LAg-Avidity EIA PFR was lower than the BED-CEIA PFR. After excluding specimens with an undetectable VL, the PFR for both assays was similar. A low PFR should facilitate the implementation of the LAg-Avidity EIA for cross-sectional incidence estimates in Vietnam.

  19. Presence of Mycoplasma fermentans in the bloodstream of Mexican patients with rheumatoid arthritis and IgM and IgG antibodies against whole microorganism

    Directory of Open Access Journals (Sweden)

    Salinas Salvador

    2009-08-01

    Full Text Available Abstract Background Increasing evidence incriminates bacteria, especially Mycoplasma fermentans, as possible arthritogenic agents in humans. The purpose of this study was to investigate M. fermentans in the bloodstream of patients with rheumatoid arthritis. Methods Two hundred and nineteen blood samples from patients with rheumatoid arthritis, systemic lupus erythematosus, antiphospholipid syndrome, and healthy individuals were screened by bacterial culture and direct PCR in order to detect mycoplasmas; IgM and IgG against M. fermentans PG18 were also detected by ELISA and Immunoblotting assays in patients with rheumatoid arthritis and healthy individuals. Results Blood samples from patients with antiphospholipid syndrome and healthy individuals were negative for mycoplasma by culture or direct PCR. In blood samples from patients with systemic lupus erythematosus were detected by direct PCR M. fermentans in 2/50 (2%, M. hominis in 2/50 (2% and U. urealyticum in 1/50 (0.5%. In patients with RA M. fermentans was detected by culture in 13/87 blood samples and in 13/87 by direct PCR, however, there was only concordance between culture and direct PCR in six samples, so M. fermentans was detected in 20/87(23% of the blood samples from patients with RA by either culture or PCR. Antibody-specific ELISA assay to M. fermentans PG18 was done, IgM was detected in sera from 40/87 patients with RA and in sera of 7/67 control individuals, IgG was detected in sera from 48/87 RA patients and in sera from 7/67 healthy individuals. Antibody-specific immunoblotting to M. fermentans PG18 showed IgM in sera from 35/87 patients with RA and in sera from 4/67 healthy individuals, IgG was detected in sera from 34/87 patients and in sera from 5/67 healthy individuals. Conclusion Our findings show that only M. fermentans produce bacteremia in a high percentage of patients with RA. This finding is similar to those reported in the literature. IgM and IgG against M

  20. Evaluation of an ELISA using recombinant Ssλ20ΔB3 antigen for the serological diagnosis of Sarcoptes scabiei infestation in domestic and wild rabbits.

    Science.gov (United States)

    Casais, Rosa; Millán, Javier; Rosell, Joan Maria; Dalton, Kevin P; Prieto, José Miguel

    2015-12-15

    An ELISA, based on the Sarcoptes scabiei Ssλ20ΔB3 inmunodominant antigen, was evaluated for the detection of antibodies to S. scabiei in experimentally infested (n=10), farm (n=109), and wild (n=78) rabbit sera. The S. scabiei antigen Ssλ20ΔB3, a major structural protein present over the entire mite's body, was produced as a recombinant protein in Escherichia coli and purified for its use in the ELISA. The resulting ELISA showed, in experimentally infested domestic rabbits, detectable specific antibody responses (IgG) above the cut off level from week three post-infestation indicating that the assay is able to detect positive rabbits very early during the course of the infestation. The ELISA was validated on a panel of 109 domestic breeding rabbit sera collected from 26 Spanish farms, of which 41 were obtained from rabbits with skin lesions compatible with sarcoptic mange, 26 with skin lesions compatible with psoroptic mange, and 42 from unexposed individuals from mange-free farms. The ELISA in this group was characterized by 95% sensitivity, 97% specificity, and a high degree of repeatability. In the psoroptic mange compatible lesions group, included in the study as control group for cross-reactivity with the closely related mite Psoroptes cuniculi, cross-reacting antibodies to Ssλ20ΔB3 S. scabiei antigen were detected in 42.30% of the rabbit sera. However, mean% OD values of the sarcoptic-mange group (55.61 ± 39.20%) were significantly higher (psarcoptic mange compatible lesions and presence of S. scabiei mites were seropositive) and the specificity was 90% (67 out of 74 wild rabbits without detectable mange lesions were seronegative). Although, the total number of tested samples from experimentally infested, farm and wild rabbits was limited, our study showed that the ELISA is able to differentiate between infested and non-infested animals in all tested groups with very high sensitivity and specificity indicating that recombinant Ssλ20ΔB3 is a reliable

  1. Simultaneous immunolocalization of desmoglein 3 and IgG4 in oral pemphigus vulgaris: IgG4 predominant autoantibodies in its pathogenesis.

    Science.gov (United States)

    Abé, Tatsuya; Maruyama, Satoshi; Babkair, Hamzah; Yamazaki, Manabu; Cheng, Jun; Saku, Takashi

    2015-11-01

    Oral pemphigus vulgaris (PV), an autoimmune blistering disease, is mainly mediated by autoantibodies against desmoglein (Dsg) 3. However, no attention has been paid to IgG subclasses of the autoantibodies against Dsg3 in the diagnostic procedure for PV. Thus, our aim in this study was to investigate whether Dsg3 and any of IgG subclasses are immunohistochemically colocalized in tissue sections of PV oral mucosa. Serial sections cut from formalin-fixed paraffin blocks of biopsy specimens of 9 PV cases and those of normal buccal mucosa surgically removed for fibro-epithelial polyps were comparatively examined for immunohistochemical localizations for Dsg3, IgG4, and IgG. Dsg3 was demonstrated in a dot-like pattern on the cell border and in the cytoplasm of the whole epithelial layer in both normal and PV specimens, while its staining was irregular among floating epithelial sheets of PV. IgG4 was also demonstrated in a punctuated fashion on the cell border among floating epithelial sheets, which was nearly identical to the immunohistochemical profile of Dsg3. In addition to being detected in the epithelial part, IgG4 signals were prominently localized in plasma cells scattered in the granulation tissue, where ratios of IgG4-positive (+) plasma cells to IgG+ cells were extraordinarily higher (mean 28%) than those in normal mucosa. These findings confirmed for the first time that autoantibodies against Dsg3 are mainly composed of IgG4 in oral PV and that the combined immunohistochemistry for Dsg3 and IgG4 can be a valuable aid in confirming a histopathological diagnosis of PV. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  2. IgG4-related mastitis, a rare disease, can radiologically and histologically mimic malignancy.

    Science.gov (United States)

    Yamada, Rin; Horiguchi, Shin-ichiro; Yamashita, Toshinari; Kamisawa, Terumi

    2016-03-23

    IgG4-related disease (IgG4-RD) is characterised by high serum concentrations of IgG4, dense lymphoplasmacytic infiltrates, storiform fibrosis and increased IgG4-positive plasma cells in tissues. This systemic disease occurs in various organs metachronously, but IgG4-related mastitis appears extremely rare. We report a case of IgG4-related mastitis, radiologically considered to represent breast cancer mainly composed of intraductal component and requiring histological differentiation from mucosa-associated lymphoid tissue (MALT) lymphoma. The breast mass disappeared with steroid therapy. When patients have a breast mass, regardless of the presence or absence of IgG4-RD, IgG4-related mastitis should be considered in addition to breast cancer. If histological findings show dense lymphoplasmacytic infiltrates, IgG4-related mastitis should be suspected in addition to malignant lymphoma, and lack of monoclonality should be confirmed. To avoid unnecessary surgery or chemotherapy, knowledge and accurate diagnosis of the entity of IgG4-related mastitis is necessary. 2016 BMJ Publishing Group Ltd.

  3. Severe IgG4-Related Disease in a Young Child: A Diagnosis Challenge

    Directory of Open Access Journals (Sweden)

    Susana Corujeira

    2015-01-01

    Full Text Available Immunoglobulin G4-related disease (IgG4-RD is an increasingly recognized syndrome that can appear with multiple organ involvement, typically with tumor-like swelling, lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells, and elevated serum IgG4 concentrations. We report the case of a 22-month-old female child with failure to thrive and recurrent respiratory tract infections since 8 months of age. Physical examination was normal except for pulmonary auscultation with bilateral crackles and wheezes. Laboratory tests revealed elevated erythrocyte sedimentation rate, and elevated serum IgG and IgG4 with polyclonal hypergammaglobulinemia. Thoracic CT and MRI showed multiple mediastinal lymphadenopathies and a nodular posterior mediastinal mass in right paratracheal location with bronchial compression. Initial fine needle aspiration biopsy was compatible with reactive lymphadenopathy but after clinical worsening a thoracoscopic partial resection of the mass was performed and tissue biopsy revealed lymphoplasmacytic infiltrate and increased number of IgG4-positive plasma cells and a ratio of IgG4/IgG positive cells above 40%. Glucocorticoids therapy was started with symptomatic improvement, reduction in the size of the mass, and decrease of serum IgG4 levels after 6 weeks. There are very few reports of IgG4-RD in children. Long-term follow-up is necessary to monitor relapses and additional organ involvement.

  4. ELISA, a demonstrator environment for information systems architecture design

    Science.gov (United States)

    Panem, Chantal

    1994-01-01

    This paper describes an approach of reusability of software engineering technology in the area of ground space system design. System engineers have lots of needs similar to software developers: sharing of a common data base, capitalization of knowledge, definition of a common design process, communication between different technical domains. Moreover system designers need to simulate dynamically their system as early as possible. Software development environments, methods and tools now become operational and widely used. Their architecture is based on a unique object base, a set of common management services and they host a family of tools for each life cycle activity. In late '92, CNES decided to develop a demonstrative software environment supporting some system activities. The design of ground space data processing systems was chosen as the application domain. ELISA (Integrated Software Environment for Architectures Specification) was specified as a 'demonstrator', i.e. a sufficient basis for demonstrations, evaluation and future operational enhancements. A process with three phases was implemented: system requirements definition, design of system architectures models, and selection of physical architectures. Each phase is composed of several activities that can be performed in parallel, with the provision of Commercial Off the Shelves Tools. ELISA has been delivered to CNES in January 94, currently used for demonstrations and evaluations on real projects (e.g. SPOT4 Satellite Control Center). It is on the way of new evolutions.

  5. Detecção pelo teste imunoenzimático ELISA de anticorpos IgM anti-Cysticercus cellulosae no líquido cefalorraqueano na neurocisticercose

    Directory of Open Access Journals (Sweden)

    Julia M. Costa

    1985-03-01

    Full Text Available Demonstrou-se a presença de anticorpos da classe IgM anti-C. cellulosaeem amostras de LCR de pacientes com neurocisticercose, pelo teste imunoenzimático ELISA. Este foi realizado em placas plásticas sensibilizadas com uma fração glicoproteica de cisticercos. Das 41 amostras de LCR estudadas, 26 pertenciam a pacientes com neurocisticercose, 5 a pacientes com neurossífilis e 10 a pessoas aparentemente normais. Nas 5 amostras de LCR de pacientes com neurossífilis e nas 10 de pessoas aparentemente normais foi negativo o teste ELISA-IgM. Dos 26 LCR de pacientes com neurocisticercose 12 (46,2% apresentaram anticorpos IgM anti-C. cellulosaecom títulos que variaram de 4 a 32. Essas 12 amostras de LCR quando submetidas ao tratamento com 2-mercaptoetanol tornaram-se negativas no teste ELISA-IgM. Compararam-se os níveis de anticorpos IgM e IgG anti-C. cellulosaedetectados pelos respectivos testes imunoenzimáticos para todas as amostras estudadas e observou-se que, dos 12 LCR de pacientes com neurocisticercose reagentes pelos dois testes, dois apresentaram níveis de IgM mais elevados que de IgG. Paralelamente compararam-se os resultados dos testes ELISA com as reações de fixação do complemento, imunofluorescência e hemaglutinação para neurocisticercose.

  6. Normal physiologic and Benign foci with F-18 FDG avidity on PET/CT in patients with breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Park, Soon Ah; Lee, Kwang Man; Choi, Un Jong; Kim, Hun Soo; Kim, Hye Won; Song, Jeong Hoon [College of Medicine, Wonkwnag University, Iksan (Korea, Republic of)

    2010-12-15

    The aim of this study was to evaluate the physiologic and benign F-18 fluorodeoxyglucose (FDG) avid foci in patients with breast cancer. On 309 F-18 FDG PET/CT scans of 241 women with breast cancer, the hypermetabolic lesions compared with the surrounding normal region were evaluated retrospectively. Available reports of other relevant radiological imaging medical records, and follow-up PET/CT were reviewed for explanations of the abnormal uptake. Among the 70 physiologic foci, muscular uptake of the lower neck following the surgical and/or radiation therapy of ipsilateral breast (29%), hypermetabolic ovaries (16%) and uterine (10%) uptake during the ovulatory and menstrual phases during the normal menstrual cycle were identified, and also hypermetabolic brown fat in cold-induced thermogenesis (7%), non-specific bowel uptake (35%) were observed. Among the 147 benign lesions, sequelae of the chest wall and breasts following surgical and/or radiation therapy, were often observed (27%). Hypermetabolic thyroid glands were noted as adenomas and chronic thyroiditis (18%). Reactive hyperplasia of cervical or mediastinal lymph nodes (32%), degenerative osteoarthritis and healed fractures (15%), hypermetabolic benign lung lesions (6%) were observed. Altered physiologic and benign F-18 FDG uptake in the cervical muscle and chest wall following ipsilateral breast surgery or radiotherapy were common, and also normal physiologic uptake in ovary and uterus, brown fat, thyroid were considered as predominant findings in women patients with breast cancer. Knowledge of these findings might aid in the interpretation of FDG PET/CT in patients with breast cancer

  7. Evaluation of two Taenia solium cysticercal antigenic preparations (vesicular fluid and a glycoprotein fraction with affinity for lentil lectin for the immunodiagnosis of neurocysticercosis by enzyme-linked immunosorbent assay (ELISA

    Directory of Open Access Journals (Sweden)

    Lisandra Akemi Suzuki

    2011-06-01

    Full Text Available OBJECTIVE: To evaluate the performance of two antigenic preparations (vesicular fluid - VF and a glycoprotein fraction, LLa-Gp fraction, purified from a whole parasite extract by lentil lectin affinity chromatography from Taenia solium cysticerci for the immunodiagnosis of neurocysticercosis. METHOD: Fifty-six cerebrospinal fluid (CSF samples (22 from patients with neurocysticercosis and 34 from patients with other neurological disorders and 57 serum samples (22 from patients with neurocysticercosis, 18 from patients with other infections and 17 from presumably healthy persons were assayed for anticysticercal IgG antibodies with an enzyme-linked immunosorbent assay (ELISA. RESULTS: The VF ELISA showed 100% sensitivity and specificity in CSF and serum samples, whereas the sensitivity and specificity of the LLa-Gp ELISA were, respectively, 90.9% and 97.1%, with the CSF samples and 95.5% and 100% with serum samples. There was no significant difference in the sensitivity and specificity of the two antigenic preparations used to screen CSF and serum samples. CONCLUSION: Considering the complexity and high cost of obtaining the LLa-Gp fraction, VF could be more suitable for screening specific antibodies by ELISA in CSF and serum samples from patients with neurocysticercosis.

  8. Development of a novel affinity chromatography resin for platform purification of bispecific antibodies with modified protein a binding avidity.

    Science.gov (United States)

    Tustian, Andrew D; Laurin, Linus; Ihre, Henrik; Tran, Travis; Stairs, Robert; Bak, Hanne

    2018-02-21

    There is strong interest in the production of bispecific monoclonal antibodies that can simultaneously bind two distinct targets or epitopes to achieve novel mechanisms of action and efficacy. Regeneron's bispecific technology, based upon a standard IgG, consists of a heterodimer of two different heavy chains, and a common light chain. Co-expression of two heavy chains leads to the formation of two parental IgG impurities, the removal of which is facilitated by a dipeptide substitution in the Fc portion of one of the heavy chains that ablates Fc Protein A binding. Therefore the affinity capture (Protein A) step of the purification process must perform both bulk capture and high resolution of these mAb impurities, a task current commercially available resins are not designed for. Resolution can be further impaired by the ability of Protein A to bind some antibodies in the variable region of the heavy chain (V H ). This paper details development of a novel Protein A resin. This resin combines an alkali stable ligand with a base matrix exhibiting excellent mass transfer properties to allow high capacity single step capture and resolution of bispecific antibodies with high yields. The developed resin, named MabSelect SuRe™ pcc, is implemented in GMP production processes for several bispecific antibodies. This article is protected by copyright. All rights reserved. © 2018 American Institute of Chemical Engineers.

  9. Validation of an excretory/secretory antigen based-ELISA for the diagnosis of Opisthorchis felineus infection in humans from low trematode endemic areas.

    Science.gov (United States)

    Gómez-Morales, Maria Angeles; Ludovisi, Alessandra; Amati, Marco; Pozio, Edoardo

    2013-01-01

    Since opisthorchiasis does not show pathognomonic signs or symptoms, physicians can have serious problems to make a differential diagnosis of this infection in non endemic areas, in particular when there is a simultaneous occurrence with other seasonal infections. Moreover, symptomatic infections due to O. felineus can last a few weeks and then the signs and symptoms disappear, but the worms survive in the bile ducts for years causing hepatobiliary diseases including hepatomegaly, cholangitis, fibrosis of the periportal system, cholecystitis, and gallstones. Consequently, an early diagnosis prevents chronicity and loss of working days. The detection of specific antibodies has been considered as a complementary tool to the fecal examination to establish the definitive diagnosis of this infection and for the follow up. Therefore the aim of this work was the development and validation of an enzyme-linked immunosorbent assay (ELISA) using excretory/secretory antigens (ESA) from O. felineus adult worms to detect anti-Opisthorchis IgG in human sera. A total of 370 human sera were tested: 144 sera from persons with a confirmed diagnosis of opisthorchiasis, 110 sera from healthy Italian people, and 116 sera from people with other parasitic or non-parasitic infections. Results were analyzed by receiver-operator characteristic (ROC) curve analysis. The accuracy of the test, calculated by the area under curve (AUC), yielded a 0.999 value, indicating the high performance of the test. The sensitivity was 100% (95% CI: 97.40% to 100%) and no false-negative sera were detected; the specificity was 99.09% (95% CI: 95.02% to 99.83%). The validated ELISA shows a good performance in terms of sensitivity, repeatability and reproducibility, and it is suitable to detect anti-Opisthorchis IgG in human sera for diagnostic purposes and for the follow up to assess the efficacy of drug treatment.

  10. Validation of an excretory/secretory antigen based-ELISA for the diagnosis of Opisthorchis felineus infection in humans from low trematode endemic areas.

    Directory of Open Access Journals (Sweden)

    Maria Angeles Gómez-Morales

    Full Text Available Since opisthorchiasis does not show pathognomonic signs or symptoms, physicians can have serious problems to make a differential diagnosis of this infection in non endemic areas, in particular when there is a simultaneous occurrence with other seasonal infections. Moreover, symptomatic infections due to O. felineus can last a few weeks and then the signs and symptoms disappear, but the worms survive in the bile ducts for years causing hepatobiliary diseases including hepatomegaly, cholangitis, fibrosis of the periportal system, cholecystitis, and gallstones. Consequently, an early diagnosis prevents chronicity and loss of working days. The detection of specific antibodies has been considered as a complementary tool to the fecal examination to establish the definitive diagnosis of this infection and for the follow up. Therefore the aim of this work was the development and validation of an enzyme-linked immunosorbent assay (ELISA using excretory/secretory antigens (ESA from O. felineus adult worms to detect anti-Opisthorchis IgG in human sera. A total of 370 human sera were tested: 144 sera from persons with a confirmed diagnosis of opisthorchiasis, 110 sera from healthy Italian people, and 116 sera from people with other parasitic or non-parasitic infections. Results were analyzed by receiver-operator characteristic (ROC curve analysis. The accuracy of the test, calculated by the area under curve (AUC, yielded a 0.999 value, indicating the high performance of the test. The sensitivity was 100% (95% CI: 97.40% to 100% and no false-negative sera were detected; the specificity was 99.09% (95% CI: 95.02% to 99.83%. The validated ELISA shows a good performance in terms of sensitivity, repeatability and reproducibility, and it is suitable to detect anti-Opisthorchis IgG in human sera for diagnostic purposes and for the follow up to assess the efficacy of drug treatment.

  11. High seroprevalence of Mycoplasma pneumoniae IgM in acute Q fever by enzyme-linked immunosorbent assay (ELISA.

    Directory of Open Access Journals (Sweden)

    Chung-Hsu Lai

    Full Text Available Q fever is serologically cross-reactive with other intracellular microorganisms. However, studies of the serological status of Mycoplasma pneumoniae and Chlamydophila pneumoniae during Q fever are rare. We conducted a retrospective serological study of M. pneumoniae and C. pneumoniae by enzyme-linked immunosorbent assay (ELISA, a method widely used in clinical practice, in 102 cases of acute Q fever, 39 cases of scrub typhus, and 14 cases of murine typhus. The seropositive (57.8%, 7.7%, and 0%, p<0.001 and seroconversion rates (50.6%, 8.8%, and 0%, p<0.001 of M. pneumoniae IgM, but not M. pneumoniae IgG and C. pneumoniae IgG/IgM, in acute Q fever were significantly higher than in scrub typhus and murine typhus. Another ELISA kit also revealed a high seropositivity (49.5% and seroconversion rate (33.3% of M. pneumoniae IgM in acute Q fever. The temporal and age distributions of patients with positive M. pneumoniae IgM were not typical of M. pneumoniae pneumonia. Comparing acute Q fever patients who were positive for M. pneumoniae IgM (59 cases with those who were negative (43 cases, the demographic characteristics and underlying diseases were not different. In addition, the clinical manifestations associated with atypical pneumonia, including headache (71.2% vs. 81.4%, p=0.255, sore throat (8.5% vs. 16.3%, p=0.351, cough (35.6% vs. 23.3%, p=0.199, and chest x-ray suggesting pneumonia (19.3% vs. 9.5%, p=0.258, were unchanged between the two groups. Clinicians should be aware of the high seroprevalence of M. pneumoniae IgM in acute Q fever, particularly with ELISA kits, which can lead to misdiagnosis, overestimations of the prevalence of M. pneumoniae pneumonia, and underestimations of the true prevalence of Q fever pneumonia.

  12. Study on the seroprevalence of toxoplasmosis in sheep and goats of Tabriz area using the ELISA method

    Directory of Open Access Journals (Sweden)

    H Hashemzadeh Farhang

    2010-05-01

    Full Text Available Toxoplasmosis is one of the most important zoonotic diseases and causes abortion in sheep and goats. In order to detect the slaughterhouse infection rate of toxoplasmosis in sheep and goats of Tabriz area, 186 blood sera samples were prepared and examined for IgG antibodies against toxoplasmosis using the ELISA method. Serological results indicated that the infection rate of Toxoplasma gondii was 18.3% with 13.45% of the infection rate related to sheep and 4.85% related to goats. The results indicate significant difference in the infection rate between sheep and goats of the region which could be related to the higher susceptibility of sheep to toxoplasmosis in comparison with goats. Based on the results of the study the infection rate in females was higher than males with the infection rate in male sheep being 11.5% while that of the females was 25.8% indicating signification difference in infection rates between males and females. The infection rate in male and female goats was 9.1% and 10.8% respectively and no significant difference in infection rate was observed between male and female goats.

  13. Immunoglobulin G (IgG) Expression in Human Umbilical Cord Endothelial Cells

    Science.gov (United States)

    Zhao, Yingying; Liu, Yuxuan; Chen, Zhengshan; Korteweg, Christine; Gu, Jiang

    2011-01-01

    Traditional views hold that immunoglobulin G (IgG) in the human umbilical cord is internalized by human umbilical endothelial cells for passive immunity. In this study, the protein and mRNA transcripts of IgG were found in the cytoplasm of human umbilical endothelial cells by immunohistochemistry, in situ hybridization, and reverse transcription PCR (RT-PCR). The essential enzymes for IgG synthesis and assembling, RAG1 (recombination activating gene 1), RAG2, and variable (V), diversity (D), and joining (J) segments for recombination of IgG, were also found in these cells by RT-PCR and real-time PCR. These results indicate that umbilical endothelial cells are capable of synthesizing IgG with properties similar to those of immune cells and that they may play additional roles besides lining the vessels and transporting IgG. PMID:21430258

  14. [Severe asthmatic crisis during general anesthesia in a patient with IgG4 related disease].

    Science.gov (United States)

    Moriya, Machika; Oda, Shinya; Nakane, Masaki; Kawamae, Kaneyuki

    2014-04-01

    We experienced severe asthmatic crisis during general anesthesia in a 45-year-old man with IgG4-related disease, COPD and athma undergoing removal of submandibular gland. The ventilatiory failure was caused by the stimulation of the operation, sputum, and neostigmine. His serum IgG4 level was extremely high. IgG4 related disease is a recently emerging entity characterized by a diffuse or mass forming inflammatory reaction rich in IgG4-positive plasma cells associated with fibrosclerosis and obliterative phlebitis. It is associated with an elevated serum level of IgG4 and an allergic disease. We must be careful in perioperative management of the patients with IgG4-related disease because general anesthesia can induce asthmatic crisis.

  15. SIMPLIFIED DIAGNOSIS OF MALARIA INFECTION: GFM/PCR/ELISA A SIMPLIFIED NUCLEIC ACID AMPLIFICATION TECHNIQUE BY PCR/ELISA

    Directory of Open Access Journals (Sweden)

    Ricardo Luiz Dantas MACHADO

    1998-09-01

    Full Text Available We report an adaptation of a technique for the blood sample collection (GFM as well as for the extraction and amplification of Plasmodium DNA for the diagnosis of malaria infection by the PCR/ELISA. The method of blood sample collection requires less expertise and saves both time and money, thus reducing the cost by more than half. The material is also suitable for genetic analysis in either fresh or stored specimens prepared by this method.Relatamos a adaptação de uma técnica para coleta de amostras (MFV e outra para extração, amplificação de DNA de parasitas da malária para diagnóstico por PCR/ELISA. O método de coleta de amostras requer menos habilidade e economisa tempo e dinheiro, assim reduzindo a mais da metade o custo. O material é também adequado para análise genética em especimens frescos ou estocados, preparados por este método.

  16. A three-layer immunoradiometric assay for determination of IgG subclass antibodies in Human Sera (''IgG subclass RAST'')

    International Nuclear Information System (INIS)

    Djurup, R.; Soendergaard, I.; Weeke, B.; University of Copenhagen, Denmark); Magnusson, C.G.M.

    1984-01-01

    We report the development of a three-layer immunoradiometric assay (TIRA) for measurement of IgG antibodies of all four subclasses in human sera. The first layer consists of diluted human serum, the second layer is monoclonal mouse antibodies to human IgG subclasses, and the third layer is 125 I-labelled rabbit anti-mouse IgG. Monoclonal anti-IgGI, anti-IgG3 and anti-IgG4 reacted only with their complementary IgG subclass, whereas the anti-IgG2 showed slight cross-reactivity to immunoglobins of other subclasses and classes and to light chain proteins. The observed cross-reactivity was found to be without importance, when the TIRA was applied to measurement of IgG subclass antibodies. Equipotency was established by use of appropriate dilutions of the monoclonal antibodies, and the assay was calibrated by use of human reference serum. The TIRA therefore permits reliable inter-individual and intra-individual comparisons of the IgG antibody response in all four subclasses. Non-specific binding obtained with pooled normal human serum was below 0.33%. Inter-assay coefficient of variation was between 18 and 27%. The TIRA was applied to measurement of IgG subclass antibodies to timothy grass pollen in sera from grass pollen allergies undergoing immunotherapy. (author)

  17. Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis

    DEFF Research Database (Denmark)

    Johnsen, C R; Abrahamsen, L; Stahl Skov, P

    1991-01-01

    of indoor aeroallergens, cat, dog, and Derm. pter. allergen extracts were selected for the experiments. To evaluate unspecific interference, these allergens were compared mutually and with Cladosporium herbarum. Allergen extracts in varying dilutions were mixed with crushed glass fibre filter materials......, eluted, recovered by centrifugation, and allergen concentration quantified by the assays. Equal sensitivity was found for both IgE- and IgG4-RI assaying cat allergen (in the range 5-50 SQ-U/ml) and dog allergen (in the range 10(2)-10(3) SQ-U/ml). The IgG4-RI assaying Derm. pter. was more sensitive (50 SQ......-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat...

  18. The Relationship of Gamma Immunoglobin (IgG) Density and Apgar ...

    African Journals Online (AJOL)

    The transfer of maternal IgG provides the neonate with humoral immunity during early life. The population of transferred IgG or IgG density (IgGρ) was estimated to find out if it has any relevance to the condition of an infant 1-5 minutes after birth or APGAR score which gives an insight into the state of health of the infant and ...

  19. Multiple nuclear dots and rim-like/membranous IgG isotypes in primary biliary cirrhosis.

    Science.gov (United States)

    Muratori, Paolo; Granito, Alessandro; Ferri, Silvia; Pappas, Georgios; Volta, Umberto; Menichella, Rita; Bianchi, Francesco B; Lenzi, Marco; Muratori, Luigi

    2009-03-01

    Anti nuclear (ANA) immunomorphological patterns such as multiple nuclear dots (MND) and rim-like/membranous (RL/M) are considered highly specific but little sensitive for primary biliary cirrhosis (PBC) diagnosis. To evaluate frequency and clinical significance of MND and RL/M in PBC patients when investigated at the level of immunoglobulin G isotypes. MND and RL/M pattern have been tested in 141 PBC sera and 230 pathological controls using HEp-2 cells as substrate and anti- total IgG and individual IgG subclasses (IgG1, IgG2, IgG3, IgG4) as specific antisera. One hundred and fourteen of 141 (80%) PBC patients had RL/M or MND pattern when IgG subclasses were used as revealing reagents (vs. 34% when anti total IgG were used, p < 0.0001). The prevalent isotype was IgG1 for RL/M, and IgG2 for MND pattern. None of controls was positive. No clinical differences in terms of severity and outcome of disease have been observed in PBC patients positive for MND and RL/M when investigated with IgG isotypes. The research for RL/M and MND pattern at level of IgG isotype determines a wide gain in terms of sensitivity without a loss of specificity. In Italian PBC patients MND and RL/M pattern did not seem to characterize any subgroup of patients with a poorer prognosis.

  20. Mitochondrial-dependent Autoimmunity in Membranous Nephropathy of IgG4-related Disease

    Directory of Open Access Journals (Sweden)

    Simona Buelli

    2015-05-01

    Full Text Available The pathophysiology of glomerular lesions of membranous nephropathy (MN, including seldom-reported IgG4-related disease, is still elusive. Unlike in idiopathic MN where IgG4 prevails, in this patient IgG3 was predominant in glomerular deposits in the absence of circulating anti-phospholipase A2 receptor antibodies, suggesting a distinct pathologic process. Here we documented that IgG4 retrieved from the serum of our propositus reacted against carbonic anhydrase II (CAII at the podocyte surface. In patient's biopsy, glomerular CAII staining increased and co-localized with subepithelial IgG4 deposits along the capillary walls. Patient's IgG4 caused a drop in cell pH followed by mitochondrial dysfunction, excessive ROS production and cytoskeletal reorganization in cultured podocytes. These events promoted mitochondrial superoxide-dismutase-2 (SOD2 externalization on the plasma membrane, becoming recognizable by complement-binding IgG3 anti-SOD2. Among patients with IgG4-related disease only sera of those with IgG4 anti-CAII antibodies caused low intracellular pH and mitochondrial alterations underlying SOD2 externalization. Circulating IgG4 anti-CAII can cause podocyte injury through processes of intracellular acidification, mitochondrial oxidative stress and neoantigen induction in patients with IgG4 related disease. The onset of MN in a subset of patients could be due to IgG4 antibodies recognizing CAII with consequent exposure of mitochondrial neoantigen in the context of multifactorial pathogenesis of disease.

  1. The ue of polysiloxane/polyvinyl alcohol beads as solid phase in IgG anti-Toxocara canis detection using a recombinant antigen

    Directory of Open Access Journals (Sweden)

    Raquel de Andrade Lima Coêlho

    2003-04-01

    Full Text Available Immunodetection of human IgG anti-Toxocara canis was developed based on ELISA and on the use of polysiloxane/polyvinyl alcohol (POS/PVA beads. A recombinant antigen was covalently immobilized, via glutaraldehyde, onto this hybrid inorganic-organic composite, which was prepared by the sol-gel technique. Using only 31.2 ng antigen per bead, a peroxidase conjugate dilution of 1:10,000 and a serum dilution of 1:200 were adequate for the establishment of the procedure. This procedure is comparable to that which utilizes the adsorption of the antigen to conventional PVC plates. However, the difference between positive and negative sera mean absorbances was larger for this new glass based assay. In addition to the performance of the POS/PVA bead as a matrix for immunodetection, its easy synthesis and low cost are additional advantages for commercial application.

  2. Kinetics of IgG antibody to cytomegalovirus (CMV) after birth and seroprevalence of anti-CMV IgG in Chinese children

    OpenAIRE

    Chen, Jie; Hu, Lingqing; Wu, Meiling; Zhong, Tianying; Zhou, Yi-Hua; Hu, Yali

    2012-01-01

    Abstract Background Prevalence of cytomegalovirus (CMV) infection is 90–100% in developing countries; however, the kinetics of anti-CMV IgG in infants remains elusive. Methods Sera from 112 mother-newborn pairs and longitudinal samples from 41 infants up to 2-year old were tested for anti-CMV IgG and IgM. Additionally, samples from 837 healthy children were included. Results Of 112 mothers, 108 (96.4%) were anti-CMV IgG positive; their 108 newborns were also seropositive. In a 2-year follow-u...

  3. ELISA for Detection of Soya Proteins in Meat Products

    Directory of Open Access Journals (Sweden)

    Eva Renčová

    2009-01-01

    Full Text Available Indirect competitive ELISA method for the detection of soya proteins in meat products was developed. The detection limit of the method is 0.5% of the weight of added soya protein. A total of 131 meat product samples such as salamis or sausages from the Czech Republic market were investigated for the presence of soya proteins. Soya proteins were detected in 84% of the investigated samples without any declaration on the package of the product. The use of vegetable additives, namely soya in meat products in the market of the Czech Republic is very frequent and the restriction of its usage by legislation relates only to some kinds of durable products and ham (Act 264/2003 Coll.. The need for sensitive inspecting methods for soya protein detection is not only associated with the economic aspect (adulteration, but mainly with consumer health protection in case of allergy to soya proteins.

  4. IgG4-related Disease - A Systemic Disease that Deserves Attention Regardless of One's Subspecialty.

    Science.gov (United States)

    Hamano, Hideaki; Tanaka, Eiji; Ishizaka, Nobukazu; Kawa, Shigeyuki

    2017-12-27

    IgG4-related disease (IgG4-RD) is an inflammatory condition characterized by a high serum IgG4 concentration and the abundant infiltration of lymphocytes and IgG4-positive plasma cells in the tissue, as well as spatial (diverse clinical manifestations) and temporal (the possibility of recurrence) multiplicities. Since the initial documentation of IgG4-related disease in patients with autoimmune pancreatitis in 2001, a growing body of evidence has been accumulating to suggest that various-virtually all-organs can be affected by IgG4-RD. In general, steroid therapy is effective and is considered to be the first-line treatment for IgG4-RD. The precise mechanism underlying this systemic disorder has remained unknown. Considering that IgG4-RD was specified as being an intractable disease in 2015, further studies are needed to clarify whether IgG4-RD is indeed a distinct disease entity or a complex of disorders of different etiologies and clinical conditions.

  5. Clinical and pathological characteristics of IgG4-related interstitial lung disease

    Science.gov (United States)

    Lv, Xiaoting; Gao, Feng; Liu, Qicai; Zhang, Sheng; Huang, Zhihua; Zhu, Yongping; Zong, Haiyang; Li, Quwen; Li, Sanyan

    2018-01-01

    IgG4-related interstitial lung disease (IgG4-RILD), which is characterized by increased IgG4 levels, IgG4+ plasma cell infiltration and irregular whorled fibrosis, is a recently described lung disorder that belongs to the group of systemic fibroinflammatory IgG4-related diseases (IgG4-RD). The aim of the present study was to improve the current knowledge regarding the specific clinical and histopathological characteristics of IgG4-RILD and to investigate its underlying immune mechanism in vivo. A total of 7 patients newly diagnosed with IgG4-RILD were enrolled in the present study (4 men and 3 women; mean age, 57 years; range, 29–71 years). Patients' clinical history was collected and serological indicators, including C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), antinuclear antibodies (ANAs) and tumor markers were measured. Serum immunoglobulin G (IgG), IgE and IgG4 levels were also evaluated. In addition, computed tomographic (CT) images and pathological examinations were used to determine the characteristics of lung lesions in all patients. The majority of patients presented with symptoms of fever, cough and dyspnea, while allergic symptoms were also encountered. The laboratory examination results revealed different degrees of increased CRP, ESR, tumor markers, ANA, serum IgE and IgG4. The CT images revealed diffuse ground glass opacities, bronchiectasis and thickened bronchovascular bundles. Histologically, the lung lesions were characterized by dense IgG4+ lymphoplasmacytic infiltrates intermixed with extensive fibrous tissue hyperplasia and an irregularly storiform pattern of fibrosis. The mean number of IgG4+ plasma cells was >10 cells/high power field. The ratio of IgG/IgG4+ plasma cells was >50% in inflamed lesions and the number of parenchymal cells was markedly reduced. Obliterative phlebitis or obliterative arteritis was observed in all patients. In conclusion, the clinicopathological similarities between IgG4-RILD and other IgG4-RD

  6. IgG subclass and vaccination stimulus determine changes in antigen specific antibody glycosylation in mice.

    Science.gov (United States)

    Kao, Daniela; Lux, Anja; Schaffert, Anja; Lang, Roland; Altmann, Friedrich; Nimmerjahn, Falk

    2017-12-01

    Immunoglobulin G (IgG) glycosylation can modulate antibody effector functions. Depending on the precise composition of the sugar moiety attached to individual IgG glycovariants either pro- or anti-inflammatory effector pathways can be initiated via differential binding to type I or type II Fc-receptors. However, an in depth understanding of how individual IgG subclasses are glycosylated during the steady state and how their glycosylation pattern changes during vaccination is missing. To monitor IgG subclass glycosylation during the steady state and upon vaccination of mice with different T-cell dependent and independent antigens, tryptic digests of serum, and antigen-specific IgG preparations were analyzed by reversed phase-liquid chromatography-mass spectrometry. We show that there is a remarkable difference with respect to how individual IgG subclasses are glycosylated during the steady state. More importantly, upon T-cell dependent and independent vaccinations, individual antigen-specific IgG subclasses reacted differently with respect to changes in individual glycoforms, suggesting that the IgG subclass itself is a major determinant of restricting or allowing alterations in specific IgG glycovariants. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Community Rates of IgG4 Antibodies to Ascaris Haemoglobin Reflect Changes in Community Egg Loads Following Mass Drug Administration

    Science.gov (United States)

    Vlaminck, Johnny; Supali, Taniawati; Geldhof, Peter; Hokke, Cornelis H.; Fischer, Peter U.; Weil, Gary J.

    2016-01-01

    Background Conventional diagnostic methods for human ascariasis are based on the detection of Ascaris lumbricoides eggs in stool samples. However, studies of ascariasis in pigs have shown that the prevalence and the number of eggs detected in the stool do not correlate well with exposure of the herd to the parasite. On the other hand, an ELISA test measuring antibodies to Ascaris suum haemoglobin (AsHb) has been shown to be useful for estimating transmission intensity on pig farms. In this study, we further characterized the AsHb antigen and screened samples from a population-based study conducted in an area that is endemic for Ascaris lumbricoides in Indonesia to assess changes in AsHb antibody rates and levels in humans following mass drug administration (MDA). Methodology/Principal findings We developed and evaluated an ELISA to detect human IgG4 antibodies to AsHb. We tested 1066 plasma samples collected at different times from 599 subjects who lived in a village in rural Indonesia that was highly endemic for ascariasis. The community received 6 rounds of MDA for lymphatic filariasis with albendazole plus diethylcarbamazine between 2002 and 2007. While the AsHb antibody assay was not sensitive for detecting all individuals with Ascaris eggs in their stools, the percentage of seropositive individuals decreased rapidly following MDA. Reductions in antibody rates reflected decreased mean egg output per person both at the community level and in different age groups. Two years after the last round of MDA the community egg output and antibody prevalence rate were reduced by 81.6% and 78.9% respectively compared to baseline levels. Conclusion/Significance IgG4 antibody levels to AsHb appear to reflect recent exposure to Ascaris. The antibody prevalence rate may be a useful indicator for Ascaris transmission intensity in communities that can be used to assess the impact of control measures on the force of transmission. PMID:26991326

  8. Usefulness of FC-TRIPLEX Chagas/Leish IgG1 as confirmatory assay for non-negative results in blood bank screening of Chagas disease.

    Science.gov (United States)

    Campos, Fernanda Magalhães Freire; Repoles, Laura Cotta; de Araújo, Fernanda Fortes; Peruhype-Magalhães, Vanessa; Xavier, Marcelo Antônio Pascoal; Sabino, Ester Cerdeira; de Freitas Carneiro Proietti, Anna Bárbara; Andrade, Mariléia Chaves; Teixeira-Carvalho, Andréa; Martins-Filho, Olindo Assis; Gontijo, Célia Maria Ferreira

    2018-04-01

    A relevant issue in Chagas disease serological diagnosis regards the requirement of using several confirmatory methods to elucidate the status of non-negative results from blood bank screening. The development of a single reliable method may potentially contribute to distinguish true and false positive results. Our aim was to evaluate the performance of the multiplexed flow-cytometry anti-T. cruzi/Leishmania IgG1 serology/(FC-TRIPLEX Chagas/Leish IgG1) with three conventional confirmatory criteria (ELISA-EIA, Immunofluorescence assay-IIF and EIA/IIF consensus criterion) to define the final status of samples with actual/previous non-negative results during anti-T. cruzi ELISA-screening in blood banks. Apart from inconclusive results, the FC-TRIPLEX presented a weak agreement index with EIA, while a strong agreement was observed when either IIF or EIA/IIF consensus criteria were applied. Discriminant analysis and Spearman's correlation further corroborates the agreement scores. ROC curve analysis showed that FC-TRIPLEX performance indexes were higher when IIF and EIA/IIF consensus were used as a confirmatory criterion. Logistic regression analysis further demonstrated that the probability of FC-TRIPLEX to yield positive results was higher for inconclusive results from IIF and EIA/IIF consensus. Machine learning tools illustrated the high level of categorical agreement between FC-TRIPLEX versus IIF or EIA/IIF consensus. Together, these findings demonstrated the usefulness of FC-TRIPLEX as a tool to elucidate the status of non-negative results in blood bank screening of Chagas disease. Copyright © 2018. Published by Elsevier B.V.

  9. Community Rates of IgG4 Antibodies to Ascaris Haemoglobin Reflect Changes in Community Egg Loads Following Mass Drug Administration.

    Science.gov (United States)

    Vlaminck, Johnny; Supali, Taniawati; Geldhof, Peter; Hokke, Cornelis H; Fischer, Peter U; Weil, Gary J

    2016-03-01

    Conventional diagnostic methods for human ascariasis are based on the detection of Ascaris lumbricoides eggs in stool samples. However, studies of ascariasis in pigs have shown that the prevalence and the number of eggs detected in the stool do not correlate well with exposure of the herd to the parasite. On the other hand, an ELISA test measuring antibodies to Ascaris suum haemoglobin (AsHb) has been shown to be useful for estimating transmission intensity on pig farms. In this study, we further characterized the AsHb antigen and screened samples from a population-based study conducted in an area that is endemic for Ascaris lumbricoides in Indonesia to assess changes in AsHb antibody rates and levels in humans following mass drug administration (MDA). We developed and evaluated an ELISA to detect human IgG4 antibodies to AsHb. We tested 1066 plasma samples collected at different times from 599 subjects who lived in a village in rural Indonesia that was highly endemic for ascariasis. The community received 6 rounds of MDA for lymphatic filariasis with albendazole plus diethylcarbamazine between 2002 and 2007. While the AsHb antibody assay was not sensitive for detecting all individuals with Ascaris eggs in their stools, the percentage of seropositive individuals decreased rapidly following MDA. Reductions in antibody rates reflected decreased mean egg output per person both at the community level and in different age groups. Two years after the last round of MDA the community egg output and antibody prevalence rate were reduced by 81.6% and 78.9% respectively compared to baseline levels. IgG4 antibody levels to AsHb appear to reflect recent exposure to Ascaris. The antibody prevalence rate may be a useful indicator for Ascaris transmission intensity in communities that can be used to assess the impact of control measures on the force of transmission.

  10. Hybrid radioguided occult lesion localization (hybrid ROLL) of (18)F-FDG-avid lesions using the hybrid tracer indocyanine green-(99m)Tc-nanocolloid.

    Science.gov (United States)

    KleinJan, G H; Brouwer, O R; Mathéron, H M; Rietbergen, D D D; Valdés Olmos, R A; Wouters, M W; van den Berg, N S; van Leeuwen, F W B

    2016-01-01

    To assess if combined fluorescence- and radio-guided occult lesion localization (hybrid ROLL) is feasible in patients scheduled for surgical resection of non-palpable (18)F-FDG-avid lesions on PET/CT. Four patients with (18)F-FDG-avid lesions on follow-up PET/CT that were not palpable during physical examination but were suspected to harbor metastasis were enrolled. Guided by ultrasound, the hybrid tracer indocyanine green (ICG)-(99m)Tc-nanocolloid was injected centrally in the target lesion. SPECT/CT imaging was used to confirm tracer deposition. Intraoperatively, lesions were localized using a hand-held gamma ray detection probe, a portable gamma camera, and a fluorescence camera. After excision, the gamma camera was used to check the wound bed for residual activity. A total of six (18)F-FDG-avid lymph nodes were identified and scheduled for hybrid ROLL. Comparison of the PET/CT images with the acquired SPECT/CT after hybrid tracer injection confirmed accurate tracer deposition. No side effects were observed. Combined radio- and fluorescence-guidance enabled localization and excision of the target lesion in all patients. Five of the six excised lesions proved tumor-positive at histopathology. The hybrid ROLL approach appears to be feasible and can facilitate the intraoperative localization and excision of non-palpable lesions suspected to harbor tumor metastases. In addition to the initial radioguided detection, the fluorescence component of the hybrid tracer enables high-resolution intraoperative visualization of the target lesion. The procedure needs further evaluation in a larger cohort and wider range of malignancies to substantiate these preliminary findings. Copyright © 2016 Elsevier España, S.L.U. y SEMNIM. All rights reserved.

  11. Vesicular monoamine transporter protein expression correlates with clinical features, tumor biology, and MIBG avidity in neuroblastoma: a report from the Children's Oncology Group

    Energy Technology Data Exchange (ETDEWEB)

    Temple, William; Mendelsohn, Lori; Nekritz, Erin; Gustafson, W.C.; Matthay, Katherine K. [UCSF School of Medicine, Department of Pediatrics, San Francisco, CA (United States); UCSF Benioff Children' s Hospital, San Francisco, CA (United States); Kim, Grace E. [UCSF School of Medicine, Department of Pathology, San Francisco, CA (United States); Lin, Lawrence; Giacomini, Kathy [UCSF School of Pharmacy, Department of Bioengineering and Therapeutic Sciences, San Francisco, CA (United States); Naranjo, Arlene; Van Ryn, Collin [University of Florida, Children' s Oncology Group Statistics and Data Center, Gainesville, FL (United States); Yanik, Gregory A. [University of Michigan, CS Mott Children' s Hospital, Ann Arbor, MI (United States); Kreissman, Susan G. [Duke University Medical Center, Durham, NC (United States); Hogarty, Michael [University of Pennsylvania, Children' s Hospital of Philadelphia and Perelman School of Medicine, Philadelphia, PA (United States); DuBois, Steven G. [UCSF School of Medicine, Department of Pediatrics, San Francisco, CA (United States); UCSF Benioff Children' s Hospital, San Francisco, CA (United States); UCSF School of Medicine, San Francisco, CA (United States)

    2016-03-15

    Vesicular monoamine transporters 1 and 2 (VMAT1 and VMAT2) are thought to mediate MIBG uptake in adult neuroendocrine tumors. In neuroblastoma, the norepinephrine transporter (NET) has been investigated as the principal MIBG uptake protein, though some tumors without NET expression concentrate MIBG. We investigated VMAT expression in neuroblastoma and correlated expression with MIBG uptake and clinical features. We evaluated VMAT1 and VMAT2 expression by immunohistochemistry (IHC) in neuroblastoma tumors from 76 patients with high-risk metastatic disease treated in a uniform cooperative group trial (COG A3973). All patients had baseline MIBG diagnostic scans centrally reviewed. IHC results were scored as the product of intensity grading (0 - 3+) and percent of tumor cells expressing the protein of interest. The association between VMAT1 and VMAT2 scores and clinical and biological features was tested using Wilcoxon rank-sum tests. Patient characteristics were typical of high-risk neuroblastoma, though the cohort was intentionally enriched in patients with MIBG-nonavid tumors (n = 20). VMAT1 and VMAT2 were expressed in 62 % and 75 % of neuroblastoma tumors, respectively. VMAT1 and VMAT2 scores were both significantly lower in MYCN amplified tumors and in tumors with high mitotic karyorrhectic index. MIBG-avid tumors had significantly higher VMAT2 scores than MIBG-nonavid tumors (median 216 vs. 45; p = 0.04). VMAT1 expression did not correlate with MIBG avidity. VMAT1 and VMAT2 are expressed in the majority of neuroblastomas. Expression correlates with other biological features. The expression level of VMAT2 but not that of VMAT1 correlates with avidity for MIBG. (orig.)

  12. Immunogenetics of IgG4-Related AIP.

    Science.gov (United States)

    Ota, Masao; Umemura, Takeji; Kawa, Shigeyuki

    2017-01-01

    Autoimmune pancreatitis (AIP) is a unique form of chronic pancreatitis characterized by high serum IgG4 concentration and a variety of complicating extra-pancreatic lesions. AIP has the features of a complex disease that is caused by multifactorial genes. However, the genetic factors underlying AIP have not been elucidated conclusively. Association studies by the candidate-gene approach and genome-wide association studies (GWAS) have revealed several susceptibility genes for AIP, including HLA DRB1*04:05-DQB1*04:01, FCRL3, CTLA4, and KCNA3, albeit in small-scale analyses. Thus, GWAS of large sample sizes and multinational collaborative meta-analyses are needed to identify the precise genetic variants that are associated with AIP onset. Systems genetics approaches that integrate DNA sequencing, expression quantitative trait locus (eQTL) mapping, proteomics, and metabolomics will also be useful in clarifying the pathogenesis of AIP.

  13. Fc-Glycosylation in Human IgG1 and IgG3 Is Similar for Both Total and Anti-Red-Blood Cell Anti-K Antibodies

    Directory of Open Access Journals (Sweden)

    Myrthe E. Sonneveld

    2018-01-01

    Full Text Available After albumin, immunoglobulin G (IgG are the most abundant proteins in human serum, with IgG1 and IgG3 being the most abundant subclasses directed against protein antigens. The quality of the IgG-Fc-glycosylation has important functional consequences, which have been found to be skewed toward low fucosylation in some antigen-specific immune responses. This increases the affinity to IgG1-Fc-receptor (FcγRIIIa/b and thereby directly affects downstream effector functions and disease severity. To date, antigen-specific IgG-glycosylation have not been analyzed for IgG3. Here, we analyzed 30 pregnant women with anti-K alloantibodies from a prospective screening cohort and compared the type of Fc-tail glycosylation of total serum- and antigen-specific IgG1 and IgG3 using mass spectrometry. Total serum IgG1 and IgG3 Fc-glycoprofiles were highly similar. Fc glycosylation of antigen-specific IgG varied greatly between individuals, but correlated significantly with each other for IgG1 and IgG3, except for bisection. However, although the magnitude of changes in fucosylation and galactosylation were similar for both subclasses, this was not the case for sialylation levels, which were significantly higher for both total and anti-K IgG3. We found that the combination of relative IgG1 and IgG3 Fc-glycosylation levels did not improve the prediction of anti-K mediated disease over IgG1 alone. In conclusion, Fc-glycosylation profiles of serum- and antigen-specific IgG1 and IgG3 are highly similar.

  14. Monitoring the systemic human memory B cell compartment of melanoma patients for anti-tumor IgG antibodies.

    Directory of Open Access Journals (Sweden)

    Amy E Gilbert

    Full Text Available Melanoma, a potentially lethal skin cancer, is widely thought to be immunogenic in nature. While there has been much focus on T cell-mediated immune responses, limited knowledge exists on the role of mature B cells. We describe an approach, including a cell-based ELISA, to evaluate mature IgG antibody responses to melanoma from human peripheral blood B cells. We observed a significant increase in antibody responses from melanoma patients (n = 10 to primary and metastatic melanoma cells compared to healthy volunteers (n = 10 (P<0.0001. Interestingly, we detected a significant reduction in antibody responses to melanoma with advancing disease stage in our patient cohort (n = 21 (P<0.0001. Overall, 28% of melanoma patient-derived B cell cultures (n = 1,800 compared to 2% of cultures from healthy controls (n = 600 produced antibodies that recognized melanoma cells. Lastly, a patient-derived melanoma-specific monoclonal antibody was selected for further study. This antibody effectively killed melanoma cells in vitro via antibody-mediated cellular cytotoxicity. These data demonstrate the presence of a mature systemic B cell response in melanoma patients, which is reduced with disease progression, adding to previous reports of tumor-reactive antibodies in patient sera, and suggesting the merit of future work to elucidate the clinical relevance of activating humoral immune responses to cancer.

  15. Monitoring the systemic human memory B cell compartment of melanoma patients for anti-tumor IgG antibodies.

    Science.gov (United States)

    Gilbert, Amy E; Karagiannis, Panagiotis; Dodev, Tihomir; Koers, Alexander; Lacy, Katie; Josephs, Debra H; Takhar, Pooja; Geh, Jenny L C; Healy, Ciaran; Harries, Mark; Acland, Katharine M; Rudman, Sarah M; Beavil, Rebecca L; Blower, Philip J; Beavil, Andrew J; Gould, Hannah J; Spicer, James; Nestle, Frank O; Karagiannis, Sophia N

    2011-04-29

    Melanoma, a potentially lethal skin cancer, is widely thought to be immunogenic in nature. While there has been much focus on T cell-mediated immune responses, limited knowledge exists on the role of mature B cells. We describe an approach, including a cell-based ELISA, to evaluate mature IgG antibody responses to melanoma from human peripheral blood B cells. We observed a significant increase in antibody responses from melanoma patients (n = 10) to primary and metastatic melanoma cells compared to healthy volunteers (n = 10) (P<0.0001). Interestingly, we detected a significant reduction in antibody responses to melanoma with advancing disease stage in our patient cohort (n = 21) (P<0.0001). Overall, 28% of melanoma patient-derived B cell cultures (n = 1,800) compared to 2% of cultures from healthy controls (n = 600) produced antibodies that recognized melanoma cells. Lastly, a patient-derived melanoma-specific monoclonal antibody was selected for further study. This antibody effectively killed melanoma cells in vitro via antibody-mediated cellular cytotoxicity. These data demonstrate the presence of a mature systemic B cell response in melanoma patients, which is reduced with disease progression, adding to previous reports of tumor-reactive antibodies in patient sera, and suggesting the merit of future work to elucidate the clinical relevance of activating humoral immune responses to cancer.

  16. Influenza A haemagglutinin specific IgG responses in children and adults after seasonal trivalent live attenuated influenza vaccination.

    Science.gov (United States)

    Islam, Shahinul; Mohn, Kristin Greve-Isdahl; Krammer, Florian; Sanne, Mari; Bredholt, Geir; Jul-Larsen, Åsne; Tete, Sarah M; Zhou, Fan; Brokstad, Karl Albert; Cox, Rebecca Jane

    2017-10-09

    Influenza is a major respiratory pathogen and vaccination is the main method of prophylaxis. In 2012, the trivalent live attenuated influenza vaccine (LAIV3) was licensed in Europe for use in children. Vaccine-induced antibodies directed against the main viral surface glycoprotein, haemagglutinin (HA), play an important role in virus neutralization through different mechanism. The objective of this study was to dissect the HA specific antibody responses induced after LAIV3 immunization to the influenza A viruses in children and adults. Plasma was collected from 20 children and 20 adults pre- and post-LAIV3 vaccination (up to ayear) and analysed by the haemagglutination inhibition (HI) and ELISA assays. We found that LAIV3 boosted the HA specific IgG response against the head and the full-length of H3N2 in children, but not adults. Adults had higher levels of pre-existing stalk antibodies (towards H3N2 and H1N1), but these were not boosted by LAIV3. Importantly, we observed a trend in boosting of H1N1 HA stalk specific antibodies in children after LAIV3. Whereas, heterosubtypic H5 and H7 full-length HA specific antibodies were not boosted in either children or adults. In conclusion, LAIV3 elicited H3-head and low levels of H1 stalk specific antibody responses in children, supporting the prophylactic use of LAIV in children. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Prevalence of IgG Antibodies to Toxoplasma gondii in Veterinary and Undergraduate Students at Virginia Tech, Blacksburg, Virginia.

    Science.gov (United States)

    Rosypal, A C; Houk, A E; Zajac, A M; Lindsay, D S

    2015-11-01

    Toxoplasma gondii is a globally distributed parasitic protozoan that infects humans and other warm-blooded vertebrates. Felids are the only definitive host for T. gondii, and they excrete oocysts in their faeces. The national prevalence in humans is declining in the United States. This zoonotic organism is of particular interest due to its importance in pregnant women, in individuals with altered immune systems, and in reactivated ocular infections. Exposure to the parasite in humans is usually associated with consumption of raw or undercooked meat or by accidental ingestion of oocysts. It was hypothesized that veterinary students would have a greater chance at exposure to the parasite than an average population of undergraduate students due to increased contact with cats who are infected. A commercially available ELISA was used to examine serum samples from 336 students (252 veterinary students and 84 undergraduate students) at Virginia Polytechnic Institute and State University and the Virginia-Maryland Regional College of Veterinary Medicine for serum IgG antibodies to T. gondii antigen. The prevalence of T. gondii in these subjects was 5.6% in veterinary school students (n = 252) and 2.4% in undergraduates (n = 84). There was no significant difference (P > 0.05) in the prevalence of T. gondii antibodies in veterinary versus undergraduate students. The overall prevalence of 4.8% in all students in this study reflects the continuing decline of antibodies to T. gondii in humans in the United States. © 2015 Blackwell Verlag GmbH.

  18. A myoglobin mutant designed to mimic the oxygen-avid Ascaris suum hemoglobin: elucidation of the distal hydrogen bonding network by solution NMR.

    OpenAIRE

    Zhang, W; Cutruzzolá, F; Allocatelli, C T; Brunori, M; La Mar, G N

    1997-01-01

    The solution 1H NMR structure of the active site and ligand dissociation rate for the cyanomet complex have been determined for a sperm whale myoglobin triple mutant Leu29(B10)-->Tyr, His64(E7)-->Gln, Thr67(E10)-->Arg that mimics the distal residue configuration of the oxygen-avid hemoglobin from Ascaris suum. A double mutant that retains Leu29(B10) was similarly investigated. Two-dimensional NMR analysis of the iron-induced dipolar shifts, together with the conserved proximal side structure ...

  19. Comparison among the BED capture enzyme immunoassay test and AxSYM avidity index assay for determining recent HIV infection and incidence in two Voluntary Counselling and Testing Centres in Northeast Brazil

    Directory of Open Access Journals (Sweden)

    Daniela Medeiros Salustiano

    2014-07-01

    Full Text Available The aims of this study were to compare the automated AxSYM avidity assay index with the BED capture enzyme immunoassay test and to calculate the HIV-1 incidence using the BED capture enzyme immunoassay and AxSYM avidity assay index algorithms within a population seeking the Voluntary Counselling and Testing Centres in two municipalities in the Metropolitan Region of Recife, Northeast of Brazil. An analysis was conducted in 365 samples that tested positive for HIV infection from frozen serum collected during the period 2006–2009. There was a similar proportion of males and females; most patients were heterosexual (86% with a median age of 29 years. Of the 365 samples, 102 (28% and 66 (18.1% were identified as recent infections by BED capture enzyme immunoassay and AxSYM avidity assay index, respectively. The HIV-1 total incidence in the BED capture enzyme immunoassay and AxSYM avidity assay index algorithms were: 0.79 (95% CI: 0.60–0.98 and 0.34 (95% CI: −0.04 to 0.72, respectively. Incidence was higher among men. There was good agreement between the tests, with a kappa of 0.654 and a specificity of 95.8%. AxSYM avidity assay index may be helpful in improving the quality of the estimates of recent HIV infection and incidence, particularly when used in a combined algorithm with BED capture enzyme immunoassay.

  20. Pemphigus vulgaris is characterized by low IgG reactivities to specific self-antigens along with high IgG reactivity to desmoglein 3

    Science.gov (United States)

    Fattal, Ittai; Rimer, Jacob; Shental, Noam; Molad, Yair; Gabrielli, Armando; Livneh, Avi; Sarig, Ofer; Goldberg, Ilan; Gafter, Uzi; Domany, Eytan; Cohen, Irun R

    2014-01-01

    Pemphigus vulgaris (PV) is an autoimmune skin disease, which has been characterized by IgG autoantibodies to desmoglein 3. Here we studied the antibody signatures of PV patients compared with healthy subjects and with patients with two other autoimmune diseases with skin manifestations (systemic lupus erythematosus and scleroderma), using an antigen microarray and informatics analysis. We now report a previously unobserved phenomenon – patients with PV, compared with the healthy subjects and the two other diseases, show a significant decrease in IgG autoantibodies to a specific set of self-antigens. This novel finding demonstrates that an autoimmune disease may be associated with a loss of specific, healthy IgG autoantibodies and not only with a gain of specific, pathogenic IgG autoantibodies. PMID:24820664

  1. IgG4-related skin disease may have distinct systemic manifestations: a systematic review.

    Science.gov (United States)

    Bennett, Adam E; Fenske, Neil A; Rodriguez-Waitkus, Paul; Messina, Jane L

    2016-11-01

    IgG4-related disease (IgG4-RD) is an increasingly prevalent protean multisystem disorder characterized by single or multi-organ infiltration of IgG4-bearing plasma cells. Skin involvement has been recognized and is relevant to proper diagnosis. A systematic literature review of 50 cases involving the skin reveals that patients with IgG4-related skin disease show predominant involvement of the head and neck and have a distinct pattern of systemic involvement, also favoring the head and neck - lymphatics, orbit, salivary, and lacrimal glands - but generally lacking pancreaticobiliary involvement (16% of cases), which by contrast is a predominant manifestation in systemic IgG4-RD (60% with pancreaticobiliary involvement). We summarize clinical and pathologic descriptive data from this systematic review. We review differential diagnosis and propose a diagnostic scheme for stratifying probability of disease based upon comprehensive integration of clinical, histopathologic, and laboratory data. Plasmacyte infiltration and storiform fibrosis are prominent in IgG4-related skin disease, but obliterative venulitis is less common than in the prototypical IgG4-related disease manifestation of autoimmune pancreatitis. IgG4 tissue and serum values, with a mean (±95% CI) in the reviewed cases of 132.8 ± 32.6 IgG4-positive plasma cells per high-power field and 580 ± 183.8 mg/dl, respectively, are incorporated into the suggested criteria. The distinct set of manifestations identified by this systematic review and the proposed diagnostic considerations, while requiring further validation in prospective studies, highlight the need to consider that IgG4-related skin disease defines a unique systemic disease complex along the spectrum of IgG4-RD. © 2016 The International Society of Dermatology.

  2. Cloning of pCDNA3-IgG4 and pQE-2-IgG4 human hinge region ...

    African Journals Online (AJOL)

    GREGORY

    2011-12-16

    Dec 16, 2011 ... peptide should comprise of 68 residues of the expected amino acid sequence. Construction of pQE-2-IgG4 expression plasmid DNA. Figure 2 shows the agarose gel electrophoresis result of restriction digestion of pQE-2-IgG4 with Not1 and Nde1. Two bands representing the vector pQE-2 (4.8 kb) and.

  3. Cloning of pCDNA3-IgG4 and pQE-2-IgG4 human hinge region ...

    African Journals Online (AJOL)

    The present study was conducted to prepare the plasmid construct of pQE-2-IgG4 for peptide expression and pCDNA3-IgG4 for use in intrasplenic immunization in view of monoclonal antibody production. pQE-2 is a prokaryotic expression vector whereas pCDNA3 is a mammalian expression vector. Some methods were ...

  4. Deficiency of IgG4 in children: association of isolated IgG4 deficiency with recurrent respiratory tract infection.

    Science.gov (United States)

    Moss, R B; Carmack, M A; Esrig, S

    1992-01-01

    To study the relationship between serum IgG subclass deficiency and clinical host defense impairment, we reviewed the clinical and immunologic features of 123 patients with a history of recurrent infection who had been examined for immunodeficiency in our laboratory (group 1). We then compared immunoglobulin isotype levels with those in sera from 127 age-matched control subjects without recurrent infection from whom blood had been drawn for evaluation of atopy (group 2). There was a significantly higher prevalence of IgG4 deficiencies among patients with recurrent infections (17% vs 7%; p less than 0.02), solely because of a higher prevalence of isolated IgG4 deficiency (n = 9; 7.3%) than in atopic control subjects (n = 1; 0.8%; p less than 0.05); there was a comparable prevalence of multiple isotype deficiencies that included low levels of IgG4 (9.8% and 6.3%, respectively). All nine group 1 patients with isolated IgG4 deficiency had severe recurrent respiratory tract infections requiring multiple hospitalizations; in addition, five were atopic, five had asthma, and one had chronic diahrrea. Antibody responses to bacterial polysaccharide antigens were normal for age in all patients with isolated IgG4 deficiency; two had defective antibody responses to protein antigens. Isolated IgG4 deficiency appears to be associated with impaired respiratory tract defenses and may occur in the absence of an easily definable antibody deficiency state. This association suggests a physiologic defense role for mucosal IgG4.

  5. Evaluation of an indirect ELISA for detection and typing of foot-and-mouth disease virus

    International Nuclear Information System (INIS)

    Prado, J.A.

    1998-01-01

    An indirect enzyme linked immunosorbent assay (ELISA) kit was used for diagnosis of foot-and-mouth disease virus (FMDV) types O1, A23, C3 which occurred in Rio Grande do Sul State, Southern Brazil during 1984-1994. The samples were randomly selected and tested by ELISA, Complement Fixation Test (CFT) and in tissue culture. Out of 106 samples 78 (73,5%) were positive by ELISA and 39 (36,8%) were found positive in CFT, when original suspensions were used. Once these samples were inoculated onto tissue culture both tests gave similar results, although ELISA picked up more positive samples during the 1st passage in tissue culture. The negative samples (16) included in this study were negative in all tests. The ELISA was more sensitive than and as specific as CFT. ELISA and tissue culture together were shown to be a better system for detection of foot-and-mouth disease virus antigen than CFT. (author)

  6. [A double antibody sandwich ELISA based assay for titration of severe fever with thrombocytopenia syndrome virus].

    Science.gov (United States)

    Liu, Lin; Zhang, Quan-Fu; Li, Chuan; Li, Jian-Dong; Jiang, Xiao-Lin; Zhang, Fu-Shun; Wu, Wei; Liang, Mi-Fang; Li, De-Xin

    2013-06-01

    To develop an assay for titration of severe fever with thrombocytopenia syndrome virus (SFTSV) based on double antibody sandwich ELISA. A double antibody sandwich ELISA was developed for detection of SFTSV based on SFTSV nucleocapsid (N) protein specific poly- and monoclonal antibodies, procedures were optimized and evaluated. This ELISA based titration assay was compared with fluorescence assasy and plaque assay based titration method. The results suggested that the titers obtained by ELISA based method are consistent with those obtained by IFA based method (R = 0.999) and the plaque assay titration method (R = 0.949). The novel ELISA based titration method with high sensitivity and specificity is easy to manage and perform, and can overcome the subjectivity associated with result determination of the fluorescence assay and plaque assay based methods. The novel ELISA based titration method can also be applied to high throughput detection.