Point-of-care testing for Toxoplasma gondii IgG/IgM using Toxoplasma ICT IgG-IgM test with sera from the United States and implications for developing countries.

Science.gov (United States)

Begeman, Ian J; Lykins, Joseph; Zhou, Ying; Lai, Bo Shiun; Levigne, Pauline; El Bissati, Kamal; Boyer, Kenneth; Withers, Shawn; Clouser, Fatima; Noble, A Gwendolyn; Rabiah, Peter; Swisher, Charles N; Heydemann, Peter T; Contopoulos-Ioannidis, Despina G; Montoya, Jose G; Maldonado, Yvonne; Ramirez, Raymund; Press, Cindy; Stillwaggon, Eileen; Peyron, François; McLeod, Rima

2017-06-01

Congenital toxoplasmosis is a serious but preventable and treatable disease. Gestational screening facilitates early detection and treatment of primary acquisition. Thus, fetal infection can be promptly diagnosed and treated and outcomes can be improved. We tested 180 sera with the Toxoplasma ICT IgG-IgM point-of-care (POC) test. Sera were from 116 chronically infected persons (48 serotype II; 14 serotype I-III; 25 serotype I-IIIa; 28 serotype Atypical, haplogroup 12; 1 not typed). These represent strains of parasites infecting mothers of congenitally infected children in the U.S. 51 seronegative samples and 13 samples from recently infected persons known to be IgG/IgM positive within the prior 2.7 months also were tested. Interpretation was confirmed by two blinded observers. A comparison of costs for POC vs. commercial laboratory testing methods was performed. We found that this new Toxoplasma ICT IgG-IgM POC test was highly sensitive (100%) and specific (100%) for distinguishing IgG/IgM-positive from negative sera. Use of such reliable POC tests can be cost-saving and benefit patients. Our work demonstrates that the Toxoplasma ICT IgG-IgM test can function reliably as a point-of-care test to diagnose Toxoplasma gondii infection in the U.S. This provides an opportunity to improve maternal-fetal care by using approaches, diagnostic tools, and medicines already available. This infection has serious, lifelong consequences for infected persons and their families. From the present study, it appears a simple, low-cost POC test is now available to help prevent morbidity/disability, decrease cost, and make gestational screening feasible. It also offers new options for improved prenatal care in low- and middle-income countries.

Sera of patients with celiac disease and neurologic disorders evoke a mitochondrial-dependent apoptosis in vitro.

Science.gov (United States)

Cervio, Elisabetta; Volta, Umberto; Verri, Manuela; Boschi, Federica; Pastoris, Ornella; Granito, Alessandro; Barbara, Giovanni; Parisi, Claudia; Felicani, Cristina; Tonini, Marcello; De Giorgio, Roberto

2007-07-01

The mechanisms underlying neurologic impairment in celiac disease remain unknown. We tested whether antineuronal antibody-positive sera of patients with celiac disease evoke neurodegeneration via apoptosis in vitro. SH-Sy5Y cells were exposed to crude sera, isolated immunoglobulin (Ig) G and IgG-depleted sera of patients with and without celiac disease with and without neurologic disorders, and antineuronal antibodies. Adsorption studies with gliadin and tissue transglutaminase (tTG) were performed in celiac disease sera. Apoptosis activated caspase-3, apaf-1, Bax, cytochrome c, cleaved caspase-8 and caspase-9 and mitochondrial respiratory chain complexes were evaluated with different methods. SH-Sy5Y cells exposed to antineuronal antibody-positive sera and isolated IgG from the same sera exhibited a greater percentage of TUNEL-positive nuclei than that of antineuronal antibody-negative sera. Neuroblasts exposed to antineuronal antibody-negative celiac disease sera also showed greater TUNEL positivity and apaf-1 immunolabeled cells than controls. Antigliadin- and anti-tTG-depleted celiac disease sera had an apoptotic effect similar to controls. Anti-caspase-3 immunostained cells were greater than controls when exposed to positive sera. The mitochondrial respiratory chain complex was reduced by positive sera. Western blot demonstrated only caspase-9 cleavage in positive sera. Cytochrome c and Bax showed reciprocal translocation (from mitochondria to cytoplasm and vice versa) after treatment with positive sera. Antineuronal antibodies and, to a lower extent, combined antigliadin and anti-tTG antibodies in celiac disease sera contribute to neurologic impairment via apoptosis. Apaf-1 activation with Bax and cytochrome c translocation suggest a mitochondrial-dependent apoptosis.

The Plasmodium falciparum pseudoprotease SERA5 regulates the kinetics and efficiency of malaria parasite egress from host erythrocytes.

Directory of Open Access Journals (Sweden)

Christine R Collins

2017-07-01

Full Text Available Egress of the malaria parasite Plasmodium falciparum from its host red blood cell is a rapid, highly regulated event that is essential for maintenance and completion of the parasite life cycle. Egress is protease-dependent and is temporally associated with extensive proteolytic modification of parasite proteins, including a family of papain-like proteins called SERA that are expressed in the parasite parasitophorous vacuole. Previous work has shown that the most abundant SERA, SERA5, plays an important but non-enzymatic role in asexual blood stages. SERA5 is extensively proteolytically processed by a parasite serine protease called SUB1 as well as an unidentified cysteine protease just prior to egress. However, neither the function of SERA5 nor the role of its processing is known. Here we show that conditional disruption of the SERA5 gene, or of both the SERA5 and related SERA4 genes simultaneously, results in a dramatic egress and replication defect characterised by premature host cell rupture and the failure of daughter merozoites to efficiently disseminate, instead being transiently retained within residual bounding membranes. SERA5 is not required for poration (permeabilization or vesiculation of the host cell membrane at egress, but the premature rupture phenotype requires the activity of a parasite or host cell cysteine protease. Complementation of SERA5 null parasites by ectopic expression of wild-type SERA5 reversed the egress defect, whereas expression of a SERA5 mutant refractory to processing failed to rescue the phenotype. Our findings implicate SERA5 as an important regulator of the kinetics and efficiency of egress and suggest that proteolytic modification is required for SERA5 function. In addition, our study reveals that efficient egress requires tight control of the timing of membrane rupture.

[Evaluation of angiogenic activity in sera from patients with interstitial lung diseases].

Science.gov (United States)

Zielonka, T M; Demkow, U; Kowalski, J; Kuś, J; Krychniak-Soszka, A; Radzikowska, E; Skopińska-Rózewska, E; Rowińska-Zakrzewska, E

1997-01-01

Angiogenesis is a process of new blood vessels' formation occurring in many physiological and pathological conditions. Neovascularisation is the principal vascular response in chronic inflammation and concomitant fibrotic process. Microvascular changes in various organ sites in sarcoidosis (BBS) and some of the symptoms of the disease may be related to microangiopathy. Moreover, vascular alterations were also observed in lung specimens from idiopathic pulmonary fibrosis (IPF) and avian fanciers lung (AFL) patients. The present study was aimed at testing the effects of serum from 43 patients with ILD (24 BBS, 8 AFL, 8 IPF, 3 DIPF--drug induced pulmonary fibrosis) and 11 healthy controls on angiogenic capability of normal blood peripheral mononuclear cells (PBMC) in the murine intradermal angiogenesis assay (according to Sidky and Auerbach). The data demonstrated that sera from ILD patients significantly enhanced angiogenic capacity of normal PBMC as compared to control sera (p < 0.001). The effect was more pronounced for AFL patients than for BBS and IPF ones (p < 0.05). Sera from DIPF did not stimulate angiogenesis compared to control sera. The data showed that sera from ILD patients constitute sources of mediators participating in angiogenesis. This phenomenon may play role in pathogenesis of chronic immunological processes in lung.

Discordant results obtained on testing sera from immunocompromised patients for cytomegalovirus IgG by enzyme-linked immunosorbent assay and radioimmunoassay

Energy Technology Data Exchange (ETDEWEB)

Booth, J C; El Mohandes, L; Tryhorn, Y S [London University (United Kingdom). St. George' s Hospital Medical School, Department of Medical Microbiology; Kangro, H O; Liu, K M [Saint Bartholomew' s Hospital, London (UK)

1989-10-01

The investigation of sera from immunocompromised patients for antibody to CMV by ELISA, RIA, immunofluorescence (IF) and complement-fixation (CF) revealed discrepancies that reflected differences in test specificity rather than sensitivity and suggested that for the long-term serological follow-up of such patients it would be advisable not to rely on only a single assay procedure. (author). 12 refs.; 8 figs.; 1 tab.

Reactivity of some mammalian sera with the bovine leukaemia virus env gene polypeptide expressed in Escherichia coli

International Nuclear Information System (INIS)

Slavikova, K.; Zajac, V.

1989-01-01

Sera from bovine leukaemia virus (BLV)-infected cattle and sheep were tested by radioimmunoassay and Western blot for their reactivity with 60,000 protein coded by the env gene of BLV and expressed in Escherichia coli. This protein, antigenically similar to BLV protein, reacted with antibodies against BLV antigens in the sera tested. (author). 3 figs., 1 tab., 13 refs

Complement-mediated killing of Borrelia burgdorferi by nonimmune sera from sika deer.

Science.gov (United States)

Nelson, D R; Rooney, S; Miller, N J; Mather, T N

2000-12-01

Various species of cervid deer are the preferred hosts for adult, black-legged ticks (Ixodes scapularis and Ixodes pacificus) in the United States. Although frequently exposed to the agent of Lyme disease (Borrelia burgdorferi), these animals, for the most part, are incompetent as transmission reservoirs. We examined the borreliacidal activity of normal and B. burgdorferi-immune sera from sika deer (Cervus nippon) maintained in a laboratory setting and compared it to that of similar sera from reservoir-competent mice and rabbits. All normal deer sera (NDS) tested killed > 90% of B. burgdorferi cells. In contrast, normal mouse and rabbit sera killed feeding exhibited IFA titers of 1:256, whereas sera from mice and rabbits similarly exposed had titers of > 1:1,024. Heat treatment (56 C, 30 min) of NDS reduced borreliacidal activity, with complement-mediated killing. The chelators EGTA and EDTA were used to block the classical or both the classical and alternative complement pathways, respectively. Addition of 10 mM EGTA to NDS had a negligible effect on borreliacidal activity, with > 90% of the cells killed. Addition of 10 mM EDTA reduced the killing to approximately 30%, whereas the addition of Mg2+ (10 mM) restored borreliacidal activity to NDS. The addition of zymosan A, an activator of the alternative pathway, increased the survival of B. burgdorferi cells to approximately 80% in NDS. These data suggest that the alternative complement activation pathway plays a major role in the borreliacidal activity of NDS. Additionally, 10 mM EGTA had almost no effect on the killing activity of B. burgdorferi-exposed deer sera, suggesting that the classical pathway is not involved in Borrelia killing, even in sera from B. burgdorferi-exposed deer.

Detection of immune complexes in sera of dogs with rheumatic and neoplastic diseases by 125I-Clq binding test

International Nuclear Information System (INIS)

Terman, D.S.; Moore, D.; Collins, J.; Johnston, B.; Person, D.; Templeton, J.; Poser, R.; Quinby, F.

1979-01-01

Some canine rheumatic and neoplastic diseases bear a striking clinical and serological resemblance to their counterparts in man. In the present study, human 125 I-Clq was employed in a radioimmunoassay for detection of immune complexes in sera of normal dogs and those with rheumatic and neoplastic diseases. Human 125 I-Clq showed binding of 16.7 +- 5.73% in a group of normal dog sera with binding of 32.5 +- 17.3% and 43.0 +- 16.0% in sera of dogs with rheumatic and neoplastic diseases. respectively. Human 125 I-Clq bound similar quantities of heat-aggregated canine and human gamma-globulin over a broad range of concentrations and human 125 I-Clq binding in canine sera was effectively inhibited by similar quantities of heat aggregated canine and human gamma-globulin. Seven of 12 dogs with elevated levels of Clq binding had active clinical and serological rheumatic disease (SLE or rheumatoid arthritis), while none of 7 dogs with values within the normal range had active clinical disease. All 5 dogs with widespread osteogenic sarcoma and all 4 dogs with high grade adenocarcinoma of the mammary gland had elevated Clq binding values while 2 animals with low grade malignancies without evident metastases did not. Thus, it appears that human 125 I-Clq may be employed to assay immune complexes in canine sera and may be a valuable technique for the study of dogs with various rheumatic and neoplastic diseases. (author)

Detection of anti-aquaporin-4 autoantibodies in the sera of Chinese neuromyelitis optica patients

Institute of Scientific and Technical Information of China (English)

Miao Li; Weiheng Su; Jie Wang; Francesco Pisani; Antonio Frigeri; Tonghui Ma

2013-01-01

In this study, we recruited 10 neuromyelitis optica patients, two multiple sclerosis patients and two myelitis patients. Chinese hamster lung fibroblast (V79) cells transfected with a human aquaporin-4-mCherry fusion protein gene were used to detect anti-aquaporin-4 antibody in neuromyelitis optica patient sera by immunofluorescence. Anti-aquaporin-4 autoantibody was stably detected by immunofluorescence in neuromyelitis optica patient sera exclusively. The sensitivity of the assay for neuromyelitis optica was 90% and the specificity for neuromyelitis optica was 100%. The anti-aquaporin-4 antibody titers in sera were tested with serial dilutions until the signal disappeared. A positive correlation was detected between Expanded Disability Status Scale scores and serum anti-aquaporin-4 antibody titers. The anti-aquaporin-4 antibody assay is highly sensitive and specific in the sera of Chinese neuromyelitis optica patients. Detection of aquaporin-4 autoantibody is important for the diagnosis and treatment of neuromyelitis optica.

Detection of anti-aquaporin-4 autoantibodies in the sera of Chinese neuromyelitis optica patients.

Science.gov (United States)

Li, Miao; Su, Weiheng; Wang, Jie; Pisani, Francesco; Frigeri, Antonio; Ma, Tonghui

2013-03-15

In this study, we recruited 10 neuromyelitis optica patients, two multiple sclerosis patients and two myelitis patients. Chinese hamster lung fibroblast (V79) cells transfected with a human aquaporin-4-mCherry fusion protein gene were used to detect anti-aquaporin-4 antibody in neuromyelitis optica patient sera by immunofluorescence. Anti-aquaporin-4 autoantibody was stably detected by immunofluorescence in neuromyelitis optica patient sera exclusively. The sensitivity of the assay for neuromyelitis optica was 90% and the specificity for neuromyelitis optica was 100%. The anti-aquaporin-4 antibody titers in sera were tested with serial dilutions until the signal disappeared. A positive correlation was detected between Expanded Disability Status Scale scores and serum anti-aquaporin-4 antibody titers. The anti-aquaporin-4 antibody assay is highly sensitive and specific in the sera of Chinese neuromyelitis optica patients. Detection of aquaporin-4 autoantibody is important for the diagnosis and treatment of neuromyelitis optica.

Detection of anti-aquaporin-4 autoantibodies in the sera of Chinese neuromyelitis optica patients★

Science.gov (United States)

Li, Miao; Su, Weiheng; Wang, Jie; Pisani, Francesco; Frigeri, Antonio; Ma, Tonghui

2013-01-01

In this study, we recruited 10 neuromyelitis optica patients, two multiple sclerosis patients and two myelitis patients. Chinese hamster lung fibroblast (V79) cells transfected with a human aquaporin-4-mCherry fusion protein gene were used to detect anti-aquaporin-4 antibody in neuromyelitis optica patient sera by immunofluorescence. Anti-aquaporin-4 autoantibody was stably detected by immunofluorescence in neuromyelitis optica patient sera exclusively. The sensitivity of the assay for neuromyelitis optica was 90% and the specificity for neuromyelitis optica was 100%. The anti-aquaporin-4 antibody titers in sera were tested with serial dilutions until the signal disappeared. A positive correlation was detected between Expanded Disability Status Scale scores and serum anti-aquaporin-4 antibody titers. The anti-aquaporin-4 antibody assay is highly sensitive and specific in the sera of Chinese neuromyelitis optica patients. Detection of aquaporin-4 autoantibody is important for the diagnosis and treatment of neuromyelitis optica. PMID:25206717

Long-term sera storage does not significantly modify the interpretation of toxoplasmosis serologies.

Science.gov (United States)

Dard, C; Bailly, S; Drouet, T; Fricker-Hidalgo, H; Brenier-Pinchart, M P; Pelloux, H

2017-03-01

Serological investigation of Toxoplasma gondii can answer many questions about toxoplasmosis in human pathology. Along these lines, studies on serum storage in biobanks need to be performed especially in terms of determining the impact of storage on relevance of sera analysis after freezing. This study assessed the impact of long-term sera storage on the stability of anti-Toxoplasma immunoglobulins. The stability of anti-Toxoplasma IgG and IgM was studied in 244 and 242 sera respectively, stored at -20°C from one month to ten years. ELISA-immunoassay (Vidas®, bioMérieux) was used for initial and post-storage analyses. Linear models for repeated measures and subgroup analyses were performed to assess the effect of storage duration and sample characteristics on immunoglobulins stability. Until ten years, the variability attributed to storage (maximum 8.07% for IgG, 13.17% for IgM) was below the variations inherent to the serological technique and allowed by quality assurance systems (15%). Subgroup analysis reported no variation attributed to sera storage. Serological interpretation was modified for 3 sera (1.2%) tested for IgM, all stored more than seven years. Anti-Toxoplasma immunoglobulins can reliably be measured for at least up to six years of storage with no modification of interpretation of toxoplasmosis serologies. Copyright © 2017 Elsevier B.V. All rights reserved.

Ausab test - a radioimmunoassay in the solid phase for the detection of antibodies against HBsAg

Energy Technology Data Exchange (ETDEWEB)

Lange, W; Koehler, H; Apodaca, J [Bundesgesundheitsamt, Berlin (Germany, F.R.). Abt. fuer Virologie

1974-01-01

The suitability of a newly developed RIA (Ausab test, Abbott laboratories) for the detection of antibodies against the Hepatitis B (surface) antigen (HBsAg) has been tested on 1968 sera of blood donors. Reproducibility test and specificity test were included. During the examination the technique of the Ausab test was changed from the original one-step to the two-step procedure. With the one-step technique 431 of 1968 sera (21.9%) showed positive reactions. During the test problems concerning the differentiation between positive and negative results were encountered due to high unspecific binding of radioactivity. Positive sera were re-examined employing the two-step technique and only 77.4% of the previously positive reactions could be reproduced. 50 sera that were negative in the one-step test remained negative in the two-step test. Difficulties concerning nonspecific binding of radioactivity as observed with the one-step technique were not encountered with the two-step procedure. On 92 of the confirmed positive sera a specificity test was run and 90 (97.8%) proved to be specific. A calculated 16.5% of the blood donors in Berlin have to be considered as antibody carriers. Only 6 of the Ausab positive sera were found to give positive reactions in CEP. 3 sera that were initially positive in CEP could neither be confirmed in a repeated CEP nor in the Ausab test.

The Ausab test - a radioimmunoassay in the solid phase for the detection of antibodies against HBsAg

International Nuclear Information System (INIS)

Lange, W.; Koehler, H.; Apodaca, J.

1974-01-01

The suitability of a newly developed RIA (Ausab test, Abbott laboratories) for the detection of antibodies against the Hepatitis B (surface) antigen (HBsAg) has been tested on 1968 sera of blood donors. Reproducibility test and specificity test were included. During the examination the technique of the Ausab test was changed from the original one-step to the two-step procedure. With the one-step technique 431 of 1968 sera (21.9%) showed positive reactions. During the test problems concerning the differentiation between positive and negative results were encountered due to high unspecific binding of radioactivity. Positive sera were re-examined employing the two-step technique and only 77.4% of the previously positive reactions could be reproduced. 50 sera that were negative in the one-step test remained negative in the two-step test. Difficulties concerning nonspecific binding of radioactivity as observed with the one-step technique were not encountered with the two-step procedure. On 92 of the confirmed positive sera a specificity test was run and 90 (97.8%) proved to be specific. A calculated 16.5% of the blood donors in Berlin have to be considered as antibody carriers. Only 6 of the Ausab positive sera were found to give positive reactions in CEP. 3 sera that were initially positive in CEP could neither be confirmed in a repeated CEP nor in the Ausab test. (orig.) [de

Effect of animal sera on Bacillus anthracis Sterne spore germination and vegetative cell growth.

Science.gov (United States)

Bensman, M D; Mackie, R S; Minter, Z A; Gutting, B W

2012-08-01

 The aims of this work were to investigate the effects of sera on B. anthracis Sterne germination and growth. Sera examined included human, monkey and rabbit sera, as well as sera from eight other species.  Standard dilution plate assay (with and without heat kill) was used as a measure of germination, and spectroscopy was used to measure growth. In addition, a Coulter Counter particle counter was used to monitor germination and growth based on bacterial size. Spores germinated best in foetal bovine and monkey sera, moderately with human sera and showed limited germination in the presence of rabbit or rat sera. Vegetative bacteria grew best in foetal bovine sera and moderately in rabbit sera. Human and monkey sera supported little growth of vegetative bacteria.  The data suggested sera can have a significant impact on germination and growth of Sterne bacteria.  These data should be considered when conducting in vitro cell culture studies and may aid in interpreting in vivo infection studies. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

Characterization of human septic sera induced gene expression modulation in human myocytes

Science.gov (United States)

Hussein, Shaimaa; Michael, Paul; Brabant, Danielle; Omri, Abdelwahab; Narain, Ravin; Passi, Kalpdrum; Ramana, Chilakamarti V.; Parrillo, Joseph E.; Kumar, Anand; Parissenti, Amadeo; Kumar, Aseem

2009-01-01

To gain a better understanding of the gene expression changes that occurs during sepsis, we have performed a cDNA microarray study utilizing a tissue culture model that mimics human sepsis. This study utilized an in vitro model of cultured human fetal cardiac myocytes treated with 10% sera from septic patients or 10% sera from healthy volunteers. A 1700 cDNA expression microarray was used to compare the transcription profile from human cardiac myocytes treated with septic sera vs normal sera. Septic sera treatment of myocytes resulted in the down-regulation of 178 genes and the up-regulation of 4 genes. Our data indicate that septic sera induced cell cycle, metabolic, transcription factor and apoptotic gene expression changes in human myocytes. Identification and characterization of gene expression changes that occur during sepsis may lead to the development of novel therapeutics and diagnostics. PMID:19684886

The Ausab test - a radioimmunoassay in the solid phase for the detection of antibodies against HBsAg

International Nuclear Information System (INIS)

Lange, W.; Koehler, H.; Apodaca, J.

1974-01-01

The suitability of a newly developed RIA (Ausab test, Abbott Laboratories) for the detection of antibodies against the Hepatitis B (surface) antigen (HBsAg) has been tested on 1,968 sera of blood donors. Reproducibility test and specificity test were included. During the examination the technique of the Ausab test was changed from the original one-step to the two-step procedure. With the one-step technique 431 of 1,968 sera (21.9%) showed positive reactions. During the test problems concerning the differentiation between positive and negative results were encountered due to high unspecific binding of radioactivity. Positive sera were reexamined employing the two-step technique and only 77.4% of the previously positive reactions could be reproduced. 50 sera that were negative in the one-step test remained negative in the two-step test. Difficulties concerning nonspecific binding of radioactivity as observed with the one-step technique were not ecountered with the two-step procedure. On 92 of the confirmed positive sera a specificity test was run and 90 (97.8%) proved to be specific. A calculated 16.5% of the blood donors in Berlin have to be considered as antibody carriers. Only 6 of the Ausab positive sera were found to give positive reactions in CEP. 3 sera that were initially positive in CEP could neither be confirmed in a repeated CEP nor in the Ausab test. (orig.) [de

Characterization of human septic sera induced gene expression modulation in human myocytes

OpenAIRE

Hussein, Shaimaa; Michael, Paul; Brabant, Danielle; Omri, Abdelwahab; Narain, Ravin; Passi, Kalpdrum; Ramana, Chilakamarti V.; Parrillo, Joseph E.; Kumar, Anand; Parissenti, Amadeo; Kumar, Aseem

2009-01-01

To gain a better understanding of the gene expression changes that occurs during sepsis, we have performed a cDNA microarray study utilizing a tissue culture model that mimics human sepsis. This study utilized an in vitro model of cultured human fetal cardiac myocytes treated with 10% sera from septic patients or 10% sera from healthy volunteers. A 1700 cDNA expression microarray was used to compare the transcription profile from human cardiac myocytes treated with septic sera vs normal sera....

Trypanosoma brucei gambiense adaptation to different mammalian sera is associated with VSG expression site plasticity.

Science.gov (United States)

Cordon-Obras, Carlos; Cano, Jorge; González-Pacanowska, Dolores; Benito, Agustin; Navarro, Miguel; Bart, Jean-Mathieu

2013-01-01

Trypanosoma brucei gambiense infection is widely considered an anthroponosis, although it has also been found in wild and domestic animals. Thus, fauna could act as reservoir, constraining the elimination of the parasite in hypo-endemic foci. To better understand the possible maintenance of T. b. gambiense in local fauna and investigate the molecular mechanisms underlying adaptation, we generated adapted cells lines (ACLs) by in vitro culture of the parasites in different mammalian sera. Using specific antibodies against the Variant Surface Glycoproteins (VSGs) we found that serum ACLs exhibited different VSG variants when maintained in pig, goat or human sera. Although newly detected VSGs were independent of the sera used, the consistent appearance of different VSGs suggested remodelling of the co-transcribed genes at the telomeric Expression Site (VSG-ES). Thus, Expression Site Associated Genes (ESAGs) sequences were analysed to investigate possible polymorphism selection. ESAGs 6 and 7 genotypes, encoding the transferrin receptor (TfR), expressed in different ACLs were characterised. In addition, we quantified the ESAG6/7 mRNA levels and analysed transferrin (Tf) uptake. Interestingly, the best growth occurred in pig and human serum ACLs, which consistently exhibited a predominant ESAG7 genotype and higher Tf uptake than those obtained in calf and goat sera. We also detected an apparent selection of specific ESAG3 genotypes in the pig and human serum ACLs, suggesting that other ESAGs could be involved in the host adaptation processes. Altogether, these results suggest a model whereby VSG-ES remodelling allows the parasite to express a specific set of ESAGs to provide selective advantages in different hosts. Finally, pig serum ACLs display phenotypic adaptation parameters closely related to human serum ACLs but distinct to parasites grown in calf and goat sera. These results suggest a better suitability of swine to maintain T. b. gambiense infection supporting

Determination of low tetanus or diphtheria antitoxin titers in sera by a toxin neutralization assay and a modified toxin-binding inhibition test

Directory of Open Access Journals (Sweden)

M.H. Sonobe

2007-01-01

Full Text Available A method for the screening of tetanus and diphtheria antibodies in serum using anatoxin (inactivated toxin instead of toxin was developed as an alternative to the in vivo toxin neutralization assay based on the toxin-binding inhibition test (TOBI test. In this study, the serum titers (values between 1.0 and 19.5 IU measured by a modified TOBI test (Modi-TOBI test and toxin neutralization assays were correlated (P < 0.0001. Titers of tetanus or diphtheria antibodies were evaluated in serum samples from guinea pigs immunized with tetanus toxoid, diphtheria-tetanus or triple vaccine. For the Modi-TOBI test, after blocking the microtiter plates, standard tetanus or diphtheria antitoxin and different concentrations of guinea pig sera were incubated with the respective anatoxin. Twelve hours later, these samples were transferred to a plate previously coated with tetanus or diphtheria antitoxin to bind the remaining anatoxin. The anatoxin was then detected using a peroxidase-labeled tetanus or diphtheria antitoxin. Serum titers were calculated using a linear regression plot of the results for the corresponding standard antitoxin. For the toxin neutralization assay, L+/10/50 doses of either toxin combined with different concentrations of serum samples were inoculated into mice for anti-tetanus detection, or in guinea pigs for anti-diphtheria detection. Both assays were suitable for determining wide ranges of antitoxin levels. The linear regression plots showed high correlation coefficients for tetanus (r² = 0.95, P < 0.0001 and for diphtheria (r² = 0.93, P < 0.0001 between the in vitro and the in vivo assays. The standardized method is appropriate for evaluating titers of neutralizing antibodies, thus permitting the in vitro control of serum antitoxin levels.

Effects of amyotrophic lateral sclerosis sera on cultured cholinergic neurons

International Nuclear Information System (INIS)

Touzeau, G.; Kato, A.C.

1983-01-01

Dissociated monolayer cultures of chick ciliary ganglion neurons have been used to study the effects of control and ALS sera. The cultured neurons survive and extend neurites for a minimum of 2 weeks in a standard tissue culture medium that contains 10% heat-inactivated human serum. Three parameters of the neurons have been examined when cultured in control and ALS sera for 8 to 12 days: (1) neuronal survival, (2) activity of the enzyme choline acetyltransferase, and (3) synthesis of 3 H-acetylcholine using 3 H-choline as precursor. ALS sera cause a small decrease in these three parameters, but this difference is not significant

Local production of donkey anti-rabbit's sera for human prolactin radioimmunoassay

International Nuclear Information System (INIS)

Hassan, Ammar Mohamed Elamin

2001-11-01

Pure Rabbit's IgG was used in this study to raise donkey anti rabbit's sera to be used as separating agent in radioimmunoassay. Two healthy donkeys have been immunized. The anti rabbit's sera have been titrated as (i) crude, (ii) purified and dialysed coupled to magnetic particles. Then this antibody was used as separating agent in a radioimmunoassay for measurement of human prolactin (PRL). Coupled Sudanese donkey and rabbit's sera (Sud-DARS) was used as 1/8 titre using chelsea RIA kit for human prolactin while 1/200 of liquid Sud-DARS was found to be the best titre using the Chinese kit. The best condition for estimation of the prolactin were optimized by determining the suitable incubation time and temperature. The assay can be done at room temperature but it should be incubated for 6 hours as recommended by the Chinese kit. Validity tests were done. The regression coefficients were 0.994 and 0.999 for linearity and recovery tests respectively. Measurement of human PRL wa found to be reproducible using Sud-DARS as separating agent since the coefficient of variation (C.V. %) was found to be less than 15% for both within batch and between assays. Comparing Sud D ARS to the Chinese kit, separating agent as reference agent, regression coefficient was found to be 0.977 which indicate that Sud-DARS can be used as separating agent. Prolactin in Sudanese subject was determined using the Chinese kit the Sud-DARS as separating agent. The ranges were 74-398 mIU/L in males and 102-414 mI/L in the preovulatory phase for the females while in the post ovulatory phase it was 114-442 mIU/L. Ovulation was confirmed by measurement of progesterone level 7 days before the next suspected mensuration. (Author)

Modification of the Rappaport rapid test in large-scale testing for syphilis. Evaluation of the rapid plate and rapid card tests.

Science.gov (United States)

Ghinsberg, R; Meir, E; Blumstein, G; Kafeman, R

1975-11-01

The Rappaport rapid (RR) plate and card tests were developed as modifications of the RR tube test to permit rapid and inexpensive screening of large numbers of subjects for the diagnosis of syphilis. More than 2,000 sera were examined in parallel by the Venereal Disease Research Laboratory (VDRL) slide test, the rapid plasma reagin (RPR) card test and the RR plate and card tests. There was complete agreement between the RR plate and card tests and the VDRL slide and RPR card tests in 96.6% of sera. In a selected group of 1,530 sera examined, in addition, by the fluorescent treponemal antibody absorption (FTA-ABS) test, there was agreement between the RR plate and card tests and the FTA-ABS test in 74.3% of sera and between the VDRL and RPR tests and the FTA-ABS test in 73.7% of sera. The RR plate test was found to be sufficiently sensitive and specific for the diagnosis of syphilis, although the VDRL slide test is perhaps more sensitive in primary and late latent syphilis. Since the antigen used in the RR tests is colored and stable and the sera do not require inactivation before the test, the tests are easier to perform than the VDRL slide test: the RR plate and card tests could therefore replace the VDRL test as a screening test, with hardly any loss of accuracy.

Effects of proposed adipogenic factors in fetal swine sera upon preadipocyte development

International Nuclear Information System (INIS)

Ramsay, T.G.; Hausman, G.J.; Martin, R.J.

1986-01-01

Genetic obesity has been detected in fetal pigs which suggests primary factors that cause the obesity develop prenatally. Growth hormone and thyroid hormones have been implicated as regulatory factors in fetal serum for preadipocyte differentiation. This experiment examined effects of growth hormone (GH) and thyroxine (T4) addition upon preadipocyte proliferation and differentiation when supplemented to deficient fetal pig sea. Hormones were added to decapitated fetal pig (Decap) sera to concentrations present in intact littermate (Reference) sera. Primary stromal-vascular cell cultures were prepared from rat inguinal adipose tissue. Cells were incubated with 5% decap or reference sera and hormones in media 199 during: days 1 to 5 for a 3 H-thymidine incorporation assay; days 1 to 15 for assay of α-glycerol phosphate dehydrogenase; days 5 to 14 for a complete differentiation assay. Decap sera promoted less proliferation and enzyme differentiation than reference sera with no effect of GH addition. GH reduced detection of lipid accumulating cells on percol density gradients by 81%. T4 addition stimulated preadipocyte multiplication and produced a 30% increase in completely differentiated preadipocytes. These results indicate thyroid hormones are important components of fetal sera for regulation of preadipocyte development, whereas GH may only affect cellular metabolism

Protein array profiling of tic patient sera reveals a broad range and enhanced immune response against Group A Streptococcus antigens.

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Mauro Bombaci

Full Text Available The human pathogen Group A Streptococcus (Streptococcus pyogenes, GAS is widely recognized as a major cause of common pharyngitis as well as of severe invasive diseases and non-suppurative sequelae associated with the existence of GAS antigens eliciting host autoantibodies. It has been proposed that a subset of paediatric disorders characterized by tics and obsessive-compulsive symptoms would exacerbate in association with relapses of GAS-associated pharyngitis. This hypothesis is however still controversial. In the attempt to shed light on the contribution of GAS infections to the onset of neuropsychiatric or behavioral disorders affecting as many as 3% of children and adolescents, we tested the antibody response of tic patient sera to a representative panel of GAS antigens. In particular, 102 recombinant proteins were spotted on nitrocellulose-coated glass slides and probed against 61 sera collected from young patients with typical tic neuropsychiatric symptoms but with no overt GAS infection. Sera from 35 children with neither tic disorder nor overt GAS infection were also analyzed. The protein recognition patterns of these two sera groups were compared with those obtained using 239 sera from children with GAS-associated pharyngitis. This comparative analysis identified 25 antigens recognized by sera of the three patient groups and 21 antigens recognized by tic and pharyngitis sera, but poorly or not recognized by sera from children without tic. Interestingly, these antigens appeared to be, in quantitative terms, more immunogenic in tic than in pharyngitis patients. Additionally, a third group of antigens appeared to be preferentially and specifically recognized by tic sera. These findings provide the first evidence that tic patient sera exhibit immunological profiles typical of individuals who elicited a broad, specific and strong immune response against GAS. This may be relevant in the context of one of the hypothesis proposing that GAS

BACTERICIDAL ACTIVITY OF HUMAN SERA AGAINST ...

African Journals Online (AJOL)

hi-tech

East African Medical Journal Vol. 77 No. 12 December 2000. BACTERICIDAL ACTIVITY OF HUMAN SERA AGAINST SALMONELLA TYPHI AND SALMONELLA PARATYPHI A, B, C. E.O. Igumbor, BSc, MSc, PhD, Department of Medical Microbiology, School of Medicine, University of Zimbabwe P.O. Box Al78, Avondale, ...

Sera from children with autism induce autistic features which can be rescued with a CNTF small peptide mimetic in rats.

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Kazim, Syed Faraz; Cardenas-Aguayo, Maria Del Carmen; Arif, Mohammad; Blanchard, Julie; Fayyaz, Fatima; Grundke-Iqbal, Inge; Iqbal, Khalid

2015-01-01

Autism is a neurodevelopmental disorder characterized clinically by impairments in social interaction and verbal and non-verbal communication skills as well as restricted interests and repetitive behavior. It has been hypothesized that altered brain environment including an imbalance in neurotrophic support during early development contributes to the pathophysiology of autism. Here we report that sera from children with autism which exhibited abnormal levels of various neurotrophic factors induced cell death and oxidative stress in mouse primary cultured cortical neurons. The effects of sera from autistic children were rescued by pre-treatment with a ciliary neurotrophic factor (CNTF) small peptide mimetic, Peptide 6 (P6), which was previously shown to exert its neuroprotective effect by modulating CNTF/JAK/STAT pathway and LIF signaling and by enhancing brain derived neurotrophic factor (BDNF) expression. Similar neurotoxic effects and neuroinflammation were observed in young Wistar rats injected intracerebroventricularly with autism sera within hours after birth. The autism sera injected rats demonstrated developmental delay and deficits in social communication, interaction, and novelty. Both the neurobiological changes and the behavioral autistic phenotype were ameliorated by P6 treatment. These findings implicate the involvement of neurotrophic imbalance during early brain development in the pathophysiology of autism and a proof of principle of P6 as a potential therapeutic strategy for autism.

Cytolytic tests with hyperimmune patient sera is a good prognostic tool in racotumomab immunotherapy in advanced non-small cell lung cancer

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Necdet Uskent

2017-10-01

Full Text Available Aberrant accumulation of a specific sialic acid has been shown to exist in many human malignant cell membranes termed as N-glycolyl neuraminic acid (NeuGc. This particular ganglioside do not normally exist in normal human cells, due to the lack of an enzyme (cytidine monophospho-N-acetyl-neuraminic acid which is responsible for the synthesis of N—glycolyl neurominic acid. The aberrant expression of NeuGcGM3 ganglioside in the cell surface of certain human tumors, made this molecule an attractive target for immunotherapy. By using 14 F7 monoclonal antibody directed to identify NeuGcGM3 in the tumor tissue, it is possible to select patients for anti-NeuGcGM3 immunotherapy. Racotumomab is an anti-idiotype vaccine, being a mirror image of NeuGcGm3 mimics this ganglioside and triggers an immune response. Antibodies reactive to NeuGcGM3 ganglioside in the vaccinated patient’s sera have cytotoxic anti-tumor properties which can be assessed in L1210 cell line, expressing this ganglioside.In this study, we monitored 12 patients with advanced non-small cell lung cancer (NSCLC who are on racotumomab vaccine maintenance following chemotherapy. Cytotoxic tests with vaccinated patients’ sera were performed using L1210 cell lines at the 3rd, 6th, 9th, and 12th months of vaccination and the results were compared with clinical outcomes. Serum antibodies to NeuGcGm3 ganglioside were also checked before initiation and thereafter with the same intervals. The aim of the study was to investigate the value of antibodies and cytotoxic test as biomarkers for treatment outcome. Our observation confirmed that consistently higher cytotoxicity rates in the cell culture correlated with better progression free survivals of the patients who are on racotumomab maintenance.

Antinuclear antibodies in the sera of patients with nasopharyngeal carcinoma

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Takimoto, T.; Ishikawa, S.; Masuda, K.; Tanaka, S.; Yoshizaki, T.; Umeda, R. (Kanazawa Univ. (Japan))

1989-11-01

We studied the production of heterophile antinuclear antibodies (ANAs) in the sera of 50 patients, 20 with nasopharyngeal carcinoma (NPC) and 30 with other head and neck cancers (laryngeal cancer and maxillary cancer), before and after radiation therapy. A higher incidence of ANAs was found in the sera of patients with NPC and ANA production in these patients was higher after radiation therapy. We therefore performed in vitro experiments to explore the mechanisms of ANA production in the serum of postirradiated NPC patients. X-ray-irradiated NPC-derived cells (NPC-KT) produced a large amount of Epstein-Barr virus (NPC EBV) compared with non-irradiated NPC-KT cells. Nasopharyngeal carcinoma EBV-infected lymphocytes produced high levels of ANAs. These data suggest that lymphocytes infected by EBV from NPC cells may produce ANAs in the sera of NPC patients.

Hemagglutination treponemal test for syphilis.

OpenAIRE

Friedly, G; Zartarian, M V; Wood, J C; Floyd, C M; Peterson, E M; de la Maza, L M

1983-01-01

Sera from 290 hospital patients were tested to compare the sensitivity, specificity, and reproducibility of the hemagglutination treponemal test for syphilis (HATTS) with the fluorescent treponemal antibody absorption test (FTA-ABS). Complete agreement was obtained between the methods when 142 syphilitic sera from patients with various stages of syphilis were tested. By using clinical histories, the specificity with 148 nonsyphilitic sera was determined to be 100% for the HATTS and 96.6% (143...

Detection of toxoplasma-specific immunoglobulin G in human sera: performance comparison of in house Dot-ELISA with ECLIA and ELISA.

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Teimouri, Aref; Modarressi, Mohammad Hossein; Shojaee, Saeedeh; Mohebali, Mehdi; Zouei, Nima; Rezaian, Mostafa; Keshavarz, Hossein

2018-05-08

In the current study, performance of electrochemiluminescence immunoassay (ECLIA) in detection of anti-toxoplasma IgG in human sera was compared with that of enzyme-linked immunosorbent assay (ELISA). Furthermore, performance of an in house Dot-ELISA in detection of anti-toxoplasma IgG was compared with that of ECLIA and ELISA. In total, 219 human sera were tested to detect anti-toxoplasma IgG using Dynex DS2® and Roche Cobas® e411 Automated Analyzers. Discordant results rechecked using immunofluorescence assay (IFA). Then, sera were used in an in house Dot-ELISA to assess toxoplasma-specific IgG. Of the 219 samples, two samples were found undetermined using ECLIA but reactive using ELISA. Using IFA, the two sera were reported unreactive. Furthermore, two samples were found reactive using ECLIA and unreactive using ELISA. These samples were reported reactive using IFA. The overall agreement for the two former methods was 98% (rZ0.98.1; P house Dot-ELISA included sensitivity of 79.5, specificity of 78.2, and accuracy of 78.9%, compared to ECLIA and ELISA. Positive and negative predictive values included 82.9 and 74.2%, respectively. A 100% sensitivity was found in in house Dot-ELISA for highly reactive sera in ECLIA and ELISA. ECLIA is appropriate for the first-line serological screening tests and can replace ELISA due to high speed, sensitivity, and specificity, particularly in large laboratories. Dot-ELISA is a rapid, sensitive, specific, cost-effective, user-friendly, and field-portable technique and hence can be used for screening toxoplasmosis, especially in rural fields or less equipped laboratories.

Sera from remitting and secondary progressive multiple sclerosis patients disrupt the blood-brain barrier.

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Shimizu, Fumitaka; Tasaki, Ayako; Sano, Yasuteru; Ju, Mihua; Nishihara, Hideaki; Oishi, Mariko; Koga, Michiaki; Kawai, Motoharu; Kanda, Takashi

2014-01-01

Pathological destruction of blood-brain barrier (BBB) has been thought to be the initial key event in the process of developing multiple sclerosis (MS). The purpose of the present study was to clarify the possible molecular mechanisms responsible for the malfunction of BBB by sera from relapse-remitting MS (RRMS) and secondary progressive MS (SPMS) patients. We evaluated the effects of sera from the patients in the relapse phase of RRMS (RRMS-R), stable phase of RRMS (RRMS-S) and SPMS on the expression of tight junction proteins and vascular cell adhesion protein-1 (VCAM-1), and on the transendothelial electrical resistance (TEER) in human brain microvascular endothelial cells (BMECs). Sera from the RRMS-R or SPMS patients decreased the claudin-5 protein expression and the TEER in BMECs. In RRMS-R, this effect was restored after adding an MMP inhibitor, and the MMP-2/9 secretion by BMECs was significantly increased after the application of patients' sera. In SPMS, the immunoglobulin G (IgG) purified from patients' sera also decreased the claudin-5 protein expression and the TEER in BMECs. The sera and purified IgG from all MS patients increased the VCAM-1 protein expression in BMECs. The up-regulation of autocrine MMP-2/9 by BMECs after exposure to sera from RRMS-R patients or the autoantibodies against BMECs from SPMS patients can compromise the BBB. Both RRMS-S and SPMS sera increased the VCAM-1 expression in the BBB, thus indicating that targeting the VCAM-1 in the BBB could represent a possible therapeutic strategy for even the stable phase of MS and SPMS.

Sera from remitting and secondary progressive multiple sclerosis patients disrupt the blood-brain barrier.

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Fumitaka Shimizu

Full Text Available BACKGROUND: Pathological destruction of blood-brain barrier (BBB has been thought to be the initial key event in the process of developing multiple sclerosis (MS. The purpose of the present study was to clarify the possible molecular mechanisms responsible for the malfunction of BBB by sera from relapse-remitting MS (RRMS and secondary progressive MS (SPMS patients. METHODS: We evaluated the effects of sera from the patients in the relapse phase of RRMS (RRMS-R, stable phase of RRMS (RRMS-S and SPMS on the expression of tight junction proteins and vascular cell adhesion protein-1 (VCAM-1, and on the transendothelial electrical resistance (TEER in human brain microvascular endothelial cells (BMECs. RESULTS: Sera from the RRMS-R or SPMS patients decreased the claudin-5 protein expression and the TEER in BMECs. In RRMS-R, this effect was restored after adding an MMP inhibitor, and the MMP-2/9 secretion by BMECs was significantly increased after the application of patients' sera. In SPMS, the immunoglobulin G (IgG purified from patients' sera also decreased the claudin-5 protein expression and the TEER in BMECs. The sera and purified IgG from all MS patients increased the VCAM-1 protein expression in BMECs. CONCLUSIONS: The up-regulation of autocrine MMP-2/9 by BMECs after exposure to sera from RRMS-R patients or the autoantibodies against BMECs from SPMS patients can compromise the BBB. Both RRMS-S and SPMS sera increased the VCAM-1 expression in the BBB, thus indicating that targeting the VCAM-1 in the BBB could represent a possible therapeutic strategy for even the stable phase of MS and SPMS.

Evaluation of commercial antiglobulin sera over a two-year period. Part I. Anti-beta 1A, anti-alpha 2D, and anti-beta 1E levels.

Science.gov (United States)

Issitt, P D; Issitt, C H; Wilkinson, S L

1974-01-01

Antiglobulin sera from nine different manufacturers have been tested, over a two-year period, for their ability to detect the complement components beta 1A, alpha 2D, and beta 1E. The results demontrate considerable variation in the abilities of sera from different manufacturers to detect these components and indicate that not all sera on the market are suitable reagents for diagnostic use and compatibility tests. The results also show that there is considerable variation between different lots of serum from some of the manufacturers. In general, the anti-complement levels of these reagents have increased during the two-year period of study but not all companies produce suitable reagents for routine use.

Antibody reaction of human anti-Toxoplasma gondii positive and negative sera with Neospora caninum antigens

OpenAIRE

Nam, Ho-Woo; Kang, Seung-Won; Choi, Won-Young

1998-01-01

Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7%) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG1) and 22 kD...

Sera from Children with Autism Induce Autistic Features Which Can Be Rescued with a CNTF Small Peptide Mimetic in Rats

Science.gov (United States)

Kazim, Syed Faraz; Cardenas-Aguayo, Maria del Carmen; Arif, Mohammad; Blanchard, Julie; Fayyaz, Fatima; Grundke-Iqbal, Inge; Iqbal, Khalid

2015-01-01

Autism is a neurodevelopmental disorder characterized clinically by impairments in social interaction and verbal and non-verbal communication skills as well as restricted interests and repetitive behavior. It has been hypothesized that altered brain environment including an imbalance in neurotrophic support during early development contributes to the pathophysiology of autism. Here we report that sera from children with autism which exhibited abnormal levels of various neurotrophic factors induced cell death and oxidative stress in mouse primary cultured cortical neurons. The effects of sera from autistic children were rescued by pre-treatment with a ciliary neurotrophic factor (CNTF) small peptide mimetic, Peptide 6 (P6), which was previously shown to exert its neuroprotective effect by modulating CNTF/JAK/STAT pathway and LIF signaling and by enhancing brain derived neurotrophic factor (BDNF) expression. Similar neurotoxic effects and neuroinflammation were observed in young Wistar rats injected intracerebroventricularly with autism sera within hours after birth. The autism sera injected rats demonstrated developmental delay and deficits in social communication, interaction, and novelty. Both the neurobiological changes and the behavioral autistic phenotype were ameliorated by P6 treatment. These findings implicate the involvement of neurotrophic imbalance during early brain development in the pathophysiology of autism and a proof of principle of P6 as a potential therapeutic strategy for autism. PMID:25769033

Food allergy: a clinician's criteria for including sera in a serum bank.

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Ballmer-Weber, B K; Fernández-Rivas, M

2008-10-01

Safety assessment for genetically-engineered crop plants includes assessment for allergic responses. To facilitate this assessment, serum banks should contain well-characterised sera from patients with confirmed food allergies. A serum is defined as well-characterised if it is taken from a patient who has a convincing history of allergic responses to a known allergen or an allergen-containing food, a positive skin prick test (or elevated IgE response), and a positive response in a clinical food challenge.

Analysis of Cl and Na in Hyperimmune Sera by NAA

International Nuclear Information System (INIS)

Baptista, T. S.; Zamboni, C. B.; Marcelino, J. R.

2011-01-01

The Cl and Na concentration values in four types of hyperimmune sera (anti-Bothrops, anti-Diphtheria, anti-Rabies and anti-Tetanus) used for immunological therapy were determined using Neutron Activation Analysis (NAA). These data were compatible with the specifications established by the Word Health Organization (WHO-OMS) and with the Brazilian Official Pharmacopea (Pharmaceutical Code Official of the Country). These data are an important support for quality control of hyperimmune sera production at Butantan Institute (Sao Paulo city, Brazil), responsible for supplying the Brazilian market.

Antibodies against glucan, chitin, and Saccharomyces cerevisiae mannan as new biomarkers of Candida albicans infection that complement tests based on C. albicans mannan.

Science.gov (United States)

Sendid, B; Dotan, N; Nseir, S; Savaux, C; Vandewalle, P; Standaert, A; Zerimech, F; Guery, B P; Dukler, A; Colombel, J F; Poulain, D

2008-12-01

Antibodies against Saccharomyces cerevisiae mannan (ASCA) and antibodies against synthetic disaccharide fragments of glucans (ALCA) and chitin (ACCA) are biomarkers of Crohn's disease (CD). We previously showed that Candida albicans infection generates ASCA. Here, we explored ALCA and ACCA as possible biomarkers of invasive C. albicans infection (ICI). ASCA, ALCA, ACCA, and Candida mannan antigen and antibody detection tests were performed on 69 sera obtained sequentially from 18 patients with ICIs proven by blood culture, 59 sera from CD patients, 47 sera from hospitalized subjects colonized by Candida species (CZ), and 131 sera from healthy controls (HC). ASCA, ALCA, and ACCA levels in CD and ICI patients were significantly different from those in CZ and HC subjects (PACCA, and Platelia Candida tests, 100% of ICIs were detected, with the kinetics of the antibody response depending on the patient during the time course of infection. A large number of sera presented with more than three positive tests. This is the first evidence that the detection of antibodies against chitin and glucans has diagnostic value in fungal infections and that these tests can complement more specific tests. Future trials are necessary to assess the value of these tests in multiparametric analysis, as well as their pathophysiological relevance.

Immunoproteome of Aspergillus fumigatus Using Sera of Patients with Invasive Aspergillosis

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Emylli D. Virginio

2014-08-01

Full Text Available Invasive aspergillosis is a life-threatening lung or systemic infection caused by the opportunistic mold Aspergillus fumigatus. The disease affects mainly immunocompromised hosts, and patients with hematological malignances or who have been submitted to stem cell transplantation are at high risk. Despite the current use of Platelia™ Aspergillus as a diagnostic test, the early diagnosis of invasive aspergillosis remains a major challenge in improving the prognosis of the disease. In this study, we used an immunoproteomic approach to identify proteins that could be putative candidates for the early diagnosis of invasive aspergillosis. Antigenic proteins expressed in the first steps of A. fumigatus germination occurring in a human host were revealed using 2-D Western immunoblots with the serum of patients who had previously been classified as probable and proven for invasive aspergillosis. Forty antigenic proteins were identified using mass spectrometry (MS/MS. A BLAST analysis revealed that two of these proteins showed low homology with proteins of either the human host or etiological agents of other invasive fungal infections. To our knowledge, this is the first report describing specific antigenic proteins of A. fumigatus germlings that are recognized by sera of patients with confirmed invasive aspergillosis who were from two separate hospital units.

Effects Of Amitryptilin Administration on Rat Sera and Brain Beta-endorphins

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Radivoj Jadrić

2006-11-01

Full Text Available The aim of our study was to establish the influence of antidepressive drugs on serum and brain beta-endorphins in experimental animals. Experiment was performed on albino Wistar rats. Antidepressant amitryptiline was used, and for quantification of sera and brain beta-endorphins RIA technique. Our results showed difference between sera and brain beta-endorphins concentration in amitryptiline pretreated animals, vs. those in serum and brain of control group treated with 0.95% NaCl. This study shows that use of psychoactive drugs have influence on sera and brain beta-endorphins concentration. Beta-endorphins could be of great importance, used as markers for evaluation of antidepressant drug effects.

Correlation between chemical components of billary calculi and bile & sera and bile of gallstone patients.

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Chandran, Prasheeda; Garg, Pradeep; Pundir, Chandra S

2005-07-01

Total cholesterol, total bilirubin, calcium, oxalate, inorganic phosphate, magnesium, iron, copper, sodium and potassium were analyzed quantitatively in gallstones, bile of gall bladder and sera of 200 patients of cholelithiasis (52 cholesterol, 76 mixed and 72 pigment stone patients) and their contents were correlated between calculi and bile and sera and bile in these three type of stone patients. A significant positive correlation was observed between total cholesterol, total bilirubin of calculi and bile, copper of bile and sera of cholesterol stone patients, copper of calculi and bile, total bilirubin, oxalate, magnesium, potassium of sera and bile of pigment stone patients and oxalate and iron of stone and bile, total bilirubin, oxalate, sodium of sera and bile of mixed stone patients. A significant negative correlation was found between magnesium of serum and bile of cholesterol stone patients, oxalate of calculi and bile of pigment stone patients and magnesium of serum and bile of mixed stone patients.

Assessing the clinical utility of measuring Insulin-like Growth Factor Binding Proteins in tissues and sera of melanoma patients

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Buckley Michael T

2008-11-01

Full Text Available Abstract Background Different Insulin-like Growth Factor Binding Proteins (IGFBPs have been investigated as potential biomarkers in several types of tumors. In this study, we examined both IGFBP-3 and -4 levels in tissues and sera of melanoma patients representing different stages of melanoma progression. Methods The study cohort consisted of 132 melanoma patients (primary, n = 72; metastatic, n = 60; 64 Male, 68 Female; Median Age = 56 prospectively enrolled in the New York University School of Medicine Interdisciplinary Melanoma Cooperative Group (NYU IMCG between August 2002 and December 2006. We assessed tumor-expression and circulating sera levels of IGFBP-3 and -4 using immunohistochemistry and ELISA assays. Correlations with clinicopathologic parameters were examined using Wilcoxon rank-sum tests and Spearman-rank correlation coefficients. Results Median IGFBP-4 tumor expression was significantly greater in primary versus metastatic patients (70% versus 10%, p = 0.01 A trend for greater median IGFBP-3 sera concentration was observed in metastatic versus primary patients (4.9 μg/ml vs. 3.4 μg/ml, respectively, p = 0.09. However, sera levels fell within a normal range for IGFBP-3. Neither IGFBP-3 nor -4 correlated with survival in this subset of patients. Conclusion Decreased IGFBP-4 tumor expression might be a step in the progression from primary to metastatic melanoma. Our data lend support to a recently-described novel tumor suppressor role of secreting IGFBPs in melanoma. However, data do not support the clinical utility of measuring levels of IGFBP-3 and -4 in sera of melanoma patients.

Characterization of sequence diversity in Plasmodium falciparum SERA5 from Indian isolates

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Rahul C.N

2015-06-01

Full Text Available Objective: To characterize the sequence diversity of blood-stage Plasmodium falciparum serine repeat antigen-5 (PfSERA5 which is lacking in a malaria-endemic country like India. Methods: In this study, parasitic DNA was obtained from field isolates collected from various geographic regions. Subsequently, PfSERA5 gene sequence was PCR amplified and DNA sequenced. Results: We reported the existence of unique repeat polymorphisms and novel haplotypes for both the octamer repeat (OR and serine repeat (SR regions of the N-terminal fragment of PfSERA5 from Indian isolates. Several isolates from India were identical to low-frequency African haplotypes. Unique finding of our study was an Indian isolate showing deletion in a perfectly conserved 14 mer sequence within octamer repeat. Indian haplotypes reported in this study were found to be distributed into the three earlier classified allelic clusters of FCR3, K1 and Honduras showcasing broad diversity as compared to worldwide haplotypes. Conclusions: This study is the first report on genetic diversity of PfSERA5 antigen from India. Further evaluation of these haplotypes by serotyping would provide useful information for investigating variant-specific immunity and aid in malaria vaccine research.

Confirmation of high specificity of an automated enzyme immunoassay test for serological diagnosis of syphilis: retrospective evaluation versus results after implementation.

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van Dommelen, Laura; Hoebe, Christian J P A; van Tiel, Frank H; Thijs, Carel; Goossens, Valère J; Bruggeman, Cathrien A; van Loo, Inge H M

2015-03-01

The optimal algorithm for serological syphilis screening is still a matter of debate. We have previously evaluated the performance of the Bioelisa Syphilis 3.0, using a selection of archived sera, and in this study compare these results with the Bioelisa results after clinical implementation. All Bioelisa Syphilis 3.0 results obtained since clinical implementation were analyzed. Bioelisa-positive or borderline samples were retested using Treponema pallidum particle agglutination, rapid plasma reagin test, fluorescent treponemal antibody-absorption test, and/or immunoblot. On sera sent in together with cerebrospinal fluid, occasionally both the T. pallidum particle agglutination and Bioelisa were performed. The Bioelisa was performed on 14,622 sera. Bioelisa-positive samples, which were not retested by the previously described assays, were withdrawn from the database (n = 36). In 1.3% of the samples (187/14,586), the Bioelisa was positive or borderline and, ultimately, 115 sera were considered true positive (prevalence 0.8%). The specificity of the Bioelisa was 99.5%. Based on the results of all performed diagnostic assays, the specificity of the Bioelisa of 99.5% is very consistent with that found in the initial study (100%; 95% confidence interval was 98.0%-100%). Interpreting (positive) test results is difficult in the absence of a gold standard, especially when the disease prevalence is low. Results should be viewed in the light of the patients' characteristics.

Immunodetection of Fasciola gigantica circulating antigen in sera of infected individuals for laboratory diagnosis of human fascioliasis.

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Attallah, Abdelfattah M; Bughdadi, Faisal A; El-Shazly, Atef M; Ismail, Hisham

2013-10-01

Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961, P 93%), and a significant correlation (r = 0.715, P fascioliasis.

Characterization of anti-liver-kidney microsome antibody (anti-LKM1) from hepatitis C virus-positive and -negative sera.

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Yamamoto, A M; Cresteil, D; Homberg, J C; Alvarez, F

1993-06-01

Hepatitis C virus-related antibodies were found in sera positive for antibodies to liver/kidney microsome antibody, usually considered a marker of autoimmune hepatitis. The aim of this study was to analyze the specificity of this autoantibody in sera from patients with and without hepatitis C virus infection. Fifteen anti-hepatitis C virus- and anti-liver kidney microsome-positive sera were compared with 11 sera from patients with autoimmune hepatitis, for reactivity against rat and human liver microsomal proteins, P450IID6 recombinant proteins, and various synthetic peptides spanning the 241-429 amino acids sequence of the P450IID6. Ten of 11 sera from patients with autoimmune hepatitis bound to recombinant proteins spanning the P450IID6 region between amino acids 72 and 458. These sera bound to the 254-271 peptide, and some also recognized the 321-351, 373-389 and 410-429 peptides. Four of 15 antihepatitis C virus recognized the fusion protein coded by the full-length P450IID6 complementary DNA; 3 of them also reacted with the P450IID6 region between amino acids 72-456. Only 1 sera recognized the 321-351 peptide. P450IID6 antigenic sites recognized by anti-hepatitis C virus-positive sera were different from those recognized by sera from patients with autoimmune hepatitis.

Demonstration of carbohydrate-specific immunoglobulin G4 antibodies in sera of patients receiving grass pollen immunotherapy

NARCIS (Netherlands)

van Ree, R.; Aalberse, R. C.

1995-01-01

From a group of 92 patients receiving grass pollen immunotherapy, and selected on grounds of high IgG4 titers against Lol p I, sera were tested for IgG4 antibodies against the glycosylated grass pollen allergen Lol p XI. In 72 of 92 cases IgG4 antibodies were demonstrated. The N-glycan of Lol p XI

Detection of IgE in the sera of rodents: comparison of the applicability of ELISA and RIA

Energy Technology Data Exchange (ETDEWEB)

Reiner, G; Zahner, H [Institut fuer Parasitologie der Justus-Liebig-Universitaet Giessen, Germany, F.R.; Haque, A [Institut Pasteur, 59 - Lille (France). Centre d' Immunologie et de Biologie Parasitaire

1984-04-13

RIA and ELISA were compared for their ability to detect IgE in different rodent species. With a sheep anti-rat IgE antibody good correlation (P < 0.001) between the 2 assay methods for IgE was found in rats. RIA failed to detect the IgE of Mastomys natalensis while ELISA proved to be a suitable test. However, both tests failed to measure IgE in sera of Nile rats.

SERA: Simulation Environment for Radiotherapy Applications - Users Manual Version 1CO

International Nuclear Information System (INIS)

Venhuizen, James Robert; Wessol, Daniel Edward; Wemple, Charles Alan; Wheeler, Floyd J; Harkin, G. J.; Frandsen, M. W.; Albright, C. L.; Cohen, M.T.; Rossmeier, M.; Cogliati, J.J.

2002-01-01

This document is the user manual for the Simulation Environment for Radiotherapy Applications (SERA) software program developed for boron-neutron capture therapy (BNCT) patient treatment planning by researchers at the Idaho National Engineering and Environmental Laboratory (INEEL) and students and faculty at Montana State University (MSU) Computer Science Department. This manual corresponds to the final release of the program, Version 1C0, developed to run under the RedHat Linux Operating System (version 7.2 or newer) or the Solaris Operating System (version 2.6 or newer). SERA is a suite of command line or interactively launched software modules, including graphical, geometric reconstruction, and execution interface modules for developing BNCT treatment plans. The program allows the user to develop geometric models of the patient as derived from Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) images, perform dose computation for these geometric models, and display the computed doses on overlays of the original images as three dimensional representations. This manual provides a guide to the practical use of SERA, but is not an exhaustive treatment of each feature of the code

SERA: Simulation Environment for Radiotherapy Applications - Users Manual Version 1CO

Energy Technology Data Exchange (ETDEWEB)

Venhuizen, James Robert; Wessol, Daniel Edward; Wemple, Charles Alan; Wheeler, Floyd J; Harkin, G. J.; Frandsen, M. W.; Albright, C. L.; Cohen, M.T.; Rossmeier, M.; Cogliati, J.J.

2002-06-01

This document is the user manual for the Simulation Environment for Radiotherapy Applications (SERA) software program developed for boron-neutron capture therapy (BNCT) patient treatment planning by researchers at the Idaho National Engineering and Environmental Laboratory (INEEL) and students and faculty at Montana State University (MSU) Computer Science Department. This manual corresponds to the final release of the program, Version 1C0, developed to run under the RedHat Linux Operating System (version 7.2 or newer) or the Solaris™ Operating System (version 2.6 or newer). SERA is a suite of command line or interactively launched software modules, including graphical, geometric reconstruction, and execution interface modules for developing BNCT treatment plans. The program allows the user to develop geometric models of the patient as derived from Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) images, perform dose computation for these geometric models, and display the computed doses on overlays of the original images as three dimensional representations. This manual provides a guide to the practical use of SERA, but is not an exhaustive treatment of each feature of the code.

Antibodies from the sera of HIV-infected patients efficiently hydrolyze all human histones.

Science.gov (United States)

Baranova, Svetlana V; Buneva, Valentina N; Nevinsky, Georgy A

2016-08-01

Histones and their post-translational modifications have key roles in chromatin remodeling and gene transcription. Besides intranuclear functions, histones act as damage-associated molecular pattern molecules when they are released into the extracellular space. Administration of exogenous histones to animals leads to systemic inflammatory and toxic responses through activating Toll-like receptors and inflammasome pathways. Here, using ELISA it was shown that sera of HIV-infected patients and healthy donors contain autoantibodies against histones. Autoantibodies with enzymic activities (abzymes) are a distinctive feature of autoimmune diseases. It was interesting whether antibodies from sera of HIV-infected patients can hydrolyze human histones. Electrophoretically and immunologically homogeneous IgGs were isolated from sera of HIV-infected patients by chromatography on several affinity sorbents. We present first evidence showing that 100% of IgGs purified from the sera of 32 HIV-infected patients efficiently hydrolyze from one to five human histones. Several rigid criteria have been applied to show that the histone-hydrolyzing activity is an intrinsic property of IgGs of HIV-infected patients. The relative efficiency of hydrolysis of histones (H1, H2a, H2b, H3, and H4) significantly varied for IgGs of different patients. IgGs from the sera of 40% of healthy donors also hydrolyze histones but with an average efficiency approximately 16-fold lower than that of HIV-infected patients. Similar to proteolytic abzymes from the sera of patients with several autoimmune diseases, histone-hydrolyzing IgGs from HIV-infected patients were inhibited by specific inhibitors of serine and of metal-dependent proteases, but an unexpected significant inhibition of the activity by specific inhibitor of thiol-like proteases was also observed. Because IgGs can efficiently hydrolyze histones, a negative role of abzymes in development of acquired immune deficiency syndrome cannot be

Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

Science.gov (United States)

Attallah, Abdelfattah M.; Bughdadi, Faisal A.; El-Shazly, Atef M.

2013-01-01

Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961, P 93%), and a significant correlation (r = 0.715, P fascioliasis. PMID:23945158

21 CFR 864.2800 - Animal and human sera.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Animal and human sera. 864.2800 Section 864.2800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture Products § 864.2800 Animal and...

Influence of previous experience on resistance training on reliability of one-repetition maximum test.

Science.gov (United States)

Ritti-Dias, Raphael Mendes; Avelar, Ademar; Salvador, Emanuel Péricles; Cyrino, Edilson Serpeloni

2011-05-01

The 1-repetition maximum test (1RM) has been widely used to assess maximal strength. However, to improve accuracy in assessing maximal strength, several sessions of the 1RM test are recommended. The aim of this study was to analyze the influence of previous resistance training experience on the reliability of 1RM test. Thirty men were assigned to the following 2 groups according to their previous resistance training experience: no previous resistance training experience (NOEXP) and more than 24 months of resistance training experience (EXP). All subjects performed the 1RM tests in bench press and squat in 4 sessions on distinct days. There was a significant session × group effect in bench press (F = 3.09; p reliability of the 1RM test is influenced by the subject's previous experience in resistance training. Subjects without experience in resistance training require more practice and familiarization and show greater increases in maximal strength between sessions than subjects with previous experience in resistance training.

[Evaluation of performance and false positivity of Mediace RPR test that uses a chemistry autoanalyzer].

Science.gov (United States)

Noh, Jaekwang; Ko, Hak Hyun; Yun, Yeomin; Choi, Young Sook; Lee, Sang Gon; Shin, Sue; Han, Kyou Sup; Song, Eun Young

2008-08-01

We evaluated the performance and false positive rate of Mediace RPR test (Sekisui, Japan), a newly introduced nontreponemal test using a chemistry autoanalyzer. The sensitivity of Mediace RPR test was analyzed using sera from 50 patients with syphilis in different stages (8 primary, 7 secondary, and 35 latent), 14 sera positive with fluorescent treponemal antibody absorption (FTA-ABS) IgM, and 74 sera positive with conventional rapid plasma regain (RPR) card test (Asan, Korea) and also positive with Treponema pallidum hemagglutination (TPHA) test or FTA-ABS IgG test. The specificity was analyzed on 108 healthy blood donors. We also performed RPR card test on 302 sera that had been tested positive with Mediace RPR test and also performed TPHA or FTA-ABS IgG test to analyze the false positive rate of Mediace RPR test. A cutoff value of 0.5 R.U. (RPR unit) was used for Mediace RPR test. Mediace RPR test on syphilitic sera of different stages (primary, secondary, and latent stages) and FTA-ABS IgM positive sera showed a sensitivity of 100%, 100%, 82.9% and 100%, respectively. Among the 74 sera positive with conventional RPR card test and TPHA or FTA-ABS IgG test, 55 were positive with Mediace test. The specificity of Mediace RPR test on blood donors was 97.2%. Among the 302 sera positive with Mediace RPR test, 137 sera (45.4%) were negative by RPR card and TPHA/FTA-ABS IgG tests. Although the sensitivities of Mediace RPR were good for primary and secondary syphilis, due to its high negative rate of Mediace RPR over the conventional RPR positive samples, further studies are necessary whether it can replace conventional nontreponemal test for screening purpose. Moreover, in view of the high false positive rate, positive results by Mediace RPR test should be confirmed with treponemal tests.

Detection of Potentially Diagnostic Leishmania Antigens with Western Blot Analysis of Sera from Patients with Cutaneous and Visceral Leishmaniases

Directory of Open Access Journals (Sweden)

Seyyed Javad SEYYEDTABAEI

2017-06-01

Full Text Available Background: Visceral leishmaniasis (VL and cutaneous leishmaniasis (CL are important public health problems in Iran. We aimed to evaluate the diagnostic potential of Western blot (WB compared with indirect immunofluorescence test (IFAT to serodiagnosis of leishmaniasis.Methods: This study was performed from 2010-2014 and participants were different parts of Iran. Serum samples were obtained from 43 patients with proven CL, 33 patients with proven VL, 39 patients with other parasitic diseases and 23 healthy individuals. Results: WB sensitivity for CL and VL was 100% and 91%, compared to IFA 4.6% and 87.8%, respectively. Sera from patients with CL and VL recognized numerous antigens with molecular weights ranging from 14 to 68 kDa and 12 to 94 kDa, respectively. The most sensitive antigens were 14 and 16 kDa for CL recognized by 100% of the sera from patients with proven CL and 12, 14 and 16 kDa for VL, recognized by 63.6%, 100% and 63.6% of the sera from patients with proven VL respectively. WB analysis is more sensitive than IFAT for the diagnosis of leishmaniasis particularly in cases of cutaneous leishmaniasis. The 12, 14 and 16 kDa can be valuable diagnostic molecules for serodiagnosis of leishmaniasis because at least two immunogenic molecules were simultaneously detected by all patient sera, as well as produced antibodies against these antigens have no cross-reactivity with other control groups.Conclusion: WB could be useful for screening and serodiagnosis of CL and VL in epidemiologic studies in endemic areas.

Cassava leaves in combination with sera onggok and rice bran as supplements in buffaloes ration

International Nuclear Information System (INIS)

Hendratno, C.; Sofian, L.A.; Abidin, Z.; Bahaudin, R.; Suharyono.

1988-01-01

Two experiments have been undertaken to evaluate the utilization of cassava leaves in combination with sera onggok or rice bran as supplements in buffalo ration under traditional village condition. In experiment 1, 16 buffaloes were divided in four groups, each receiving a different ration ranging from mixed forage alone to mixed forage supplemented with a combination of cassava leaves and sera onggok or rice bran. Changes in dry metter consumption, daily weight gain, feed convertion ratio and incom over feed cost were assesed. Experiments 2 covered an in vitro study on the changes in rumen fermentation as affected by different rations. The results of experiment 1 indicated the lack of differences in dry matter consumption. However, the daily weight gain, feed convertion ratio and income over feed cost (IOFC) higher in animal receiving mixed forage suplement with cassava leaves in combination with either sera onggok or rice bran as compared to those of animal receiving mixed forage or mixed forage supplemented with cassava leaves. Experiment 2 revealed that amonia concentration and volatile fatty acid production were able to support a higher microbil activity supplemented with cassava leaves in combination with either sera onggok or rice bran as compared to those receiving the other two rations. In conclusion it is obvious that cassava leaves in combination with either sera onggok or rice bran used as supplements could promote a better production in animal in the villages. (author). 7 refs, 1 fig, 5 tabs

Identification of streptococcal proteins reacting with sera from Behçet's disease and rheumatic disorders.

Science.gov (United States)

Cho, Sung Bin; Lee, Ju Hee; Ahn, Keun Jae; Cho, Suhyun; Park, Yong-Beom; Lee, Soo-Kon; Bang, Dongsik; Lee, Kwang Hoon

2010-01-01

We evaluated the reactivity of sera from Behçet's disease (BD), systemic lupus erythematosus (SLE), dermatomyositis (DM), rheumatoid arthritis (RA), and Takayasu's arteritis (TA) patients against human α-enolase and streptococcal α-enolase, and identified additional streptococcal antigens. Enzyme-linked immunosorbent assay (ELISA) and immunoblotting were performed using sera from patients with BD, SLE, DM, RA, and TA and healthy volunteers (control) against human α-enolase and streptococcal α-enolase. Immunoblot analysis and matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry were used to identify and recombine other streptococcal antigens. Specific positive signals against recombinant human α-enolase were detected by IgM ELISA of serum samples from 50% of BD, 14.3% of SLE, 57.1% of DM, 42.9% of RA, and 57.1% of TA patients. Specific positive signals against streptococcal α-enolase were detected from 42.9% of BD, 14.3% of DM, and 14.3% of TA patients. No SLE and RA sera reacted against streptococcal α-enolase antigen. Streptococcal proteins reacting with sera were identified as hypothetical protein (HP) for SLE and DM patients, acid phosphatase (AP) for RA patients, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for TA patients. We observed that RA patients did not present serum reactivity against either HP or GAPDH though BD, SLE, DM, and TA patients did. Also, AP reacted with sera from BD, SLE, DM, RA, and TA patients.

Sample handling for mass spectrometric proteomic investigations of human sera.

NARCIS (Netherlands)

West-Nielsen, M.; Hogdall, E.V.; Marchiori, E.; Hogdall, C.K.; Schou, C.; Heegaard, N.H.H.

2005-01-01

Proteomic investigations of sera are potentially of value for diagnosis, prognosis, choice of therapy, and disease activity assessment by virtue of discovering new biomarkers and biomarker patterns. Much debate focuses on the biological relevance and the need for identification of such biomarkers

Biofilm and planktonic pneumococci demonstrate disparate immunoreactivity to human convalescent sera

Directory of Open Access Journals (Sweden)

Shivshankar Pooja

2011-11-01

Full Text Available Abstract Background Streptococcus pneumoniae (the pneumococcus is the leading cause of otitis media, community-acquired pneumonia (CAP, sepsis, and meningitis. It is now evident that S. pneumoniae forms biofilms during nasopharyngeal colonization; the former which facilitates persistence, the latter, a prerequisite for subsequent development of invasive disease. Proteomic evaluation of S. pneumoniae suggests the antigen profile available for host-recognition is altered as a consequence of biofilm growth. This has potentially meaningful implications in regards to adaptive immunity and protection from disseminated disease. We therefore examined the antigen profile of biofilm and planktonic pneumococcal cell lysates, tested their reactivity with human convalescent sera and that generated against biofilm pneumococci, and examined whether immunization with biofilm pneumococci protected mice against infectious challenge. Results Biofilm pneumococci have dramatically altered protein profiles versus their planktonic counterparts. During invasive disease the humoral immune response is skewed towards the planktonic protein profile. Immunization with biofilm bacteria does not elicit a strong-cross-reactive humoral response against planktonic bacteria nor confer resistance against challenge with a virulent isolate from another serotype. We identified numerous proteins, including Pneumococcal serine-rich repeat protein (PsrP, which may serve as a protective antigens against both colonization and invasive disease. Conclusion Differential protein production by planktonic and biofilm pneumococci provides a potential explanation for why individuals remain susceptible to invasive disease despite previous colonization events. These findings also strongly suggest that differential protein production during colonization and disease be considered during the selection of antigens for any future protein vaccine.

Detection of toxoplasma gondii antigens in sera from experimentally infected mice

International Nuclear Information System (INIS)

Shojaee, S.; Keshavarz, H.; Rezaian, M.; Mohebali, M.

2007-01-01

Detection of Toxoplasma antigen in serum of mice by Immunoblotting. strain. IgG isolated from rabbits that were immunized with T. gondii Immunoblotting was performed to detect T. gondii antigens in sera of mice. Serum samples from mice experimentally infected with T. gondii RH strain. The value of Immunoblotting in diagnosis of toxoplasmosis in acute stage of infection. The antigen bands detected in serum sample of mice were experimentally infected with T. gondii tachyzoite in immunoblotting. Six bands demonstrated on seventh post infection day six bands were identified. Similarly on sixth day four bands, on day five three bands and on fourth post infection day two bands were identified. No band was detected in control group sera. Immunoblotting is a sensitive method for diagnosis of acute stage of toxoplasmosis. (author)

Anti-leptospirose agglutinins in equine sera, from São Paulo, Goias, and Mato Grosso do Sul, Brazil, 1996-2001

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H. Langoni

2004-01-01

Full Text Available Equine leptospirosis can present a non-symptomatic form, an acute clinical form, or even develop chronically, causing reproductive alterations, such as abortion and recurrent uveitis. Since the prevalence of leptospirosis in several countries and regions is widely reported, the objective of this study was to verify the prevailing equine leptospirosis in different regions of Brazil. Sera from 1402 blood samples from horses of different age, sex, breed, and purpose were examined. These samples came from southeastern and central west states of Brazil. The method utilized was the Microscopic Agglutination Test (MAT, with 12 different Leptospira serovars. From the sera tested, 754 (54% were positive for one (385 or more (372 serovars. These results were higher when compared to national and international levels. The most commonly found serovars were icterohaemorrhagiae (37.01%, suggesting exposure to rodents, castellonis (16.97%, and djasiman (15.19%. There were significant differences of reagents between sexes, and a tendency toward higher positivity with age. Distribution of sera-reagents related to aptitude showed a markedly higher value for work animals than for sporting ones. Higher rates were found for animals with undefined breed. There were no significant differences related to regional origin. As an indication of multiple exposure, significant associations were observed between the following serovars: castellonis and djasiman; castellonis and grippotyphosa; castellonis and copenhageni; castellonis and icterohaemorrhagiae; castellonis and pomona; canicola and pomona; canicola and djasiman; djasiman and copenhageni; icterohaemorrhagiae and djasiman; icterohaemorrhagiae and pyrogenes; copenhageni and pomona. These results showed the necessity of further studies on the epidemiology of this disease in equines and its relationship to human illness.

Correlation between chemical components of billary calculi and bile & sera and bile of gallstone patients

OpenAIRE

Chandran, Prasheeda; Garg, Pradeep; Pundir, Chandra S.

2005-01-01

Total cholesterol, total bilirubin, calcium, oxalate, inorganic phosphate, magnesium, iron, copper, sodium and potassium were analyzed quantitatively in gallstones, bile of gall bladder and sera of 200 patients of cholelithiasis (52 cholesterol, 76 mixed and 72 pigment stone patients) and their contents were correlated between calculi and bile and sera and bile in these three type of stone patients. A significant positive correlation was observed between total cholesterol, total bilirubin of ...

9th International Conference on Software Engineering Research, Management and Applications(SERA 2011)

CERN Document Server

Software Engineering Research, Management and Applications 2011

2012-01-01

The purpose of the 9th International Conference on Software Engineering Research, Management and Applications(SERA 2011) held on August 10-12, 2011 in Baltimore, Maryland was to bring together scientists, engineers, computer users, and students to share their experiences and exchange new ideas and research results about all aspects (theory, applications and tools) of computer and information sciences, and to discuss the practical challenges encountered along the way and the solutions adopted to solve them.   The conference organizers selected 12 outstanding papers from SERA 2011, all of which you will find in this volume of Springer’s Studies in Computational Intelligence.

14C-Metampicillin stability in several physiological sera

International Nuclear Information System (INIS)

Jimeno de Osso, F.; Casas Medica, F.; Carriazo Tovar, D.

1981-01-01

Degradation of 14 C -metampicillin incorporated to several physiological sera for medical uses has been studied. Influence of environmental conditions as well as possible interaction with the solvent have been specially analyzed. Degradation level of the labelled multiplication has been determined and its degradation products have been separated by using chromatographic and radiochemical methods. Likewise, the 14 C -metam picill synthesis has been described. Finally, the results obtained have been discussed and evaluated. (Author) 9 refs

Allergy to insect stings. IV. Diagnosis by radioallergosorbent test (R.A.S.T.)

International Nuclear Information System (INIS)

Sobotka, A.K.; Adkinson, N.F. Jr.; Valentine, M.D.; Lichtenstein, L.M.

1978-01-01

Radioallergosorbent tests (RAST(s)) have been developed and assessed for the diagnosis of insect hypersensitivity by using a purified allergen from honeybee venom, phospholipase A, and crude yellow jacket venom. Sera from 193 patients positive both by history and skin test to one of these insects were compared with various groups of control sera. Eighty percent of sera from skin test-positive patients were RAST positive; positive RAST were found in 16% of sera tested from skin test-negative patients. A highly positive RAST correlates well with a positive skin test and clinical sensitivity, but serum IgE is not measurable in many patients with mast cell or basophil bound antibody. Since biologically important reactions of antigen with IgE require that the antibody be cell bound, skin testing would be preferred to RAST if one were limited to a single test for the diagnosis of insect allergy

A population screening test for antibody to measles virus

International Nuclear Information System (INIS)

Friedman, M.G.

1981-01-01

In areas where sporadic cases of measles continue to occur in spite of vaccination programs, the availability of a simple screening test for determination of seropositivity to measles virus is desirable. A sensitive radioimmunoassay (RIA) screening test (ST) for the detection of IgG antibody to measles virus, based on a solid phase RIA, is described. The assays were performed on polyvinyl microtiter plates for which the RIAST requires only 5 μl of serum per subject. Antigen consisted of a sonicated extract of measles virus-infected Vero cells. Rabbit antihuman IgG specific for the Fc-segment of human IgG, labelled with 125 I, was used to detect human IgG bound to viral antigen. The basic RIA method was characterized by carrying out full titrations of sera of 53 healthy adults, 10 children, and 13 patients with measles-associated illness. These sera were also tested by the hemagglutination inhibition (HI) technique; most of the measles sera were also tested by complement fixation (CF). RIAST results (expressed as binding ratios) obtained for 52 healthy adults are compared with their RIA serum titers. Of the 200 sera of patients of various ages tested by the RIAST, 63 borderline sera were also tested by HI. The RIAST, which does not require serum treatment other than inactivation, proved to be more sensitive as an indicator of seropositivity than HI. Implications of the results and practical applications of the screening test are discussed. (author)

SERA Scenarios of Early Market Fuel Cell Electric Vehicle Introductions: Modeling Framework, Regional Markets, and Station Clustering

Energy Technology Data Exchange (ETDEWEB)

Bush, B. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Melaina, M. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Penev, M. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Daniel, W. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

2013-09-01

This report describes the development and analysis of detailed temporal and spatial scenarios for early market hydrogen fueling infrastructure clustering and fuel cell electric vehicle rollout using the Scenario Evaluation, Regionalization and Analysis (SERA) model. The report provides an overview of the SERA scenario development framework and discusses the approach used to develop the nationwidescenario.

Influence of previous knowledge in Torrance tests of creative thinking

OpenAIRE

Aranguren, María; Consejo Nacional de Investigaciones Científicas y Técnicas CONICET

2015-01-01

The aim of this work is to analyze the influence of study field, expertise and recreational activities participation in Torrance Tests of Creative Thinking (TTCT, 1974) performance. Several hypotheses were postulated to explore the possible effects of previous knowledge in TTCT verbal and TTCT figural university students’ outcomes. Participants in this study included 418 students from five study fields: Psychology;Philosophy and Literature, Music; Engineering; and Journalism and Advertisin...

The Plasmodium serine-type SERA proteases display distinct expression patterns and non-essential in vivo roles during life cycle progression of the malaria parasite.

Science.gov (United States)

Putrianti, Elyzana D; Schmidt-Christensen, Anja; Arnold, Iris; Heussler, Volker T; Matuschewski, Kai; Silvie, Olivier

2010-06-01

Parasite proteases play key roles in several fundamental steps of the Plasmodium life cycle, including haemoglobin degradation, host cell invasion and parasite egress. Plasmodium exit from infected host cells appears to be mediated by a class of papain-like cysteine proteases called 'serine repeat antigens' (SERAs). A SERA subfamily, represented by Plasmodium falciparum SERA5, contains an atypical active site serine residue instead of a catalytic cysteine. Members of this SERAser subfamily are abundantly expressed in asexual blood stages, rendering them attractive drug and vaccine targets. In this study, we show by antibody localization and in vivo fluorescent tagging with the red fluorescent protein mCherry that the two P. berghei serine-type family members, PbSERA1 and PbSERA2, display differential expression towards the final stages of merozoite formation. Via targeted gene replacement, we generated single and double gene knockouts of the P. berghei SERAser genes. These loss-of-function lines progressed normally through the parasite life cycle, suggesting a specialized, non-vital role for serine-type SERAs in vivo. Parasites lacking PbSERAser showed increased expression of the cysteine-type PbSERA3. Compensatory mechanisms between distinct SERA subfamilies may thus explain the absence of phenotypical defect in SERAser disruptants, and challenge the suitability to develop potent antimalarial drugs based on specific inhibitors of Plasmodium serine-type SERAs.

Near-infrared Raman spectroscopy to detect anti-Toxoplasma gondii antibodies in blood sera of domestic cats

Science.gov (United States)

Duarte, Janaina; Pacheco, Marcos T. T.; Silveira, Landulfo, Jr.; Machado, Rosangela Z.; Martins, Rodrigo A. L.; Zangaro, Renato A.; Villaverde, Antonio G. J. B.

2001-05-01

Near-infrared (NIR) Raman spectroscopy has been studied for the last years for many biomedical applications. It is a powerful tool for biological materials analysis. Toxoplasmosis is an important zoonosis in public health, cats being the principal responsible for the transmission of the disease in Brazil. The objective of this work is to investigate a new method of diagnosis of this disease. NIR Raman spectroscopy was used to detect anti Toxoplasma gondii antibodies in blood sera from domestic cats, without sample preparation. In all, six blood serum samples were used for this study. A previous serological test was done by the Indirect Immunoenzymatic Assay (ELISA) to permit a comparative study between both techniques and it showed that three serum samples were positive and the other three were negative to toxoplasmosis. Raman spectra were taken for all the samples and analyzed by using the principal components analysis (PCA). A diagnosis parameter was defined from the analysis of the second and third principal components of the Raman spectra. It was found that this parameter can detect the infection level of the animal. The results have indicated that NIR Raman spectroscopy, associated to the PCA can be a promising technique for serological analysis, such as toxoplasmosis, allowing a fast and sensitive method of diagnosis.

Radioimmunoassay of creatine kinase-B isoenzyme in human sera: results in patients with acute myocardial infarction

International Nuclear Information System (INIS)

Willerson, J.T.; Stone, M.J.; Ting, R.; Mukherjee, A.; Gomez-Sanchez, C.E.; Lewis, P.; Hersh, L.B.

1977-01-01

A radiommunoassay was developed to measure serum levels of the B isoenzyme of creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2) (CPK) in order to evaluate the time course and frequency of MB isoenzyme elevation in patients with acute myocardial infarction. The method can identify as little as 0.2 ng of the B portion of the CPK-MB isoenzyme, does not significantly crossreact with CPK-MM isoenzyme, and is not affected by storage of serum at -20 0 . CPK isoenzyme containing B subunits was detected in 48 out of 51 sera from normal adults; serum levels in these individuals ranged between 1.2 and 12.5 ng/ml[mean +- SEM was 2.7 +- 0.30 ng/ml]. The mean serum level of CPK-B isoenzyme in a pool of sera obtained from 100 normal subjects was 2.9 +- 0.35 ng/ml; two patients with rhabdomyolysis that were studied had serum CPK-B isoenzyme levels of 2.5 and 3.5 ng/ml, respectively. In contrast, serum levels of the CPK-B isoenzyme were markedly elevated in sera from 18 patients with acute myocardial infarcts when obtained within 12 hr after hospital admission; the mean +- SEM concentration was 56 +- 7.8 ng/ml. We performed serial determinations on 14 patients with acute myocardial infarcts and demonstrated that maximal serum CPK-B levels occurred within the first 12 hr after admission and were lower thereafter. The serum concentration of B-containing CPK isoenzyme in 19 additional patients admitted with chest pain but without acute myocardial infarction was 3.4 +- 0.50 ng/ml. Thus, radioimmunoassay measurement of CPK-B isoenzyme appears to be a useful and sensitive test for the detection of acute myocardial infarcts in patients

SERA -- An advanced treatment planning system for neutron therapy and BNCT

International Nuclear Information System (INIS)

Nigg, D.W.; Wemple, C.A.; Wessol, D.E.; Wheeler, F.J.; Albright, C.; Cohen, M.; Frandsen, M.; Harkin, G.; Rossmeier, M.

1999-01-01

Detailed treatment planning calculations on a patient-specific basis are required for boron neutron capture therapy (BNCT). Two integrated treatment planning systems developed specifically for BNCT have been in clinical use in the United States over the past few years. The MacNCTPLAN BNCT treatment planning system is used in the clinical BNCT trials that are underway at the Massachusetts Institute of Technology. A second system, BNCT rtpe (BNCT radiation therapy planning environment), developed independently by the Idaho national Engineering and Environmental Laboratory (INEEL) in collaboration with Montana State University (MSU), is used for treatment planning in the current series of BNCT clinical trials for glioblastoma at Brookhaven National Laboratory (BNL). This latter system is also licensed for use at several other BNCT research facilities worldwide. Although the currently available BNCT planning systems have served their purpose well, they suffer from somewhat long computation times (2 to 3 CPU-hours or more per field) relative to standard photon therapy planning software. This is largely due to the need for explicit three-dimensional solutions to the relevant transport equations. The simplifying approximations that work well for photon transport computations are not generally applicable to neutron transport computations. Greater computational speeds for BNCT treatment planning must therefore generally be achieved through the application of improved numerical techniques rather than by simplification of the governing equations. Recent efforts at INEEL and MSU have been directed toward this goal. This has resulted in a new paradigm for this type of calculation and the subsequent creation of the new simulation environment for radiotherapy applications (SERA) treatment planning system for BNCT. SERA is currently in initial clinical testing in connection with the trials at BNL, and it is expected to replace the present BNCT rtpe system upon general release

Coronative antibody tires in sera of healthy adults and experimentally infected volunteers.

Science.gov (United States)

Bradburne, A F; Somerset, B A

1972-06-01

Six coronaviruses isolated in the U.S.A. have been inoculated into volunteers and all produced colds. Between 10 and 20% of infected volunteers developed heterologous antibody responses after these and other experimental infections with coronaviruses. The haemagglutination-inhibition test with the OC43 virus strain was found to detect antibody rises after infection with a variety of strains.Studies on normal adult sera taken between 1965 and 1970 revealed a high frequency of neutralizing antibody to one strain (229 E) and a frequency of HI antibody to strain OC43 which fluctuated from year to year. Complement-fixing antibodies to these two viruses were also found, revealing an apparent increase in the activity of coronaviruses in the general population of the U.K., during the winter of 1968-9.

Cross-reactive anti-PfCLAG9 antibodies in the sera of asymptomatic parasite carriers of Plasmodium vivax

Science.gov (United States)

Costa, Joana D'Arc Neves; Zanchi, Fernando Berton; Rodrigues, Francisco Lurdevanhe da Silva; Honda, Eduardo Rezende; Katsuragawa, Tony Hiroschi; Pereira, Dhélio Batista; Taborda, Roger Lafontaine Mesquita; Tada, Mauro Shugiro; Ferreira, Ricardo de Godoi Mattos; Pereira-da-Silva, Luiz Hildebrando

2013-01-01

The PfCLAG9 has been extensively studied because their immunogenicity. Thereby, the gene product is important for therapeutics interventions and a potential vaccine candidate. Antibodies against synthetic peptides corresponding to selected sequences of the Plasmodium falciparum antigen PfCLAG9 were found in sera of falciparum malaria patients from Rondônia, in the Brazilian Amazon. Much higher antibody titres were found in semi-immune and immune asymptomatic parasite carriers than in subjects suffering clinical infections, corroborating original findings in Papua Guinea. However, sera of Plasmodium vivax patients from the same Amazon area, in particular from asymptomatic vivax parasite carriers, reacted strongly with the same peptides. Bioinformatic analyses revealed regions of similarity between P. falciparum Pfclag9 and the P. vivax ortholog Pvclag7. Indirect fluorescent microscopy analysis showed that antibodies against PfCLAG9 peptides elicited in BALB/c mice react with human red blood cells (RBCs) infected with both P. falciparum and P. vivax parasites. The patterns of reactivity on the surface of the parasitised RBCs are very similar. The present observations support previous findings that PfCLAG9 may be a target of protective immune responses and raises the possibility that the cross reactive antibodies to PvCLAG7 in mixed infections play a role in regulate the fate of Plasmodium mixed infections. PMID:23440122

Autoantibodies in breast cancer sera are not epiphenomena and may participate in carcinogenesis

International Nuclear Information System (INIS)

Fernández Madrid, Félix; Maroun, Marie-Claire; Olivero, Ofelia A; Long, Michael; Stark, Azadeh; Grossman, Lawrence I; Binder, Walter; Dong, Jingsheng; Burke, Matthew; Nathanson, S David; Zarbo, Richard; Chitale, Dhananjay; Zeballos-Chávez, Rocío; Peebles, Carol

2015-01-01

The objective of this work was to demonstrate that autoantibodies in breast cancer sera are not epiphenomena, and exhibit unique immunologic features resembling the rheumatic autoimmune diseases. We performed a comprehensive study of autoantibodies on a collection of sera from women with breast cancer or benign breast disease, undergoing annual screening mammography. All women in this study had suspicious mammography assessment and underwent a breast biopsy. We used indirect immunofluorescence, the crithidia assay for anti-dsDNA antibodies, and multiple ELISAs for extractable nuclear antigens. Autoantibodies were detected in virtually all patients with breast cancer, predominantly of the IgG1 and IgG3 isotypes. The profile detected in breast cancer sera showed distinctive features, such as antibodies targeting mitochondria, centrosomes, centromeres, nucleoli, cytoskeleton, and multiple nuclear dots. The majority of sera showing anti-mitochondrial antibodies did not react with the M2 component of pyruvate dehydrogenase, characteristic of primary biliary cirrhosis. Anti-centromere antibodies were mainly anti-CENP-B. ELISAs for extractable nuclear antigens and the assays for dsDNA were negative. The distinctive autoantibody profile detected in BC sera is the expression of tumor immunogenicity. Although some of these features resemble those in the rheumatic autoimmune diseases and primary biliary cirrhosis, the data suggest the involvement of an entirely different set of epithelial antigens in breast cancer. High titer autoantibodies targeting centrosomes, centromeres, and mitochondria were detected in a small group of healthy women with suspicious mammography assessment and no cancer by biopsy; this suggests that the process triggering autoantibody formation starts in the pre-malignant phase and that future studies using validated autoantibody panels may allow detection of breast cancer risk in asymptomatic women. Autoantibodies developing in breast cancer are not

Effects of sera obtained from electrically charged human body on action potential of giant axon of squid and its relationship to the therapy of the atomic bomb sequela, (2)

International Nuclear Information System (INIS)

Hirofuji, Michio; Hatashita, Toshiyuki; Takemura, Hideyuki; Oda, Nobuo.

1984-01-01

The giant axon of squid was perfused for 20 min with sea water and four kinds of mixture of sera and sea water (1:2), and spike potential of the axon was compared by using a computer. Perfusates used were sea water, sera obtained before electric charge to the human body (pre-sera), sera obtained from the human body electrically charged with -300 volt (negative sera), and sera obtained from the human body electrically charged with +300 volt (positive sera). Negative sera increased action potential of the axon, and positive sera decreased action potential of the axon. These results revealed that negative sera have a greater deal of e - , and positive sera have less quantity of e - than pre-sera, suggesting the involvement of e - in the action potential of the axon. Microtubules in the inner part of the axonal membrane and cell membrane seem to be most greatly related to e - ; however, changes in the other axons, cell membrane and protoplasm should also be taken into account. These experimental results seem to be of great value, particularly providing useful information on the treatment for late effects (cell damage) of atomic bombing or burn. (Namekawa, K.)

Direct assessment of cumulative aryl hydrocarbon receptor agonist activity in sera from experimentally exposed mice and environmentally exposed humans

DEFF Research Database (Denmark)

Schlezinger, Jennifer J; Bernard, Pamela L; Haas, Amelia

2010-01-01

(PCB)-exposed Faroe Islanders using an AhR-driven reporter cell line. To validate relationships between serum AhR agonist levels and biological outcomes, AhR agonist activity in mouse sera correlated with toxic end points. AhR agonist activity in unmanipulated ("neat") human sera was compared......, was associated with the frequency of recent pilot whale dinners, but did not correlate with levels of PCBs quantified by GC/MS. Surprisingly, significant "baseline" AhR activity was found in commercial human sera. CONCLUSIONS: An AhR reporter assay revealed cumulative levels of AhR activation potential in neat...

Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species.

Science.gov (United States)

Yu, Meng; Stevens, Vicky; Berry, Jody D; Crameri, Gary; McEachern, Jennifer; Tu, Changchun; Shi, Zhengli; Liang, Guodong; Weingartl, Hana; Cardosa, Jane; Eaton, Bryan T; Wang, Lin-Fa

2008-02-29

Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and diagnostic tests. Although there have been many reports of such work done for SARS-CoV, these were mainly focused on the immune responses of humans and mice. In this study, we aim to search for and compare immunodominant regions of the spike (S) and nucleocapsid (N) proteins which are recognized by sera from different animal species, including mouse, rat, rabbit, civet, pig and horse. Twelve overlapping recombinant protein fragments were produced in Escherichia coli, six each for the S and N proteins, which covered the entire coding region of the two proteins. Using a membrane-strip based Western blot approach, the reactivity of each antigen fragment against a panel of animal sera was determined. Immunodominant regions containing linear epitopes, which reacted with sera from all the species tested, were identified for both proteins. The S3 fragment (aa 402-622) and the N4 fragment (aa 220-336) were the most immunodominant among the six S and N fragments, respectively. Antibodies raised against the S3 fragment were able to block the binding of a panel of S-specific monoclonal antibodies (mAb) to SARS-CoV in ELISA, further demonstrating the immunodominance of this region. Based on these findings, one-step competition ELISAs were established which were able to detect SARS-CoV antibodies from human and at least seven different animal species. Considering that a large number of animal species are known to be susceptible to SARS-CoV, these assays will be a useful tool to trace the origin and transmission of SARS-CoV and to minimise the risk of animal-to-human transmission.

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

Circadian rhythms of cysteine proteinases and cystatins, potential tumour markers, in normal sera

International Nuclear Information System (INIS)

Cimerman, N.; Krasovec, M.; Mesko-Brguljan, P.; Suskovic, S.; Kos, J.

2002-01-01

Circadian day/night variations have been evidenced in all major groups of organisms and at all levels of organisation of the organism. Circadian intra-individual variations are known for a number of analyses in serum including tumour-associated markers. It was suggested that the serum levels of cysteine proteinases and their inhibitors may be of clinical importance for prognosis and diagnosis in cancer. Since known circadian rhythms are important for choosing the best sampling time, interpretation of the results of a diagnostic test, patient monitoring, and timing of a therapy, our objective was to establish 24-h variations of cysteine proteinases, cathepsins B, H, L, and their low molecular weight inhibitors, stefin A, stefin B, and cystatin C, in sera from healthy subjects. (author)

LIQUID CHROMATOGRAPHIC / MASS SPECTROMETRIC DETERMINATION OF MORPHINE, CODEINE AND COCAINE IN HUMAN SERA USING A NEW INTERNAL SURFACE REVERSED PHASE COLUMN

OpenAIRE

Hattori, Hideki; Arinobu, Tetsuya; Iwai, Masae; Suzuki, Osamu; Seno, Hiroshi

2003-01-01

A new internal surface reversed phase column (Shim-pack MAYI-ODS) was used for analysis of morphine, codeine and cocaine in human sera by LC/MS/MS. The column enabled direct injection of crude biological samples without pretreatment realizing a rapid analytical procedure. Therecoveries of morphine, codeine and cocaine spiked into human sera were 50-60 %. The regression equations for morphine, codeine and cocaine in sera, using atropine as internal standard, showed good linearity in the ranges...

Exosomes isolated from cancer patients' sera transfer malignant traits and confer the same phenotype of primary tumors to oncosuppressor-mutated cells.

Science.gov (United States)

Abdouh, Mohamed; Hamam, Dana; Gao, Zu-Hua; Arena, Vincenzo; Arena, Manuel; Arena, Goffredo Orazio

2017-08-30

Horizontal transfer of malignant traits from the primary tumor to distant organs, through blood circulating factors, has recently become a thoroughly studied metastatic pathway to explain cancer dissemination. Recently, we reported that oncosuppressor gene-mutated human cells undergo malignant transformation when exposed to cancer patients' sera. We also observed that oncosuppressor mutated cells would show an increased uptake of cancer-derived exosomes and we suggested that oncosuppressor genes might protect the integrity of the cell genome by blocking integration of cancer-derived exosomes. In the present study, we tested the hypothesis that cancer patients' sera-derived exosomes might be responsible for the malignant transformation of target cells and that oncosuppressor mutation would promote their increased uptake. We also sought to unveil the mechanisms behind the hypothesized phenomena. We used human BRCA1 knockout (BRCA1-KO) fibroblasts as target cells. Cells were treated in vitro with cancer patients' sera or cancer patients' sera-derived exosomes. Treated cells were injected into NOD-SCID mice. Immunohistochemical analyses were performed to determine the differentiation state of the xenotransplants. Mass spectrometry analyses of proteins from cancer exosomes and the BRCA1-KO fibroblasts' membrane were performed to investigate possible de novo expression of molecules involved in vesicles uptake. Blocking of the identified molecules in vitro was performed and in vivo experiments were conducted to confirm the role of these molecules in the malignant transformation carried out by cancer-derived exosomes. Cells treated with exosomes isolated from cancer patients' sera underwent malignant transformation and formed tumors when transplanted into immunodeficient mice. Histological analyses showed that the tumors were carcinomas that differentiated into the same lineage of the primary tumors of blood donors. Oncosuppressor mutation promoted the de novo expression

Reactions of chicken sera to recombinant Campylobacter jejuni flagellar proteins.

Science.gov (United States)

Yeh, Hung-Yueh; Hiett, Kelli L; Line, John E

2015-03-01

Campylobacter jejuni is a Gram-negative spiral rod bacterium and is the leading but underreported bacterial food-borne pathogen that causes human campylobacteriosis worldwide. Raw or undercooked poultry products are regarded as a major source for human infection. C. jejuni flagella have been implicated in colonization and adhesion to the mucosal surface of chicken gastrointestinal tracts. Therefore, flagellar proteins would be the excellent targets for further investigation. In this report, we used the recombinant technology to generate a battery of C. jejuni flagellar proteins, which were purified by His tag affinity chromatography and determined antigenic profiles of these recombinant flagellar proteins using sera from chickens older than 6 weeks of age. The immunoblot results demonstrate that each chicken serum reacted to various numbers of recombinant flagellar proteins. Among these recombinant proteins, chicken sera reacted predominantly to the FlgE1, FlgK, FlhF, FliG and FliY proteins. These antibody screening results provide a rationale for further evaluation of these recombinant flagellar proteins as potential vaccines for chickens to improve food safety as well as investigation of host immune response to C. jejuni.

The need for a sufficient number of low level sera in comparisons of different serum vitamin B12 assays

International Nuclear Information System (INIS)

Gijzen, A.H.J.; Kock, H.W. de; Meulendijk, P.N.; Schmidt, N.A.; Schopman, W.; Tertoolen, J.F.W.; Voogd, C.E.

1983-01-01

Eight radiochemical methods for the assay of vitamin B 12 in serum were compared with the microbiological assay with Lactobacillus leichmannii ATCC 7830 using 198 individual sera of patients. There was a good agreement between the results of most samples with some kits and the microbiological assay. However, especially in the sera of vitamin B 12 -deficient patients large discrepancies between the results could occur. These variations were due to both the kits used and the performance of the assays in different laboratories. A sufficient number of non-pooled sera of vitamin B 12 -deficient patients should be included in investigations to validate radiochemical methods. (Auth.)

Trace elements in sera of patients with hepatitis B: Determination and analysis

Science.gov (United States)

Saod, Wahran M.; Darwish, Nadiya T.; Zaidan, Tahseen A.; Alfalujie, Abdul Wahab A.

2018-04-01

Chronic Hepatitis B (HBV) is the leading cause of morbidity and mortality worldwide with about 248 million people having HBV infection. Trace elements e.g. copper (Cu), zinc (Zn), selenium (Se) and iron (Fe) are constituent components of many metal proteins and metalloenzymes in human sera. Therefore, the ratios of these trace elements in human sera are often stated to be a good marker for diagnosing various diseases including HBV. The aims of this study are: to compare the level of trace elements in sera of patients infected with HBV and healthy participants, and to evaluate the efficiency of analytical techniques (e.g. Inductively Coupled Plasma-Mass spectrometry (ICP-MS), Atomic Absorption Spectroscopy (hydride generation) (AAS) and Graphite Furnace Atomic Absorption Spectroscopy (GFAAS) that are currently used to detect Fe and Se elements in Patients' human sera. The findings of this study show that the concentration range of copper element between (132.80±28.64 µg/dl) to (105.66±23.20 µg/dl) was significantly higher in HBV infected patients as compared to those in healthy controls (91.27±9.20 µg/dl). Iron concentration range between (206.64±61.60 µg/l) to (170.00±36.71 µg/l) was significantly higher in HBV infected patients as compared to those in healthy controls (158.00±15.13 µg/l). However, patients with HBV had significantly lower serum concentrations of zinc with a concentration range between (111.64±20.90 µg/dl) to (99.25±24.06 µg/dl) as compared to those in healthy controls (113.44±16.38 µg/dl). While selenium concentration range between (64.39±7.39 µg/l) to (51.10±4.96 µg/l) was significantly lower in HBV infected patients as compared to those in healthy controls (67.68±7.60) (μg/l). Moreover, the results of this study suggest that (AAS) technique was the most accurate method to measure the concentration of selenium element, while (UV and ICP-MS) analytical techniques have the same efficiency in measuring the iron concentration.

Influence of Previous Knowledge in Torrance Tests of Creative Thinking

Directory of Open Access Journals (Sweden)

María Aranguren

2015-07-01

Full Text Available The aim of this work is to analyze the influence of study field, expertise and recreational activities participation in Torrance Tests of Creative Thinking (TTCT, 1974 performance. Several hypotheses were postulated to explore the possible effects of previous knowledge in TTCT verbal and TTCT figural university students’ outcomes. Participants in this study included 418 students from five study fields: Psychology;Philosophy and Literature, Music; Engineering; and Journalism and Advertising (Communication Sciences. Results found in this research seem to indicate that there in none influence of the study field, expertise and recreational activities participation in neither of the TTCT tests. Instead, the findings seem to suggest some kind of interaction between certain skills needed to succeed in specific studies fields and performance on creativity tests, such as the TTCT. These results imply that TTCT is a useful and valid instrument to measure creativity and that some cognitive process involved in innovative thinking can be promoted using different intervention programs in schools and universities regardless the students study field.

Sera of patients with recurrent miscarriages containing anti-trophoblast antibodies (ATAB) reduce hCG and progesterone production in trophoblast cells in vitro.

Science.gov (United States)

von Schönfeldt, Viktoria; Rogenhofer, Nina; Ruf, Katharina; Thaler, Christian J; Jeschke, Udo

2016-09-01

Reproductive failure including RM has been suggested to correlate with antibodies that cross react with HLA-negative syncytiotrophoblasts and we have reported that 17% of women with 2 or more miscarriages and 34% of women with 3 or more miscarriages express anti-trophoblast antibodies (ATAB). Until now, the mechanism, how ATAB interfere with pregnancy success is not known. HCG and progesterone both play fundamental roles in supporting human pregnancy. Therefore we investigated the effects of sera of RM patients containing ATAB on the hCG and progesterone production of cells of the choriocarcinoma cell line JEG-3. In vitro study to investigate effects of patient sera with and without ATAB on hCG and progesterone secretion of JEG-3 cells. The presence of ATAB was detected as described earlier. Effects of sera from ATAB positive and ATAB negative RM patients on hCG and progesterone secretion by JEG-3 cells were analysed 12 and 24h after plating. Sera of women without pregnancy pathologies served as controls. Sera of ATAB-positive RM patients significantly inhibit hCG secretion of JEG-3 cells for 12h after plating compared to sera of healthy controls (p=0.019) and significantly reduce progesterone production for 12h (p=0.046) and 24h (p=0.027) of co-culture. Sera of ATAB-negative RM patient show no significant effect on progesterone secretion. Inhibition of hCG and progesterone production might point to a mechanism, how ATAB interfere with early pregnancies. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Fingolimod prevents blood-brain barrier disruption induced by the sera from patients with multiple sclerosis.

Directory of Open Access Journals (Sweden)

Hideaki Nishihara

Full Text Available OBJECTIVE: Effect of fingolimod in multiple sclerosis (MS is thought to involve the prevention of lymphocyte egress from lymphoid tissues, thereby reducing autoaggressive lymphocyte infiltration into the central nervous system across blood-brain barrier (BBB. However, brain microvascular endothelial cells (BMECs represent a possible additional target for fingolimod in MS patients by directly repairing the function of BBB, as S1P receptors are also expressed by BMECs. In this study, we evaluated the effects of fingolimod on BMECs and clarified whether fingolimod-phosphate restores the BBB function after exposure to MS sera. METHODS: Changes in tight junction proteins, adhesion molecules and transendothelial electrical resistance (TEER in BMECs were evaluated following incubation in conditioned medium with or without fingolimod/fingolimod-phosphate. In addition, the effects of sera derived from MS patients, including those in the relapse phase of relapse-remitting (RR MS, stable phase of RRMS and secondary progressive MS (SPMS, on the function of BBB in the presence of fingolimod-phosphate were assessed. RESULTS: Incubation with fingolimod-phosphate increased the claudin-5 protein levels and TEER values in BMECs, although it did not change the amount of occludin, ICAM-1 or MelCAM proteins. Pretreatment with fingolimod-phosphate restored the changes in the claudin-5 and VCAM-1 protein/mRNA levels and TEER values in BMECs after exposure to MS sera. CONCLUSIONS: Pretreatment with fingolimod-phosphate prevents BBB disruption caused by both RRMS and SPMS sera via the upregulation of claudin-5 and downregulation of VCAM-1 in BMECs, suggesting that fingolimod-phosphate is capable of directly modifying the BBB. BMECs represent a possible therapeutic target for fingolimod in MS patients.

Detection of Toxoplasma gondii DNA in sera samples of mice experimentally infected

Directory of Open Access Journals (Sweden)

H. Langoni

2006-04-01

Full Text Available Detection of Toxoplasma gondii (T. gondii DNA in blood can help to diagnose the disease in its acute phase; however, it must be considered that hemoglobin, present in blood, can inhibit polymerase activity, making impracticable the detection of DNA in samples. Mice were experimentally infected via oral route with ME49 and BTU2 strains cysts and RH strain tachyzoites; polymerase chain reaction was used to detect T. gondii DNA in mice sera 18, 24, 48, 96, and 192 hours post infection (PI. Toxoplama gondii DNA was detected in only one animal infected with BTU2 strain, genotype III (isolated from a dog with neurological signs 18 hours PI. The agent's DNA was not detected in any sample of the other experimental groups. New studies must be carried out to verify the technique sensitivity in researches on this agent's genetic material using sera samples of acute-phase toxoplasmosis patients, especially in cases of immunosuppression.

Standardization and application of the solid phase C1q radioimmunoassay using soluble tetanus toxoid-antitetanus immune complexes in sera of patients with chronic polyarthritis and Lupus erythematodes

International Nuclear Information System (INIS)

Menzel, E.J.; Steffen, C.; Smolen, J.

1982-01-01

Soluble tetanus-antitetanus immune complexes were prepared with affinity-chromatography and gel chromatography. Serial dilutions of these immune complex preparations were tested in a solid phase C1q radioimmunoassay. Soluble immune complexes as well as aggregated human gamma globulin of identical protein concentrations were comparatively investigated. Soluble immune complexes rendered a more sensitive standardization of RIA. According to these observations a relation between μg/ml equivalents of defined tetanus-antitetanus complexes and ng second antibody obtained in C1q-RIA was calculated. Upper limit of mean values and two standard deviations of ng second antibody obtained in investigations of 55 normal sera was designated as 1 unit immune complexes and regarded as border line of negative results. Multiplication of μg/ml immune complex equivalents of 1 unit led to a scale of 1 to 15 units, showing the area of positive results. According to these values a standardization curve was constructed allowing a conversion of ng-second antibody obtained in serum investigations into immune complex units equivalent to defined standard immune complexes. With this curve investigation results of 56 RA sera and 21 SLE sera were expressed in the range of units, making a distinct gradation of positive results and a clear cut separation of positive and negative results possible. SLE sera of patients in acute stage showed highly positive results. (orig.) [de

Standardization and application of the solid phase C1q radioimmunoassay using soluble tetanus toxoid-antitetanus immune complexes in sera of patients with chronic polyarthritis and Lupus erythematodes

Energy Technology Data Exchange (ETDEWEB)

Menzel, E J; Steffen, C; Smolen, J

1982-11-22

Soluble tetanus-antitetanus immune complexes were prepared with affinity-chromatography and gel chromatography. Serial dilutions of these immune complex preparations were tested in a solid phase C1q radioimmunoassay. Soluble immune complexes as well as aggregated human gamma globulin of identical protein concentrations were comparatively investigated. Soluble immune complexes rendered a more sensitive standardization of RIA. According to these observations a relation between ..mu..g/ml equivalents of defined tetanus-antitetanus complexes and ng second antibody obtained in C1q-RIA was calculated. Upper limit of mean values and two standard deviations of ng second antibody obtained in investigations of 55 normal sera was designated as 1 unit immune complexes and regarded as border line of negative results. Multiplication of ..mu..g/ml immune complex equivalents of 1 unit led to a scale of 1 to 15 units, showing the area of positive results. According to these values a standardization curve was constructed allowing a conversion of ng-second antibody obtained in serum investigations into immune complex units equivalent to defined standard immune complexes. With this curve investigation results of 56 RA sera and 21 SLE sera were expressed in the range of units, making a distinct gradation of positive results and a clear cut separation of positive and negative results possible. SLE sera of patients in acute stage showed highly positive results.

Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts.

Science.gov (United States)

Wareth, Gamal; Eravci, Murat; Weise, Christoph; Roesler, Uwe; Melzer, Falk; Sprague, Lisa D; Neubauer, Heinrich; Murugaiyan, Jayaseelan

2016-04-30

Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B.) species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with either laboratory-grown species or vaccine strains, in the present study, field strains were utilized for analysis. The label-free quantitative proteomic analysis of the naturally isolated strains of these two closely related species revealed 402 differentially expressed proteins, among which 63 and 103 proteins were found exclusively in the whole cell extracts of B. abortus and B. melitensis field strains, respectively. The sera from four different naturally infected host species, i.e., cattle, buffalo, sheep, and goat were applied to identify the immune-binding protein spots present in the whole protein extracts from the isolated B. abortus and B. melitensis field strains and resolved on two-dimensional gel electrophoresis. Comprehensive analysis revealed that 25 proteins of B. abortus and 20 proteins of B. melitensis were distinctly immunoreactive. Dihydrodipicolinate synthase, glyceraldehyde-3-phosphate dehydrogenase and lactate/malate dehydrogenase from B. abortus, amino acid ABC transporter substrate-binding protein from B. melitensis and fumarylacetoacetate hydrolase from both species were reactive with the sera of all the tested naturally infected host species. The identified proteins could be used for the design of serological assays capable of detecting pan-Brucella, B. abortus- and B. melitensis-specific antibodies.

Express immunochromatographic detection of antibodies against Brucella abortus in cattle sera based on quantitative photometric registration and modulated cut-off level.

Science.gov (United States)

Sotnikov, Dmitriy V; Byzova, Nadezhda A; Zherdev, Anatoly V; Eskendirova, Saule Z; Baltin, Kairat K; Mukanov, Kasim K; Ramankulov, Erlan M; Sadykhov, Elchin G; Dzantiev, Boris B

2015-01-01

An immunochromatographic test system was developed for rapid detection of the levels of specific IgG antibodies to Brucella abortus lipopolysaccharide, as a tool for diagnosis of brucellosis in cattle. The pilot test strips were examined using blood sera from sick (78 samples) and healthy (35 samples) cows. The results obtained by immunochromatographic assay, using a portable optical densitometer for digital video detection, correlate well with the results obtained by immunoenzyme assay and are in agreement with the results of the disease diagnosis. The new test system allows detection of antibodies within 10 min and can be proposed as an alternative to the methods available for serodiagnosis of brucellosis.

Clinical Comparison of the Treponema pallidum CAPTIA Syphilis-G Enzyme Immunoassay with the Fluorescent Treponemal Antibody Absorption Immunoglobulin G Assay for Syphilis Testing

OpenAIRE

Halling, V. W.; Jones, M. F.; Bestrom, J. E.; Wold, A. D.; Rosenblatt, J. E.; Smith, T. F.; Cockerill, F. R.

1999-01-01

Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphilis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results of rapid plasmid reagin (RPR) testing of the same specimens. Borderline CAPTI...

Replacement of the in vivo neutralisation test for efficacy demonstration of tetanus vaccines ad us. vet.

Science.gov (United States)

Rosskopf, Ute; Noeske, Kerstin; Werner, Esther

2005-01-01

The bacterium Clostridium (C.) tetani is an ubiquitous pathogen. This anaerobic, gram-positive bacterium can form spores and can be found in the whole environment. It enters the body via injuries of the skin and wounds where it releases the neurotoxin "tetanospasmin" (= tetanus toxin). The animals most susceptible to tetanus infection are horses and sheep. Only active immunisation by tetanus vaccine provides effective protection against tetanus intoxication. The marketing authorisation requirements stipulate that efficacy of tetanus vaccines ad us. vet. must be demonstrated in all target animal species via determination of neutralising tetanus serum antitoxin concentrations. The standard method used for this purpose is still the toxin neutralisation test (TNT), as it quantifies the tetanus toxin-neutralising effect of tetanus serum antibodies in vivo. In this test, tetanus toxin is added to dilutions of serum from vaccinated horse and sheep. The serum dilutions are then administered to mice or guinea pigs, which are observed for toxic symptoms. Against the background of animal protection, the goal of one project of the Paul-Ehrlich-Institut (Bundesministerium fuer Bildung und Forschung (Federal Ministry for Education and Research), 0312636) was to establish an alternative to the toxin neutralisation test, enabling the testing of efficacy of tetanus vaccines with serological in vitro methods. For this purpose, a so-called double antigen ELISA (DAE) was established which enables the testing of sera of different species in one assay. In addition, the sera were tested in an indirect ELISA for horses and sheep separately. Altogether, ten groups of horses and eight groups of sheep were immunised with ten animals per group each. The tetanus vaccines comprised almost all products authorised for the German market at the start of the project. 564 horse sera and 257 sheep sera were tested using the two ELISA methods. Some sera were also tested in vivo. The kinetics of

An assessment of radioimmunoassay procedures for determination of anti-acetylcholine receptor antibodies in the sera of patients with myasthenia gravis

International Nuclear Information System (INIS)

Carter, B.; Harrison, R.; Lunt, G.G.; Morris, H.; Savage-Marengo, T.; Stephenson, F.A.

1981-01-01

A reproducible radioimmunoassay procedure for the determination of anti-acetylcholine receptor antibodies in the sera of patients with myasthenia gravis is described and examined in detail. The assay combines features of a number of methods previously outlined and allows repeat determinations of antibody titre in a given myasthenic serum sample with coefficient of variation 6%. The mean +- standard deviation for normal human serum anti-acetylcholine receptor antibodies was found by this procedure to be 0.024 +- 0.033 nmol/l α-bungarotoxin binding sites whereas the range for myasthenic patients was 0-139.14 nmol/l with a mean value of 7.55 nmol/l α-bungarotoxin binding sites. (author)

SYNTHESIS OF 25-HYDROXYVITAMIN D3 CONJUGATE WITH KEYHOLE LIMPET HEMOCYANIN AND OBTAINING OF IMMUNE SERA

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А. O. Mazanova

2015-08-01

Full Text Available The study was aimed at obtaining polyclonal antibodies that recognize 25-Hydroxyvitamin D3, for their further specifications and applications in immunochemical test systems of 25-Hydroxyvitamin D3, determination in blood serum. We used the methods of chemical synthesis of immunoconjugates (modified carbodiimide method using N´-ethyl carbodiimide, thin layer chromatography, gel filtration and indirect immunoenzyme analysis ELISA. The work describes the stages of the synthesis of 25-Hydroxyvitamin D3, immunoconjugate with keyhole limpet hemocyanin (KLH, which was used for receiving immune sera. As a result of mouse and rabbit immunization antisera were obtained and antibody titers against 25-Hydroxyvitamin D3, were tested by immunoenzyme assay. It was demonstrated that the titer of specific antibodies was higher in rabbits compared with mice. The resulting polyclonal antibodies can be used for the development of immunochemical test systems for screening studies of 25-Hydroxyvitamin D3, content in human blood serum as a marker of vitamin D3 availability.

Detection of antibodies to bacterial cell wall peptidoglycan in human sera

International Nuclear Information System (INIS)

Heymer, B.; Schleifer, K.H.; Read, S.; Zabriskie, J.B.; Krause, R.M.

1976-01-01

A radioimmunoassay has been developed for the measurement of antibodies to peptidoglycan in human sera including patients with rheumatic feaver and juvenile rheumatoid arthritis. The assay is based on the percentage of binding of the hapten 125 I-L-Ala-γ-D-Glu-L-Lys-D-Ala-D-Ala, the major peptide determinant of peptidoglycan. Because of differences in the avidity of the antibodies in different sera, the amount of antibody was expressed as pentapeptide hapten-binding capacity (pentapeptide-HBC in ng/ml of serum). Fourteen out of 105 normal blood donors had a pentapeptide-HBC value greater than or equal to 75 ng/ml serum. Values in healthy children 5 to 18 years of age were less than or equal to 50 ng/ml. Sixty-eight percent of the individuals with rheumatic fever had values greater than or equal to 75 ng/ml, an indication that streptococcal infections can stimulate an immune response to peptidoglycan. Thirty-five percent of the patients with juvenile rheumatoid arthritis had values greater than or equal to 75 ng/ml. Such a finding points to a possible association between bacterial infections and juvenile rheumatoid arthritis

Validating a bovine brucellosis elisa test for application in Uruguay

International Nuclear Information System (INIS)

Silva, M.; Muller, G.; Errico, F.; Dilave, M.

1998-01-01

Sera from 600 cattle on Rio Negro Island, known to be free of brucellosis, and 400 sera from vaccinated cattle but known to be negative in the Rose-Bengal test were selected for validation of the FAO/IAEA test kit for detection of antibody to Brucella abortus. Two conjugates, one a polyclonal antiserum and the other a monoclonal antibody, were evaluated. When evaluated for reproducibility using the sera from uninfected cattle, the average coefficient of variation for duplicate samples was 7.1%±5.5. The serum control samples did not exceed OD limits as established for the kit, for any of the 15 plates evaluated. When evaluated by regression analysis, the control sera had an average correlation coefficient of 0.996, indicating a high degree of agreement between the observed OD values of controls on each plate vs the expected values for those controls. Specificity in the assay was >98% as calculated by the PP or regression methods. Comparison of the monoclonal and polyclonal conjugates using sera from vaccinated cattle indicated that many of the cattle must have been vaccinated as adults because of high antibody levels detected by both conjugates. Before this assay can be used on vaccinated animals, the kit will have to be evaluated using sera from animals of known age of vaccination. (author)

Usefulness of a single-assay chemiluminescence test (Tularaemia VIRCLIA IgG + IgM monotest) for the diagnosis of human tularemia. Comparison of five serological tests.

Science.gov (United States)

Cubero, África; Durántez, Carlos; Almaraz, Ana; Fernández-Lago, Luis; Gutiérrez, María P; Castro, María J; Bratos, Miguel A; Simarro, María; March, Gabriel A; Orduña, Antonio

2018-04-01

The aim of this work was to ascertain the usefulness of a new commercially-available single-assay chemiluminescence test (CHT) for the diagnosis of human tularemia (Tularaemia VIRCLIA IgG + IgM monotest, Vircell, Santa Fe, Granada, Spain). A total of 773 sera from 773 patients including 364 initial sera from patients with diagnosed tularemia, patients with suspected tularemia not confirmed (100), healthy people (152), patients with serology positive to Brucella (97), patients diagnosed with other infectious diseases (30), and patients diagnosed with autoimmune diseases (30) were included. All sera were tested by CHT, "in-house" microagglutination test (MAT), immunochromatographic test (ICT) (Virapid Tularaemia, Vircell, Santa Fe Granada, Spain), and "in-house" ELISA IgG, and ELISA IgM. Of the total initial sera, 334 (sensitivity 91.8%) were positive in the CHT, 332 (sensitivity 91.2%) in the MAT, 330 (sensitivity 90.7%) in the ICT, and 328 (sensitivity 90.1%) in the ELISA IgG and ELISA IgM tests. The specificity of the CHT was 96.7%; of the MAT, 100%; of the ICT, 98.7%; and of the ELISA IgG and ELISA IgM, 97.4%. In the group of patients with serology positive to Brucella, at least 12.4% of sera were positive in tularemia tests (12.4% in ELISA IgM, 13.4% in MAT, 14.4% in ICT, and 15.5% in CHT and ELISA IgG). In conclusion, CHT presents a sensitivity and specificity in early diagnosis of human tularemia, similar to MAT, ICT, and ELISA IgG and ELISA IgM. Its single assay design allows lower costs, especially in areas of low endemicity or inter-epidemic periods.

Antibodies to a novel leptospiral protein, LruC, in the eye fluids and sera of horses with Leptospira-associated uveitis.

Science.gov (United States)

Verma, Ashutosh; Matsunaga, James; Artiushin, Sergey; Pinne, Marija; Houwers, Dirk J; Haake, David A; Stevenson, Brian; Timoney, John F

2012-03-01

Screening of an expression library of Leptospira interrogans with eye fluids from uveitic horses resulted in identification of a novel protein, LruC. LruC is located in the inner leaflet of the leptospiral outer membrane, and an lruC gene was detected in all tested pathogenic L. interrogans strains. LruC-specific antibody levels were significantly higher in eye fluids and sera of uveitic horses than healthy horses. These findings suggest that LruC may play a role in equine leptospiral uveitis.

IgG autoantibodies against interleukin 1 alpha in sera of normal individuals

DEFF Research Database (Denmark)

Svenson, M; Poulsen, L K; Fomsgaard, A

1989-01-01

A pool of human sera from healthy blood donors was found to interfere competitively with the binding of 125I-labelled human recombinant interleukin 1 alpha (rIL-1 alpha) to the murine T-cell line EL4. The interference was reversible at the cellular level, and direct binding of the ligand to serum...

Humoral response of paracoccidioidomycosis sera in hamsters with different Venezuelan isolates

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Lilian M Spencer

2009-09-01

Full Text Available Humoral response of paracoccidioidomycosis sera in hamsters with different Venezuelan isolates. Paracoccidioidomycosis (PCM is a progressive systemic mycosis caused by the fungus Paraccocidioides brasiliensis (Pb, endemic to Venezuela and Latin America. In this study, eight different Venezuelan isolates obtained from patients with PCM, were inoculated intraperitoneally in Syrian hamsters (Cricetus auratus and studied by immune-serum. Each strain was collected by gently scraping the surface of the culture medium (Sabouraud Dextrose Agar and suspended in 3ml of 0.15 M phosphate-buffered saline. The antigen obtained was called Paraccocidioides brasiliensis Crude Antigen (CAP. Immunoblotting results showed that the immune-sera from hamsters recognized at least 3 bands: one over 200 kDa, and two of 80 and 15-20 kDa. This study suggests that IgG anti-CAP can reveal a significant variability in the eight Venezuelan isolates. Sera from 88 infected hamsters were evaluated by ELISA with eight different CAPs and Western blot with CAP 37383. ELISA results showed that, the antigen of the virulent isolate 37383 had the highest percentage (38% of positivity, while the non-virulent isolate 1458 had the lowest one (13.6%. Furthermore, scanning densitometry revealed that the isolate 37383 had less bands than the non-virulent isolates. These results suggest that the ELISA test with CAP 37383 can detect circulating antibodies, and that this virulent isolate may be useful for the diagnosis of PCM, and to monitor disease responses to treatments. Rev. Biol. Trop. 57 (3: 505-513. Epub 2009 September 30.La Paracoccidioidomicosis (PCM, es una micosis sistémica causada por el hongo Paraccocidioides brasiliensis (Pb, endémica en Venezuela y Latino América. En este estudio ocho diferentes aislados venezolanos, obtenidos de pacientes con PCM, fueron inoculados intraperitonealmente en hámsteres y fueron estudiados por ELISA e inmunoblotting. Los antígenos obtenidos de P

Corynebacterium renale as a cause of reactions to the complement fixation test for Johne's disease

NARCIS (Netherlands)

Gilmour, N.J.L.; Goudswaard, J.

Complement fixation (C.F.) tests and fluorescent antibody (F.A.) tests were carried out on sera from rabbits inoculated with Corynebacterium renale and Mycobacterium johnei, and on sera from cattle with C. renale pyelonephritis and with Johne's disease. Cross-reactions were a feature of the C.F.

Comprehensive Identification of Immunodominant Proteins of Brucella abortus and Brucella melitensis Using Antibodies in the Sera from Naturally Infected Hosts

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Gamal Wareth

2016-04-01

Full Text Available Brucellosis is a debilitating zoonotic disease that affects humans and animals. The diagnosis of brucellosis is challenging, as accurate species level identification is not possible with any of the currently available serology-based diagnostic methods. The present study aimed at identifying Brucella (B. species-specific proteins from the closely related species B. abortus and B. melitensis using sera collected from naturally infected host species. Unlike earlier reported investigations with either laboratory-grown species or vaccine strains, in the present study, field strains were utilized for analysis. The label-free quantitative proteomic analysis of the naturally isolated strains of these two closely related species revealed 402 differentially expressed proteins, among which 63 and 103 proteins were found exclusively in the whole cell extracts of B. abortus and B. melitensis field strains, respectively. The sera from four different naturally infected host species, i.e., cattle, buffalo, sheep, and goat were applied to identify the immune-binding protein spots present in the whole protein extracts from the isolated B. abortus and B. melitensis field strains and resolved on two-dimensional gel electrophoresis. Comprehensive analysis revealed that 25 proteins of B. abortus and 20 proteins of B. melitensis were distinctly immunoreactive. Dihydrodipicolinate synthase, glyceraldehyde-3-phosphate dehydrogenase and lactate/malate dehydrogenase from B. abortus, amino acid ABC transporter substrate-binding protein from B. melitensis and fumarylacetoacetate hydrolase from both species were reactive with the sera of all the tested naturally infected host species. The identified proteins could be used for the design of serological assays capable of detecting pan-Brucella, B. abortus- and B. melitensis-specific antibodies.

Microbial transglutaminase treatment in pasta-production does not affect the immunoreactivity of gliadin with celiac disease patients' sera.

Science.gov (United States)

Ruh, Tobias; Ohsam, Jürgen; Pasternack, Ralf; Yokoyama, Keiichi; Kumazawa, Yoshiyuki; Hils, Martin

2014-07-30

The effect of microbial transglutaminase (MTG)-treatment of pasta-dough on the immunoreactivity with celiac disease patient's sera has been investigated. Modification by MTG has been proven by determination of the MTG reaction product ε-(γ-glutamyl)lysine (3.63 μmol/g protein), which was not detectable in non-MTG-treated pasta. Antigenicity has been analyzed by immunoblotting and ELISA using gliadin-extracts from pasta and MTG-treated pasta. Immunoblotting showed that the antibody-population (antigliadin antibodies and antideamidated gliadin antibodies) of the sera is specific for every individual patient. Immunoblotting and ELISA showed that there is no difference in immunoreactivity of gliadin extracted from pasta and MTG-pasta. Recognition pattern and intensity in Western blot as well as antibody titer has also been identical even for sera with a high antideamidated gliadin antibody titer. These results indicate no difference between pasta-gliadin and MTG-pasta-gliadin and especially no increased deamidation in pasta-gliadin by MTG-treatment.

Congenital heart block maternal sera autoantibodies target an extracellular epitope on the α1G T-type calcium channel in human fetal hearts.

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Linn S Strandberg

Full Text Available Congenital heart block (CHB is a transplacentally acquired autoimmune disease associated with anti-Ro/SSA and anti-La/SSB maternal autoantibodies and is characterized primarily by atrioventricular (AV block of the fetal heart. This study aims to investigate whether the T-type calcium channel subunit α1G may be a fetal target of maternal sera autoantibodies in CHB.We demonstrate differential mRNA expression of the T-type calcium channel CACNA1G (α1G gene in the AV junction of human fetal hearts compared to the apex (18-22.6 weeks gestation. Using human fetal hearts (20-22 wks gestation, our immunoprecipitation (IP, Western blot analysis and immunofluorescence (IF staining results, taken together, demonstrate accessibility of the α1G epitope on the surfaces of cardiomyocytes as well as reactivity of maternal serum from CHB affected pregnancies to the α1G protein. By ELISA we demonstrated maternal sera reactivity to α1G was significantly higher in CHB maternal sera compared to controls, and reactivity was epitope mapped to a peptide designated as p305 (corresponding to aa305-319 of the extracellular loop linking transmembrane segments S5-S6 in α1G repeat I. Maternal sera from CHB affected pregnancies also reacted more weakly to the homologous region (7/15 amino acids conserved of the α1H channel. Electrophysiology experiments with single-cell patch-clamp also demonstrated effects of CHB maternal sera on T-type current in mouse sinoatrial node (SAN cells.Taken together, these results indicate that CHB maternal sera antibodies readily target an extracellular epitope of α1G T-type calcium channels in human fetal cardiomyocytes. CHB maternal sera also show reactivity for α1H suggesting that autoantibodies can target multiple fetal targets.

Purification of a Fe-SOD excreted by Leishmania braziliensis for specific antibodies detection in Mexican human sera: Cutting-edge the knowledge

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Silvia Stefania Longoni

2016-06-01

Full Text Available Clinical diagnosis of leishmaniasis is highly complex, presenting a wide range of clinical manifestations, sometimes non-specific, and thus the epidemiological study and diagnostic need specific molecular markers for each Leishmania species. Leishmania spp. posses different Fe-SOD isoforms, one of which is excreted into the external milieu and, presenting immunogenic characteristics, is a very reliable molecular marker. Superoxide dismutases (SODs are antioxidant metal-enzymes responsible for the dismutation of superoxide ion into hydrogen peroxide and molecular oxygen, and it is considered an important virulence factor. In this manuscript we have purified the iron(Fe-SOD excreted by Leishmania braziliensis using ion-exchange and molecular-sieve chromatography and we have studied it as an antigen in serodiagnostic analyses in ELISA and Western blot techniques, testing 213 human sera from Mexico. Indeed, L. braziliensis Fe-SODe has been purified 123.26 times with a specific activity of about 893.66 U/mg of protein. Applying the purified enzymes in serological tests we found 17.84% sera positive. We have demonstrated that the purified enzyme is more sensitive than the non-purified ones and we also demonstrated, for the first time, the presence of antibodies against L. braziliensis, not the main species in the country, in human population from Hidalgo and Nuevo Leon States.

Aspects pertinent to the usefulness of a solid phase radio-immuno-sorbent assay for the detection of spermatozoa antibodies in sera of infertility patients

International Nuclear Information System (INIS)

Hinrichs-Reiche, I.

1987-01-01

The solid phase Radio-Immuno-Sorbent Assay (RISA) is a highly sensitive and valid test to detect 125-iodinetagged antibodies to spermatozoa that allows qualitative and quantitative evaluations of sperm-incapacitating immunglobulin Ig G in sera from patients believed to be infertile for immunological reasons. The study failed to reveal any correlations between the results of RISA and those of micro-sperm-agglutination or micro-sperm-immobilisation tests. There was a major body of evidence pointing to possible links between female isoimmunity and male autoimmunity. (TRV) [de

Evaluation of an Immunocapture-Agglutination Test (Brucellacapt) for Serodiagnosis of Human Brucellosis

Science.gov (United States)

Orduña, Antonio; Almaraz, Ana; Prado, Ana; Gutierrez, M. Purificación; Garcia-Pascual, Agustina; Dueñas, Ana; Cuervo, Milagros; Abad, Ramon; Hernández, Beatriz; Lorenzo, Belen; Bratos, Miguel A.; Torres, Antonio Rodriguez

2000-01-01

We evaluated the validity and the usefulness of a new test for the diagnosis of human brucellosis based on an immunocapture-agglutination technique. A total of 315 sera from 82 patients with a diagnosis of brucellosis, 157 sera from patients in whom brucellosis was suspected but not confirmed, and 412 sera from people living in rural areas with endemic brucellosis were studied. The seroagglutination test (SAT), Coombs anti-Brucella test, and Brucellacapt test were evaluated. All the initial sera from the 82 patients proved to be positive in Brucellacapt and Coombs tests, while only 75 (91.4%) were positive in the SAT. If a ≥1/160 diagnostic threshold titer was defined for the Brucellacapt test, Coombs test, and SAT, the sensitivities were 95.1, 91.5, and 65.8%, respectively. Taking the same diagnostic threshold titer for the 157 sera from the unconfirmed but suspected patients, the specificities of the Brucellacapt, Coombs, and SAT were 81.5, 96.2, and 100%, respectively; for the 412 control sera, the specificities were 99.0, 99.8, and 100%. The diagnostic efficiency (area below the receiver operating characteristic curve) of Brucellacapt was 0.987852 (95% confidence interval [CI], 0.95109 to 0.99286), very similar to the diagnostic efficiency of the Coombs test (0.97611; 95% CI, 0.94781 to 0.99146) and higher than that of SAT (0.91013; 95% CI, 0.86649 to 0.94317). The results of the Brucellacapt test were compared with those of the Coombs test (correlation coefficient, 0.956; P = 0.000) and SAT (correlation coefficient, 0.866; P = 0.000). The study shows very good correlation between the Brucellacapt and Coombs tests, with a high concordance between titers obtained in the two tests. Nevertheless, lower correlation and concordance were found between the Brucellacapt and Coombs tests when the results for titers of ≥1/160 were compared (0.692; P = 0.000). In acute brucellosis, the Brucellacapt and Coombs tests render positive titers of ≥1/160. When the titers

Effect of haemolysis and repeated freeze-thawing cycles on wild boar serum antibody testing by ELISA

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Boadella Mariana

2011-11-01

Full Text Available Abstract Background Monitoring wildlife diseases is needed to identify changes in disease occurrence. Wildlife blood samples are valuable for this purpose but are often gathered haemolysed. To maximise information, sera often go through repeated analysis and freeze-thaw cycles. Herein, we used samples of clean and haemolysed Eurasian wild boar (Sus scrofa serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV. Results The estimated prevalence of antibodies against ADV was 50-53% for clean and haemolysed sera. Hence, haemolysis did not reduce the mean observed serum antibody prevalence. However, 10 samples changed their classification after repeated freeze-thawing. This included 3 (15% of the clean sera and 7 (41% of the haemolysed sera. Conclusions We recommend (1 establishing more restrictive cut-off values when testing wildlife sera, (2 recording serum quality prior to sample banking, (3 recording the number of freezing-thawing cycles and (4 store sera in various aliquots to reduce repeated usage. For instance, sera with more than 3 freeze-thaw cycles and a haemolysis of over 3 on a scale of 4 should better be discarded for serum antibody monitoring. Even clean (almost not haemolysed sera should not go through more than 5 freeze-thaw cycles.

Detection and measurement of antioxidant capacity in human sera

International Nuclear Information System (INIS)

Bognar, G.; Koeteles, G.J.; Otos, M.

1998-01-01

The total antioxidant capacity of human sera was measured by the Randox TAS assay and an average value of 1.55 mmol/L was found from 87 healthy adult persons. Exogenous antioxidant added to the blood could be measured additively. Upon X-irradiation of whole blood samples, the antioxidant value decreased down to 1 Gy linearly. Further decrease after higher doses, however, could not be detected. Reductions of radiation-induced human lymphocyte micronucleus frequency as a cytogenetic end-point were observed upon increasing the exogenous antioxidant level in serum with a water-soluble form of alpha-tocopherol, or a plant extract from Sylibum marianum L. in vitro. (author)

Serum neutralization as a differential serological test for classical swine fever virus and other pestivirus infections

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Paredes J.C.M.

1999-01-01

Full Text Available Serum neutralization tests (SN were performed against classical swine fever virus (CSFV, bovine viral diarrhea virus (BVDV and border disease virus (BDV on samples of swine serum collected for screening of antibodies to CSFV, in order to determine the SN value as a differential serological test. Ninety-nine sera out of a sample of 16,664 were positive for antibodies to pestiviruses in an ELISA test which did not distinguish antibodies to different pestiviruses. When submitted to SN, 81 sera were positive for CSFV antibodies only. In 17 sera, crossreactive antibodies to either CSFV, BVDV or BDV were detected. In most of these sera (13 out of 17 the differences between SN titres against the three viruses were not sufficient to estimate which was the most likely antibody-inducing virus. It was concluded that, for the SN to be useful in such differentiation, it is essential to examine a sample which must include a representative number of sera from the same farm where suspect animals were detected. When isolated serum samples are examined, such as those obtained with the sampling strategy adopted here, the SN may give rise to inconclusive results.

Study of FAO/IAEA/PANAFTOSA ELISA kit for the detection of antibodies against FMD

International Nuclear Information System (INIS)

Ravison, J.A.; Andrade Goncalves, D. de; Souza, R. de

1998-01-01

Two groups of sera were used to evaluate a liquid phase blocking ELISA (LPBE) for the detection of antibody against foot-and-mouth disease virus. One hundred and twenty sera, from animals with no previous history of FMD infection or vaccination, were analyzed by screening assay at a final dilution of 1:32. A second group of 120 sera, from animals vaccinated with an oil trivalent vaccine (O, A, C) were tested by titration in the LPBE. All the sera were tested against virus of three FMD serotypes, using O 1 Campos, A 24 Cruzeiro, C 3 Indaial virus strains. (author)

Total pepsin activity and gastrin in sera as markers of eradication of Helicobacter pylori

NARCIS (Netherlands)

Khoshkholgh, M.; Saberi-Firoozi, M.; Fattahi, M.; Siavoshi, F.; Khatibian, M.; Vahedi, H.; Mikaeli, J.; Ansari, R.; Alizadeh, B.; Malekzadeh, R.; Massarrat, S.

1994-01-01

The measurement of total pepsin activity by colorimetry, and gastrin by radioimmunoassay method was performed on the sera of 100 patients (80 with duodenal ulcer and 20 with non-ulcer dyspepsia) before and 4 weeks after the end of antibacterial treatment for eradication of Helicobacter pylori. While

A rapid, sensitive and reliable diagnostic test for scrub typhus in China

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Zhang Lijuan

2011-01-01

Full Text Available Purpose: To evaluate the performances for detection of IgM and IgG antibodies to Orientia. tsutsugamushi (Ot using a gold conjugate-based rapid diagnostic test (RDT. Materials and Methods: The RDT employing mixture recombinant 56-kDa proteins of O. tsutsugamushi and the mIFA assay was performed on 33 patients from Fujian and Yunnan province respectively and 94 positive sera (36 from Hainan province and 58 from Jiangsu province from convalescent stages of the patients with scrub typhus respectively and 82 negative sera from healthy farmers from Anhui province and Beijing City respectively in 2009. A comparison of the RDT and mIFA assay was performed by using the c2 test and the P level of ≤0.05 was considered to be significant. Results: Among these 94 positive sera from convalescent stages of the illness and 82 sera from control farmers, the specificity of RDT was 100% for both IgM and IgG tests. In 33 cases with scrub typhus, 5 cases were positively detected earlier by RDT than by mIFA for the IgM test, and 2 cases were positive for the IgG test. The sensitivities of RDT were 93.9% and 90.9% for IgM and IgG, respectively. Considering IgM and IgG together, the sensitivity was 100%. The geometric mean titre (GMT of IFA and the RDT assay in diluted sera from confirmed cases were 1:37 versus 1:113 respectively (P<0.001 for IgM test and 1:99 versus 1:279 respectively (P<0.016 for IgG. Conclusions: The RDT was more sensitive than the traditional IFA for the early diagnosis of scrub typhus and was particularly suitable for use in rural areas.

In Vitro Endothelial Cell Proliferation Assay Reveals Distinct Levels of Proangiogenic Cytokines Characterizing Sera of Healthy Subjects and of Patients with Heart Failure

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Rebecca Voltan

2014-01-01

Full Text Available Although myocardial angiogenesis is thought to play an important role in heart failure (HF, the involvement of circulating proinflammatory and proangiogenic cytokines in the pathogenesis and/or prognosis of HF has not been deeply investigated. By using a highly standardized proliferation assay with human endothelial cells, we first demonstrated that sera from older (mean age 52±7.6 years; n=46 healthy donors promoted endothelial cell proliferation to a significantly higher extent compared to sera obtained from younger healthy donors (mean age 29±8.6 years; n=20. The promotion of endothelial cell proliferation was accompanied by high serum levels of several proangiogenic cytokines. When we assessed endothelial cell proliferation in response to HF patients’ sera, we observed that a subset of sera (n=11 promoted cell proliferation to a significantly lesser extent compared to the majority of sera (n=18. Also, in this case, the difference between the patient groups in the ability to induce endothelial cell proliferation correlated to significant (P<0.05 differences in serum proangiogenic cytokine levels. Unexpectedly, HF patients associated to the highest endothelial proliferation index showed the worst prognosis as evaluated in terms of subsequent cardiovascular events in the follow-up, suggesting that high levels of circulating proangiogenic cytokines might be related to a worse prognosis.

Syphilis serology: Seroprevalence in a selected population and considerations on the Euroline WB test

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Andrea Amodeo

2010-06-01

Full Text Available Introduction: The clinical diagnosis of syphilis is always supported by appropriate laboratory tests and the test results are interpreted with reference to the patient’s history. In the diagnosis of syphilis, the use of tests based on antibody search that recognize both treponemal and reaginic antigens increases the diagnostic chances. Our study discusses the various serological and alternative tests currently available along with their limitations, and relates their results to the likely corresponding clinical stage of the disease. Methods: in our laboratory were analyzed 264 sera and 4 liquor (123 Females, 145 Males. 187 patients are subject at low risk for luetic infection, including pregnant woman, patient with organ transplant, outpatients or hospitalized undergoing routine serological, and 81 from patients with confirmed syphilis including 4 pregnant women in antibiotic treatment, patients with suspected disease, HIV positive and patients with autoimmune diseases with Cardiolipin positive. All sera were tested with ELISA Anti-Treponema pallidum Screen (IgG / IgM and in parallel with agglutination tests VDRL and TPHA. On all positive sera was tested Euroline-WB EUROIMMUN and reading done with the program EuroLineScan. Results: by ELISA Anti-Treponema pallidum Screen IgG / IgM 162 sera were negative and 106 sera positive (39.5%, distributed as follows: 45 (42% with a value greater than 200 RU / ml, 43 (41% with a value> 22 RU / ml and 18 (17% with a borderline value between> 16 to <22 RU / ml. The execution of the Blot IgG showed: 18 negative sera, 6 with borderline value with one only band of specific antigens (p15, p45, p47 or p17, while 82, including 4 liquor (neurolue, were certainly positive showing more than one band antibody to the treponemal antigens. Only one patient had in place at the time of screening, an initial infection; in fact, there was a single clear positivity in the IgM protein bands, while 7 sera was uncertain values

Antibody-dependent cellular cytotoxicity and neutralizing activity in sera of HIV-1-infected mothers and their children.

Science.gov (United States)

Broliden, K; Sievers, E; Tovo, P A; Moschese, V; Scarlatti, G; Broliden, P A; Fundaro, C; Rossi, P

1993-01-01

The prognostic and protective role of antibodies mediating cellular cytotoxicity (ADCC) and neutralization was evaluated in sera of HIV-1-infected mothers and their consecutively followed children. The presence and titres of ADCC mediating and/or neutralizing antibodies in maternal sera did not predict HIV-1 infection in their respective children. No significant difference in the sera from the children was seen when comparing the presence of neutralizing antibodies between the uninfected and infected children. Stratification of the infected group according to clinical status revealed differences. Only one of 24 AIDS patients had a high neutralizing titre against IIIB. Four patients had a very low titre and the remaining had no detectable neutralizing antibodies at all. In contrast, 10/17 infected non-AIDS children had neutralizing antibodies. Similarly, no significant difference was seen when comparing the presence of ADCC-mediating antibodies between the uninfected and the infected group of children. However, a significantly higher frequency of ADCC was seen in the seropositive non-AIDS children compared with the AIDS children. This study clearly shows that the presence of antibodies mediating ADCC and neutralization in infected children, 0-2 years old, is associated with a better clinical status and delayed disease progression. PMID:8324904

Functional MRI of the visual cortex and visual testing in patients with previous optic neuritis

DEFF Research Database (Denmark)

Langkilde, Annika Reynberg; Frederiksen, J.L.; Rostrup, Egill

2002-01-01

of the activated area and the signal change following ON, and compared the results with results of neuroophthalmological testing. We studied nine patients with previous acute ON and 10 healthy persons served as controls using fMRI with visual stimulation. In addition to a reduced activated volume, patients showed...... a reduced blood oxygenation level dependent (BOLD) signal increase and a greater asymmetry in the visual cortex, compared with controls. The volume of visual cortical activation was significantly correlated to the result of the contrast sensitivity test. The BOLD signal increase correlated significantly......The volume of cortical activation as detected by functional magnetic resonance imaging (fMRI) in the visual cortex has previously been shown to be reduced following optic neuritis (ON). In order to understand the cause of this change, we studied the cortical activation, both the size...

Evaluation of serological tests for the diagnosis of rickettsiosis in Denmark

DEFF Research Database (Denmark)

Kantsø, Bjørn; Svendsen, Claus Bo; Jørgensen, Charlotte Svaerke

2009-01-01

Two commercially available immunofluorescence assays (IFA) were compared using historical sera evaluated for rickettsial antibodies by the Weil-Felix test. An IFA test produced by Focus Diagnostics prepared with Rickettsia rickettsii and R. typhi antigens was compared with a custom made kit from....... When analyzing the data using the manufacturers' cut-off values, 41% of samples from presumably healthy blood donors were found positive for spotted fever group Rickettsia antibodies. This does not correlate to the general picture of rickettsiosis in Denmark. Furthermore, sera with Coxiella burnetii...

Parvovirus B19 infections in state of Rio de Janeiro, Brasil: 526 sera analyzed by IgM-enzyme-linked immunosorbent assay and polymerase chain reaction

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MCL Mendonça

2005-12-01

Full Text Available In this study were analyzed 526 sera; the patients aged from two days to 65 years old presenting exanthema, which was the most frequent symptom observed, besides fever, adenomegaly, and arthralgia. These sera were negative by enzyme-linked immunosorbent assay (IgM-ELISA for either rubella (495, toxoplasma (41, cytomegalovirus (12, measles (40, dengue (56, and they were submitted to nested polymerase chain reaction (PCR for B19 DNA and commercial IgM-ELISA for B19. In 39 abortion cases, IgM or DNA were not detected, therefore they were not took into account for analysis. Specific DNA and IgM were detected respectively in 71 (14.5% and IgM in 62 (12.7% sera from 487 sera analyzed. IgM and DNA were simultaneously detected in 43 (8.8%, while agreement among the results by PCR and IgM-ELISA was observed in 440 (90.4%. The sera were collected from January 1999 to December 2000, most of them in 1999 (325, during winter and spring. The major number of clinical cases was observed in the age group from one to ten years old. IgM or DNA were detected in 23 from 51 municipal districts of the state of Rio de Janeiro, where the samples were collected.

Evaluation of serological tests for detecting tick-borne encephalitis virus (TBEV) antibodies in animals.

Science.gov (United States)

Klaus, Christine; Beer, Martin; Saier, Regine; Schubert, Harald; Bischoff, Sabine; Süss, Jochen

2011-01-01

Tick-borne encephalitis (TBE) in animals is not well understood yet. TBE virus (TBEV) serology in several host species could be valuable for epidemiological analyses in the field as well as for the detection of clinical cases. However, performance and suitability of the available test systems are not well assessed. Therefore, we evaluated two commercial TBEV-ELISA kits in a pilot study and compared them for their suitability in veterinary applications. For this purpose, we tested 163 field collected goat sera and evaluated the results by serum neutralization test (SNT) as "gold standard". Twenty-eight SNT positive sera (17.2%) were detected. The best suited ELISA kit was used for determination of a species-specific cutoff for horses, cattle, sheep, goats, pigs, mice, dogs, rabbits and monkeys with defined sera from animals without known or with improbable contact to TBEV. The level of non-specific ELISA results does not only differ between animal species but may also be influenced by the age of the tested animals. The number of sera which tested false positive by ELISA was higher in older than in young sheep. In order to obtain defined polyclonal sera as references, two dogs, cattle, goats, sheep, rabbits and pigs each, as well as one horse and 90 mice were immunized four times with a commercially available TBEV vaccine. In conclusion, our results demonstrated that commercial TBEV-ELISA kits are suitable for application in veterinary medicine for both, verification of clinical TBE cases and epidemiological screening. However, positive ELISA results should be verified by SNT. Only a very low number of false negative ELISA-results were found.

Immunoreactive proteins of Bifidobacterium longum ssp. longum CCM 7952 and Bifidobacterium longum ssp. longum CCDM 372 identified by gnotobiotic mono-colonized mice sera, immune rabbit sera and nonimmune human sera.

Directory of Open Access Journals (Sweden)

Sabina Górska

2016-09-01

Full Text Available The Bifidobacteria show great diversity in the cell surface architecture which may influence the physicochemical properties of the bacterial cell and strain specific properties. The immunomodulatory role of bifidobacteria has been extensively studied, however studies on the immunoreactivity of their protein molecules are very limited. Here, we compared six different methods of protein isolation and purification and we report identification of immunogenic and immunoreactive protein of two human Bifidobacterium longum ssp. longum strains. We evaluated potential immunoreactive properties of proteins employing polyclonal sera obtained from germ free mouse, rabbit and human. The protein yield was isolation method-dependent and the reactivity of proteins detected by SDS-PAGE and Western blotting was heterogeneous and varied between different serum samples. The proteins with the highest immunoreactivity were isolated, purified and have them sequenced. Among the immunoreactive proteins we identified enolase, aspartokinase, pyruvate kinase, DnaK (B. longum ssp. longum CCM 7952 and sugar ABC transporter ATP-binding protein, phosphoglycerate kinase, peptidoglycan synthethase penicillin-binding protein 3, transaldolase, ribosomal proteins and glyceraldehyde 3-phosphate dehydrogenase (B. longum ssp. longum CCDM 372.

Impact of vocational interests, previous academic experience, gender and age on Situational Judgement Test performance

NARCIS (Netherlands)

Schripsema, Nienke R.; Trigt, van Anke M.; Borleffs, Jan C. C.; Cohen-Schotanus, Janke

Situational Judgement Tests (SJTs) are increasingly implemented in medical school admissions. In this paper, we investigate the effects of vocational interests, previous academic experience, gender and age on SJT performance. The SJT was part of the selection process for the Bachelor's degree

Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis

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William H Roldán

2009-05-01

Full Text Available To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis. Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%, whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.

Functional MRI of the visual cortex and visual testing in patients with previous optic neuritis

DEFF Research Database (Denmark)

Langkilde, Annika Reynberg; Frederiksen, J.L.; Rostrup, Egill

2002-01-01

to both the results of the contrast sensitivity test and to the Snellen visual acuity. Our results indicate that fMRI is a useful method for the study of ON, even in cases where the visual acuity is severely impaired. The reduction in activated volume could be explained as a reduced neuronal input......The volume of cortical activation as detected by functional magnetic resonance imaging (fMRI) in the visual cortex has previously been shown to be reduced following optic neuritis (ON). In order to understand the cause of this change, we studied the cortical activation, both the size...... of the activated area and the signal change following ON, and compared the results with results of neuroophthalmological testing. We studied nine patients with previous acute ON and 10 healthy persons served as controls using fMRI with visual stimulation. In addition to a reduced activated volume, patients showed...

Profile of NF-κBp(65/NFκBp50) among prostate specific antigen sera levels in prostatic pathologies.

Science.gov (United States)

Bouraoui, Y; Ben Jemaa, A; Rodriguez, G; Ben Rais, N; Fraile, B; Paniagua, R; Sellemi, S; Royuela, M; Oueslati, R

2012-10-01

The aim of this work was to characterise the immunoexpression of NF-κB (p50/p65) in human prostatic pathologies and to study its profiles of activation among sera prostate specific antigen antigen (PSA) according the three groups: 0-4ng/mL, 4-20ng/mL and >20ng/mL. Twenty-four men with benign prostate hyperplasia (BPH); 19 men with prostate cancer (PC) and five men with normal prostates (NP). Immunohistochemical and western blot analysis was performed. Serum levels of PSA were assayed by immulite autoanalyser. In BPH and PC samples, immunoexpressions were observed for NF-κBp65 and NF-κBp50; while in NP samples, only were detected NF-κBp50. PC samples showed immunoreactions to NF-κBp65 and NF-κBp50 more intense (respectively 24.18±0.67 and 28.23±2.01) than that observed in BPH samples (respectively18.46±2.04 and 18.66±1.59) with special localisation in the nucleus. Different profiles of NF-κBp65 immunoexpressions were observed and BPH patients with sera PSA levels between 0-4ng/mL presented a significant weak percentage compared to BPH patients with sera PSA levels between 4-20ng/mL and >20ng/mL. No immunoreactions to NF-κBp65 were observed in PC patients with sera PSA levels between 4-20ng/mL. The sensibility of both NF-κB and PSA to inflammation allowed confirming the relationship between these two molecules and its involvement in prostatic diseases progression (inflammatory and neoplasic). Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Libérer le double, la beauté sera convulsive…

OpenAIRE

Beauvoir-Dominique, Rachel

2008-01-01

Rachel Beauvoir-Dominique, Libérer le double, la beauté sera convulsive... À propos d’une collection d’art vodou. — De 1980 à ce jour, grâce au dévouement de Madame Marianne Lehmann, une collection unique de trésors du répertoire plastique vodou et « Makaya » a été rassemblée. Dotée de plusieurs milliers d’objets de culte, cette impressionnante collection n’a pas encore été exposée publiquement de manière permanente ; elle a cependant déjà attiré l’attention de nombreuses institutions nationa...

Detection of antibodies to transmissible gastroenteritis virus of swine by modified autoradiographic test

Energy Technology Data Exchange (ETDEWEB)

Stepanek, J; Hampl, J; Franz, J; Mensik, P; Skrobak, F [Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia)

1982-08-01

A modified method of autoradiographic determination of virus antibodies of gastroenteritis of swine was developed. It is based on the actual reaction between antigen bound in cells of the infected cell cultures and antibodies in tested sera, which is visualized by rabbit antibodies labelled with /sup 125/I (/sup 125/I RaSw IgG antibody) to porcine IgG, on a sensitive radiograph and evaluated by darkening at the point of positive immunological reaction. Specificity of the test and mutual comparability and reproducibility of the results were confirmed by examining the known positive and negative sera and by a comparison with the results of the virus-neutralization test. Of the 36 examined porcine blood sera, antibodies were only proved autoradiographically in the samples positive also by virus-neutralization. In experimentally infected pigs, the same dynamics of antibody production was recorded by the two tests. They were, however, demonstrated autoradiographically the eighth day after infection, while by virus neutralization test as late as 14th day. Their level increased gradually till 35th day after infection.

Multiple antibody targets on herpes B glycoproteins B and D identified by screening sera of infected rhesus macaques with peptide microarrays.

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Sven-Kevin Hotop

Full Text Available Herpes B virus (or Herpesvirus simiae or Macacine herpesvirus 1 is endemic in many populations of macaques, both in the wild and in captivity. The virus elicits only mild clinical symptoms (if any in monkeys, but can be transmitted by various routes, most commonly via bites, to humans where it causes viral encephalitis with a high mortality rate. Hence, herpes B constitutes a considerable occupational hazard for animal caretakers, veterinarians and laboratory personnel. Efforts are therefore being made to reduce the risk of zoonotic infection and to improve prognosis after accidental exposure. Among the measures envisaged are serological surveillance of monkey colonies and specific diagnosis of herpes B zoonosis against a background of antibodies recognizing the closely related human herpes simplex virus (HSV. 422 pentadecapeptides covering, in an overlapping fashion, the entire amino acid sequences of herpes B proteins gB and gD were synthesized and immobilized on glass slides. Antibodies present in monkey sera that bind to subsets of the peptide collection were detected by microserological techniques. With 42 different rhesus macaque sera, 114 individual responses to 18 different antibody target regions (ATRs were recorded, 17 of which had not been described earlier. This finding may pave the way for a peptide-based, herpes B specific serological diagnostic test.

The effect of parenteral alimentation fluid, undiluted with saline or fresh sera, on the growth of Candida albicans in vitro at 37 degrees C.

Science.gov (United States)

Lederer, T; Rippon, J; Baldwin, S; Pachman, L M

1975-02-28

Parenteral alimentation is often complicated by Candida albicans infection which may be fatal. This study investigated the effect of alimentation fluid (Aminosol) on C. albicans' growth in vitro. It was found that concentrated Aminosol (1400 millisomoles) maintained C. albicans in a viable state but inhibited replication. Dilution of alimentation fluid to physiological concentrations (300 milliosmoles) with either saline or aged pooled normal sera promoted in vitro growth of C. albicans which was equivalent to that obtained in BHI broth and was slightly less than that obtained in Sabouraud's broth. The effects of fresh sera with full complement activity were also investigated. In fresh sera appropriately diluted with physiological saline, some clumping of the yeasts was observed and all formed germ tubes. Growth as defined by budding or the formation of hyphae was inhibited. When Aminosol was diluted to 300 milliosomoles with fresh sera, all yeasts were noted to be in clumps with germ tubes as well as continually growing hyphae. Growth was approximately equal to that seen in Aminosol similarly diluted with saline.

The influence of antisperm Ig G and Ig A antibodies from cows sera and cervical mucus on bull sperm motility

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Lazarević Miodrag

2013-01-01

Full Text Available The aim of this study was to investigate the influence of antisperm Ig G and Ig A antibodies (ASA from the sera and cervical mucus of cows on bulls sperm motility. A total of 64 cows was included in the study and samples of sera and cervical mucus were collected on the day of artificial insemination. Cows were of Busha breed or mix breed with Simmental. The presence of antisperm Ig G and Ig A antibodies was determined by indirect immunofluorescence method and according to these results, cows were divided in groups as follows: cows with high or low ASA titer in their sera and cows with high or low ASA titer in the cervical mucus. Influence of antisperm antibodies on sperm motility was further estimated by Computer Assisted Semen Analysis (CASA. Results demonstrated a significant difference in the influence of antisperm antibodies depending on their origin and titer. [Projekat Ministarstva nauke Republike Srbije, br. III 46002: Molecular genetic and ecophysiological researches on the protection of autochthonous animal resources, sustaining domestic animals’ welfare, health and reproduction, and safe food production

Performance of the Epstein-Barr Virus and Herpes Simplex Virus Immunoglobulin M Assays on the Liaison Platform with Sera from Patients Displaying Acute Parvovirus B19 Infection▿

Science.gov (United States)

Costa, Elisa; Tormo, Nuria; Clari, María Ángeles; Bravo, Dayana; Muñoz-Cobo, Beatriz; Navarro, David

2009-01-01

Acute parvovirus B19 infection has been reported to cause false-positive results frequently in the Epstein-Barr (EBV) and herpes simplex virus (HSV) immunoglobulin M (IgM) assays from DiaSorin performed on the Liaison platform. We tested 65 sera from patients with a presumptive or conclusive diagnosis of acute parvovirus B19 infection in both assays and obtained no false-positive results in the EBV IgM test and 10.4% nonspecific reactivities in the HSV IgM assay. Our data support the specificity of both assays in this clinical setting. PMID:19571110

Preparation of anti-CEA and anti-goat γ-globulin sera for radioimmunologic assay of carcinoembryonic antigen

International Nuclear Information System (INIS)

Kusnierczyk-Glazman, H.; Breborowicz, J.

1977-01-01

Goats were immunized with purified carcinoembryonic antigen, and the suitability of the antisera for clinical assays of carcinoembryonic antigen was characterized. Reactivity of equine sera to goat γ-globulin as a precipitating factor in the radioimmunologic double antibody technique was also evaluated. (author)

Raman spectroscopy based screening of IgG positive and negative sera for dengue virus infection

Science.gov (United States)

Bilal, M.; Saleem, M.; Bial, Maria; Khan, Saranjam; Ullah, Rahat; Ali, Hina; Ahmed, M.; Ikram, Masroor

2017-11-01

A quantitative analysis for the screening of immunoglobulin-G (IgG) positive human sera samples is presented for the dengue virus infection. The regression model was developed using 79 samples while 20 samples were used to test the performance of the model. The R-square (r 2) value of 0.91 was found through a leave-one-sample-out cross validation method, which shows the validity of this model. This model incorporates the molecular changes associated with IgG. Molecular analysis based on regression coefficients revealed that myristic acid, coenzyme-A, alanine, arabinose, arginine, vitamin C, carotene, fumarate, galactosamine, glutamate, lactic acid, stearic acid, tryptophan and vaccenic acid are positively correlated with IgG; while amide III, collagen, proteins, fatty acids, phospholipids and fucose are negatively correlated. For blindly tested samples, an excellent agreement has been found between the model predicted, and the clinical values of IgG. The parameters, which include sensitivity, specificity, accuracy and the area under the receiver operator characteristic curve, are found to be 100%, 83.3%, 95% and 0.99, respectively, which confirms the high quality of the model.

Detection of antibodies to hepatitis B core antigen using the Abbott ARCHITECT anti-HBc assay: analysis of borderline reactive sera.

Science.gov (United States)

Ollier, Laurence; Laffont, Catherine; Kechkekian, Aurore; Doglio, Alain; Giordanengo, Valérie

2008-12-01

Routine use of the automated chemiluminescent microparticle immunoassay Abbott ARCHITECT anti-HBc for diagnosis of hepatitis B is limited in case of borderline reactive sera with low signal close to the cut-off index. In order to determine the significance of anti-HBc detection when borderline reactivity occurs using the ARCHITECT anti-HBc assay, a comparative study was designed. 3540 serum samples collected over a 2-month period in the hospital of Nice were examined for markers of HBV infection (HBsAg, anti-HBs and anti-HBc). One hundred seven samples with sufficient volume and with borderline reactivity by the ARCHITECT assay were tested by two other anti-HBc assays, a microparticle enzyme immunoassay (MEIA, AxSYM Core, Abbott Laboratories, IL, USA) and an enzyme linked fluorescent assay (ELFA, VIDAS Anti-HBc Total II, bioMérieux, Lyon, France). Only 46 samples were confirmed by the AxSYM and the VIDAS assays. Additional serological information linked to patient history showed that the remaining samples (61) were false positives (11), had low titer of anti-HBc antibodies (13), or were inconclusive (37). This comparative study highlighted the existence of a grey zone around the cut-off index. Confirmative results through a different immunoassay are needed to confirm the diagnosis of HBV on borderline reactive sera using the ARCHITECT anti-HBc assay.

21 CFR 866.3290 - Gonococcal antibody test (GAT).

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Gonococcal antibody test (GAT). 866.3290 Section... antibody test (GAT). (a) Identification. A gonococcal antibody test (GAT) is an in vitro device that..., indirect fluorescent antibody, or radioimmunoassay, antibodies to Neisseria gonorrhoeae in sera of...

Immunoreactive Proteins of Bifidobacterium longum ssp. longum CCM 7952 and Bifidobacterium longum ssp. longum CCDM 372 Identified by Gnotobiotic Mono-Colonized Mice Sera, Immune Rabbit Sera and Non-immune Human Sera.

Science.gov (United States)

Górska, Sabina; Dylus, Ewa; Rudawska, Angelika; Brzozowska, Ewa; Srutkova, Dagmar; Schwarzer, Martin; Razim, Agnieszka; Kozakova, Hana; Gamian, Andrzej

2016-01-01

The Bifidobacteria show great diversity in the cell surface architecture which may influence the physicochemical properties of the bacterial cell and strain specific properties. The immunomodulatory role of bifidobacteria has been extensively studied, however studies on the immunoreactivity of their protein molecules are very limited. Here, we compared six different methods of protein isolation and purification and we report identification of immunogenic and immunoreactive protein of two human Bifidobacterium longum ssp. longum strains. We evaluated potential immunoreactive properties of proteins employing polyclonal sera obtained from germ free mouse, rabbit and human. The protein yield was isolation method-dependent and the reactivity of proteins detected by SDS-PAGE and Western blotting was heterogeneous and varied between different serum samples. The proteins with the highest immunoreactivity were isolated, purified and have them sequenced. Among the immunoreactive proteins we identified enolase, aspartokinase, pyruvate kinase, DnaK ( B. longum ssp. longum CCM 7952) and sugar ABC transporter ATP-binding protein, phosphoglycerate kinase, peptidoglycan synthethase penicillin-binding protein 3, transaldolase, ribosomal proteins and glyceraldehyde 3-phosphate dehydrogenase ( B. longum ssp. longum CCDM 372).

Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

Directory of Open Access Journals (Sweden)

M. A. Lauricella

1993-09-01

Full Text Available This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT and hemagglutination test (IHAT were standardized, and complement fixation test (CFT and direct agglutination test (DAT were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1 parasitemic dogs of two localities of endemic area (EA of Santiago del Estero province in Argentina (n = 134; (2 non-parasitemic dogs of the same area (n = 285; (3 dogs experimentally infected with T. cruzi in the patent period (n = 6; (4 non-infected dogs (n = 56 which were born in the city of Buenos Aires (BA, one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

Culture and drug susceptibility testing among previously treated tuberculosis patients in the Dominican Republic, 2014

Directory of Open Access Journals (Sweden)

Katia J. Romero Mercado

Full Text Available ABSTRACT Multidrug-resistant tuberculosis (MDR-TB is a major public health concern that threatens global progress toward effective TB control. The risk of MDR-TB is increased in patients who have received previous TB treatment. This article describes the performance of culture and drug susceptibility testing (DST in patients registered as previously treated TB patients in the Dominican Republic in 2014, based on operational research that followed a retrospective cohort design and used routine program data. Under the current system of TB culturing and DST, the majority of patients with previously treated TB do not undergo DST, and those who do often experience considerable delay in obtaining their results. The lack of DST and delay in receiving DST results leads to underestimation of the number of MDR-TB cases and hinders the timely initiation of MDR-TB treatment.

Impact of Vocational Interests, Previous Academic Experience, Gender and Age on Situational Judgement Test Performance

Science.gov (United States)

Schripsema, Nienke R.; van Trigt, Anke M.; Borleffs, Jan C. C.; Cohen-Schotanus, Janke

2017-01-01

Situational Judgement Tests (SJTs) are increasingly implemented in medical school admissions. In this paper, we investigate the effects of vocational interests, previous academic experience, gender and age on SJT performance. The SJT was part of the selection process for the Bachelor's degree programme in Medicine at University of Groningen, the…

Impact of vocational interests, previous academic experience, gender and age on Situational Judgement Test performance.

Science.gov (United States)

Schripsema, Nienke R; van Trigt, Anke M; Borleffs, Jan C C; Cohen-Schotanus, Janke

2017-05-01

Situational Judgement Tests (SJTs) are increasingly implemented in medical school admissions. In this paper, we investigate the effects of vocational interests, previous academic experience, gender and age on SJT performance. The SJT was part of the selection process for the Bachelor's degree programme in Medicine at University of Groningen, the Netherlands. All applicants for the academic year 2015-2016 were included and had to choose between learning communities Global Health (n = 126), Sustainable Care (n = 149), Intramural Care (n = 225), or Molecular Medicine (n = 116). This choice was used as a proxy for vocational interest. In addition, all graduate-entry applicants for academic year 2015-2016 (n = 213) were included to examine the effect of previous academic experience on performance. We used MANCOVA analyses with Bonferroni post hoc multiple comparisons tests for applicant performance on a six-scenario SJT. The MANCOVA analyses showed that for all scenarios, the independent variables were significantly related to performance (Pillai's Trace: 0.02-0.47, p performance on three scenarios (p performance on two scenarios (p performance, as was previous academic experience. Gender and age were related to performance on SJT scenarios in different settings. Especially the first effect might be helpful in selecting appropriate candidates for areas of health care in which more professionals are needed.

Application of neutron activation analysis and spectrophotometry for the determination of copper level in sera and cerebrospinal fluids of schizophrenic patients

International Nuclear Information System (INIS)

Lipcsey, A.; Fekete, J.; Oerdoegh, M.; Szabo, E.

1985-01-01

Neutron activation analysis and spectrophotometry were used for the determination of copper content in sera and cerebrospinal fluids of schizophrenic patients against control persons. Comparison of the results of copper determination by both methods is tabulated. From the data the following conclusions can be drawn: for copper determinations in sera the results of the two methods agree excellently. At small copper concentrations in the cerebrospinal fluids the deviations are rather high. It can also be seen that the copper contents determined from cerebrospinal fluids taken at different times are nearly equal. (author)

Comparison of the serological tests ICT and ELISA for the diagnosis of alveolar echinococcosis in France.

Science.gov (United States)

Knapp, Jenny; Sako, Yasuhito; Grenouillet, Frédéric; Bresson-Hadni, Solange; Richou, Carine; Gbaguidi-Haore, Houssein; Ito, Akira; Millon, Laurence

2014-01-01

Serological diagnosis of alveolar echinococcosis (AE) is a key element for efficient patient treatment management. A rapid immunochromatography test kit (ICT) using the recombinant Em18 antigen (rEm18) was recently developed. The aim of our study was to assess this test on a panel of sera from French patients with alveolar echinococcosis and control patients. In a blind test, a total of 112 serum samples were tested including samples of AE (n = 30), cystic echinococcosis [CE] (n = 15), and polycystic echinococcosis [PE] (n = 1). For the comparison, 66 sera from patients with hepatocarcinoma, fascioliasis, toxocariasis, Caroli's disease, or autoimmune chronic active hepatitis were used. The diagnostic test sets we used were the rEm18-ICT and two validated ELISAs with rEm18 and Em2-Em18 antigens, respectively. For the ICT, 27/30 sera from AE patients, 4/15 sera from CE patients and the PE patient serum were positive. One serum from the control panel (toxocariasis) was positive for the ICT. The rEm18-ICT sensitivity (90.0%) and specificity (92.7%) for detection of Em18-specific antibodies confirmed it as a relevant tool for AE diagnosis. The rEm18-ELISA had a sensitivity of 86.7% and specificity of 91.5%, and the Em2-Em18-ELISA had a sensitivity of 96.7% and specificity of 87.8%. However, when AE patient sera are recorded as weak in intensity with the ICT, we recommend a double reading and use of a reference sample if the ICT is used for patient follow-up. © J. Knapp et al., published by EDP Sciences, 2014.

[Comparative study of immunoglobulins and specific antibodies in the sera of chronic brucellosis patients].

Science.gov (United States)

Kichieva, B N; Chernysheva, M I; Zheludkov, M M; Musaeva, N B

1982-04-01

The data on the IgA, IgM and IgG levels in the sera of 89 patients with chronic brucellosis lasting for 1-10 years and longer are presented. The chronic form of brucellosis is characterized by the normal or low level of immunoglobulins. No correlation between the levels of IgG, IgM and the titer of specific antibodies has been established.

Precision of glucose measurements in control sera by isotope dilution/mass spectrometry: proposed definitive method compared with a reference method

International Nuclear Information System (INIS)

Pelletier, O.; Arratoon, C.

1987-01-01

This improved isotope-dilution gas chromatographic/mass spectrometric (GC/MS) method, in which [ 13 C]glucose is the internal standard, meets the requirements of a Definitive Method. In a first study with five reconstituted lyophilized sera, a nested analysis of variance of GC/MS values indicated considerable among-vial variation. The CV for 32 measurements per serum ranged from 0.5 to 0.9%. However, concentration and uncertainty values (mmol/L per gram of serum) assigned to one serum by the NBS Definitive Method (7.56 +/- 0.28) were practically identical to those obtained with the proposed method (7.57 +/- 0.20). In the second study, we used twice more [ 13 C]glucose diluent to assay four serum pools and two lyophilized sera. The CV ranged from 0.26 to 0.5% for the serum pools and from 0.28 to 0.59% for the lyophilized sera. In comparison, results by the hexokinase/glucose-6-phosphate dehydrogenase reference method agreed within acceptable limits with those by the Definitive Method but tended to be slightly higher (up to 3%) for lyophilized serum samples or slightly lower (up to 2.5%) for serum pools

21 CFR 864.9300 - Automated Coombs test systems.

Science.gov (United States)

2010-04-01

... Blood and Blood Products § 864.9300 Automated Coombs test systems. (a) Identification. An automated Coombs test system is a device used to detect and identify antibodies in patient sera or antibodies bound... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated Coombs test systems. 864.9300 Section...

Indirect micro-immunofluorescence test for detecting type-specific antibodies to herpes simplex virus.

Science.gov (United States)

Forsey, T; Darougar, S

1980-02-01

A rapid indirect micro-immunofluorescence test capable of detecting and differentiating type-specific antibodies to herpes simplex virus is described. The test proved highly sensitive and, in 80 patients with active herpes ocular infection, antibody was detected in 94%. No anti-herpes antibody was detected in a control group of 20 patients with adenovirus infections. Testing of animal sera prepared against herpes simplex virus types 1 and 2 and of human sera from cases of ocular and genital herpes infections showed that the test can differentiate antibodies to the infecting serotypes. Specimens of whole blood, taken by fingerprick, and eye secretions, both collected on cellulose sponges, could be tested by indirect micro-immunofluorescence. Anti-herpes IgG, IgM, and IgA can also be detected.

[Comparison of eight screening tests for ant-HCV antibody].

Science.gov (United States)

Deguchi, Matsuo; Kagita, Masanori; Yamashita, Naoko; Nakano, Takasi; Tahara, Kazuko; Asari, Seishi; Iwatani, Yoshinori

2002-09-01

We compared eight HCV screening tests for detection of anti-HCV antibody; Ortho Quick Chaser HCV Ab (QC), Ortho HCV Ab ELISA III (ELISA), Ortho HVC Ab PA test III (PA), Lumipulse II Ortho HCV (LUMI), IMx HCV.DAINAPACKII (IMx), ARCHITECT HCV (ARCH), Immucheck.F-HCV C50 Ab (Immu), RANREAM HCV Ab Ex II (RAN). Sera from six hundred patients were examined by these eight screening tests. The positive rates of the eight screening tests were from 9.0% to 13.2%. Forty-five sera showed discrepant results between the eight screening tests, and about half of them showed weak positive reaction and/or false positive. Twenty-five of the forty-five sera were negative for ant-HCV antibody in the CHIRON RIBA III confirmatory test, and forty-four of them were negative for HCV-RNA in the PCR method. The agreement rates between the two reagents were from 95.5% to 99.2%, but were not always high between the two reagents that used similar antigen. The specificities and sensitivities evaluated by using the RIBA III confirmatory test were excellent in ELISA, LUMI, IMx, ARCH and Immu. Three BBI seroconversion panels were used to compare the positive readings in the initial stage of HCV infection by eight screening tests. ELISA and ARCH showed the earliest positive readings, and then IMx, LUMI = RAN, PA, QC and Immu in this order. These findings indicate that ELISA and ARCH were the most excellent in the sensitivity, specificity and early diagnosis of HCV infection. However, we must pay attention to the weak positive reaction in the screening tests, because there is a possibility of "false positive".

Immunochemical characterization of Mycobacterium leprae antigens by the SDS-polyacrylamide gel electrophoresis immunoperoxidase technique (SGIP) using patients' sera

NARCIS (Netherlands)

Klatser, P. R.; van Rens, M. M.; Eggelte, T. A.

1984-01-01

In this study the SDS-polyacrylamide gel electrophoresis immunoperoxidase (SGIP) assay was used for characterizing the antigenic components of Mycobacterium leprae using patients' sera. This technique involved the separation of mycobacterial sonicates on SDS-polyacrylamide gels, longitudinal

Electrophoretic pattern of sera from lambs and kids vaccinated with irradiated Amphistome metacercariae (Cercariae indicae XXVI)

International Nuclear Information System (INIS)

Hafeez, Md.; Rao, B.V.

1986-01-01

Preliminary work has been done to study certain responses induced by irradiated amphistome metacercariae used as a vaccine to immunise lambs, kids and calves. The electrophoretic pattern of the sera collected from lambs and kids vaccinated with gamma irradiated amphistome matacercariae (C.I. XXVI) has been reported in this study. (author). 10 refs., 1 table

Sensitivity and specificity of various serologic tests for detection of Toxoplasma gondii infection in naturally infected sows

DEFF Research Database (Denmark)

Dubey, J.P.; Thulliez, P.; Weigel, R.M.

1995-01-01

antibodies by use of the modified agglutination test (MAT), latex agglutination test (LAT), indirect hemagglutination test (IHAT), and ELISA. Toxoplasma gondii was isolated from 170 hearts of 1,000 sows by bioassays in mice and cats. The percentage of samples diagnosed as positive for each of the serologic......, 29.4 and 98.3% for IHAT, 45.9 and 96.9% for LAT, and 72.9 and 85.9% for ELISA. The dye test was run at 1:20 dilution on only 893 sera because of bacterial contamination and presence of anticomplement substances. Dye test antibodies were found in 17.8% of the sera, and sensitivity and specificity were...

Quantitation of parasite-specific human IgG and IgE in sera: evaluation of solid-phase RIA and ELISA methodology

Energy Technology Data Exchange (ETDEWEB)

Hamilton, R G [Johns Hopkins Univ., Baltimore, MD (USA). Dept. of Medicine; Hussain, R; Ottesen, E A [National Inst. of Allergy and Infectious Diseases, Bethesda, MD (USA); Adkinson, Jr, N F [Johns Hopkins Univ., Baltimore, MD (USA). School of Medicine

1981-07-17

The authors have developed a non-competitive solid-phase radioimmunoassay (SPRIA) to quantitate both human IgE and IgG antibodies against soluble adult antigens of Brugia malayi (B.m.), a filarial parasite causing extensive infection throughout the tropics. Previously enzyme-linked immunosorbent assays (ELISA) had been used to detect ..mu..g/ml levels of IgG anti-B.m., but IgE antibodies were difficult to detect in this system. Since the SPRIA successfully quantitates both IgG and IgE anti-B.m., they sought to examine the reasons for the SPRIA's apparent superiority in detecting IgE anti-B.m. by extracting specific IgG from sera with high levels of IgE and IgG anti-B.m. antibodies. IgE anti-B.m. was then quantitated in these sera using both the SPRIA and ELISA methods. Results indicate that IgG anti-B.m. does not interfere with detection of specific IgE antibody in the SPRIA but does interfere in the ELISA. While ELISA permits detection of IgE anti-B.m. in the absence of competing IgG anti-B.m., as levels of specific IgG increase, the IgE is no longer detectable. These differences between SPRIA and ELISA can be explained by the SPRIA's antigen excess conditions which assure that there are sufficient antigens both to detect all anti-B.m. antibodies present in the serum and to adequately represent all antigen specificities in the crude B.m. extract. Their findings commend the use of SPRIA methods over ELISA in assessment of B.m.-specific IgE antibody in filariasis and indicate a potential role for SPRIA methods in absolute quantitation of specific serum antibodies.

Assessing the validity of an ELISA test for the serological diagnosis of human fascioliasis in different epidemiological situations.

Science.gov (United States)

Valero, M Adela; Periago, M Victoria; Pérez-Crespo, Ignacio; Rodríguez, Esperanza; Perteguer, M Jesús; Gárate, Teresa; González-Barberá, Eva M; Mas-Coma, Santiago

2012-05-01

To improve the diagnosis of human fascioliasis caused by Fasciola hepatica and Fasciola gigantica, we evaluated the diagnostic accuracy of an enzyme-linked immunosorbent assay (ELISA), with Fasciola antigen from the adult liver fluke, for the detection of IgG against fascioliasis in human sera. The sera of 54 fascioliasis cases, originating from three endemic areas, were used in this evaluation: (i) a hyperendemic F. hepatica area where humans usually shed a great number of parasite eggs in faeces (11 sera); (ii) an epidemic F. hepatica area where humans usually shed small amounts of parasite eggs (24 sera) and (iii) an overlap area of both Fasciola species and where human shedding of parasite eggs in faeces is usually scarce or non-existent (19 sera). One hundred and sixty-eight patients with other parasitic infections and 89 healthy controls were also analysed. The respective sensitivity and specificity of this assay were 95.3% (95% confidence intervals, 82.9-99.2%) and 95.7% (95% confidence intervals, 92.3-97.5%). No correlation between egg output and the OD450 values of the F. hepatica IgG ELISA test was observed. This test could be used both as an individual serodiagnostic test for human fascioliasis when backed up by a compatible clinical history together with a second diagnostic technique for other cross-reactive helminth infections, and in large-scale epidemiological studies of human fascioliasis worldwide. © 2012 Blackwell Publishing Ltd.

Apoptosis induction in Jurkat cells and sCD95 levels in women's sera are related with the risk of developing cervical cancer

International Nuclear Information System (INIS)

Aguilar-Lemarroy, Adriana; Jave-Suarez, Luis F; Romero-Ramos, Jose E; Olimon-Andalon, Vicente; Hernandez-Flores, Georgina; Lerma-Diaz, Jose M; Ortiz-Lazareno, Pablo C; Morgan-Villela, Gilberto; Toro-Arreola, Susana del; Bravo-Cuellar, Alejandro

2008-01-01

Currently, there is clear evidence that apoptosis plays an important role in the development and progression of tumors. One of the best characterized apoptosis triggering systems is the CD95/Fas/APO-1 pathway; previous reports have demonstrated high levels of soluble CD95 (sCD95) in serum of patients with some types of cancer. Cervical cancer is the second most common cancer among women worldwide. As a first step in an attempt to design a minimally invasive test to predict the risk of developing cervical cancer in patients with precancerous lesions, we used a simple assay based on the capacity of human serum to induce apoptosis in Jurkat cells. We evaluated the relationship between sCD95 levels and the ability to induce apoptosis in Jurkat cells in cervical cancer patients and controls. Jurkat cells were exposed to serum from 63 women (20 healthy volunteers, 21 with cervical intraepithelial neoplasia grade I [CIN 1] and 22 with cervical-uterine carcinoma). The apoptotic rate was measured by flow cytometry using Annexin-V-Fluos and Propidium Iodide as markers. Serum levels of sCD95 and soluble CD95 ligand (sCD95L) were measured by ELISA kits. We found that serum from almost all healthy women induced apoptosis in Jurkat cells, while only fifty percent of the sera from women with CIN 1 induced cell death in Jurkat cells. Interestingly, only one serum sample from a patient with cervical-uterine cancer was able to induce apoptosis, the rest of the sera protected Jurkat cells from this killing. We were able to demonstrate that elimination of Jurkat cells was mediated by the CD95/Fas/Apo-1 apoptotic pathway. Furthermore, the serum levels of sCD95 measured by ELISA were significantly higher in women with cervical cancer. Our results demonstrate that there is a strong correlation between low levels of sCD95 in serum of normal women and higher apoptosis induction in Jurkat cells. We suggest that an analysis of the apoptotic rate induced by serum in Jurkat cells and the

Evaluation of three commercial rapid tests for detecting antibodies to human immunodeficiency virus.

Science.gov (United States)

Ng, K P; Saw, T L; Baki, A; Kamarudin, R

2003-08-01

Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest are simple/rapid tests for the detection of antibodies to HIV-1 and HIV-2 in human whole blood, serum and plasma samples. The assay is one step and the result is read visually within 15 minutes. Using 92 known HIV-1 reactive sera and 108 known HIV-1 negative sera, the 3 HIV tests correctly identified all the known HIV-1 reactive and negative samples. The results indicated that Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest HIV are as sensitive and specific (100% concordance) as Microparticle Enzyme Immunoassay. The data indicated that these 3 HIV tests are effective testing systems for diagnosis of HIV infection in a situation when the conventional Enzyme Immunoassay is not suitable.

Comparative thermal cyclic test of different beryllium grades previously subjected to simulated disruption loads

International Nuclear Information System (INIS)

Gervash, A.; Giniyatulin, R.; Mazul, I.

1999-01-01

Considering beryllium as plasma facing armour this paper presents recent results obtained in Russia. A special process of joining beryllium to a Cu-alloy material structure is described and recent results of thermal cycling tests of such joints are presented. Summarizing the results, the authors show that a Cu-alloy heat sink structure armoured with beryllium can survive high heat fluxes (≥10 MW/m 2 ) during 1000 heating/cooling cycles without serious damage to the armour material and its joint. The principal feasibility of thermal cycling of beryllium grades and their joints directly in the core of a nuclear reactor is demonstrated and the main results of this test are presented. The paper also describes the thermal cycling of different beryllium grades having cracks initiated by previously applied high heat loads simulating plasma disruptions. (orig.)

Quantitation of IgE by means of a modified radial immunodiffusion method in comparison with the radioimmunosorbent test (RIST)

International Nuclear Information System (INIS)

Wiedermann, G.; Stemberg, H.; Kraft, D.; Ambrosch, F.; Schadlbauer, B.; Jarischko, E.; Vienna Univ.

1974-01-01

Serum IgE were quantitied by means of modified radial immunodiffusion technique (RID). To improve visibility of precipitin bands a staining procedure with DOPA was applied. Pretreatment of sera with dextransulfate proved necessary in order to avoid unspecific ringformation in the agargel. In comparison with the RIST it turned out that sera containing less than 500 I.U. IgE/ml did not produce precipitin bands with this method. Sera containing 500-999 I.U. IgE/ml occasionally exhibited positive results with the RID technique, whereas sera with more than 1,000 I.U./ml were regulary positive. In its present form the RID may be used as screening method for sera with higher IgE levels. Within the above mentioned limits the IgE levels calculated by means of the RID-test roughly corresponded the values determined by the RIST. (orig.) [de

IgG western blot for confirmatory diagnosis of equivocal cases of toxoplasmosis by EIA-IgG and fluorescent antibody test.

Science.gov (United States)

Khammari, Imen; Saghrouni, Fatma; Yaacoub, Alia; Gaied Meksi, Sondoss; Ach, Hinda; Garma, Lamia; Fathallah, Akila; Ben Saïd, Moncef

2013-08-01

The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.

Mass spectrometry data from proteomics-based screening of immunoreactive proteins of fully virulent Brucella strains using sera from naturally infected animals

Directory of Open Access Journals (Sweden)

Gamal Wareth

2015-09-01

Full Text Available Here, we provide the dataset associated with our research article on comprehensive screening of Brucella immunoreactive proteins using sera of naturally infected hosts published in Biochemical and Biophysical Research Communications Wareth et al., 2015 [1]. Whole-cell protein extracts were prepared from Brucella abortus and Brucella melitensis, separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE and subsequently western blotting was carried out using sera from bovines (cows and buffaloes and small ruminants (goats and sheep. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org via the PRIDE partner repository [2] with the dataset identifiers PXD001270 and DOI:10.6019/PXD001270.

Recombinant major outer membrane protein (MOMP) of Chlamydophila abortus, Chlamydophila pecorum, and Chlamydia suis as antigens to distinguish chlamydial species-specific antibodies in animal sera.

Science.gov (United States)

Hoelzle, Ludwig E; Hoelzle, Katharina; Wittenbrink, Max M

2004-10-05

Recombinant major outer membrane proteins (rMOMP) of Chlamydophila (Ch.) abortus, Ch. pecorum, and Chlamydia (C.) suis were used as antigens to distinguish chlamydial species-specific antibodies in (i) immune sera from six rabbits and three pigs raised against native purified elementary bodies, (ii) serum samples from 25 sows vaccinated with Ch. abortus, and (iii) 40 serum samples from four heifers experimentally infected with Ch. abortus. All post-exposition sera contained chlamydial antibodies as confirmed by strong ELISA seroreactivities against the chlamydial LPS. For the rMOMP ELISA mean IgG antibody levels were at least 5.8-fold higher with the particular rMOMP homologous to the chlamydial species used for immunisation or infection than with heterologous rMOMPs (P <0.001). Preferential rMOMP ELISA reactivities of sera were confirmed by Western blotting. The results suggest that the entire chlamydial rMOMP could provide a species-specific serodiagnostic antigen.

Reliability and smallest worthwhile difference in 1RM tests according to previous resistance training experience in young women

Directory of Open Access Journals (Sweden)

Matheus Amarante do Nascimento

2017-10-01

Full Text Available The objective of this study was to determine the familiarization and smallest worthwhile difference (SWD of one-repetition maximum (1RM tests in detrained women according to their previous resistance training experience. Three groups of women with varying amounts of previous resistance training experience were recruited: Novice (n = 27, 1 to 6 months, Intermediate (n = 13, from 7 to 12 months, and Advanced (n = 20, 13 to 24 months. All participants performed four 1RM test sessions in the bench press (BP, squat (SQ, and arm curl (AC. A significant (p< 0.05 (group vs. time interaction was observed in SQ suggesting that more experienced participants needed fewer 1RM test sessions to reach a stable load compared to the less experienced groups. Strength changes (p 0.05, suggesting that experience had no impact on familiarization for these lifts. SWDs suggest that strength gains greater than 2-4% in these lifts would indicate a meaningful improvement in strength beyond random variation from trial to trial no matter the experience of the subject. Women with limited previous resistance training experience do not require more trials to reach load stabilization than those with more experience. Stability of 1RM loads for BP and AC may require only two sessions, while SQ may require at least three trials.

Detection of a novel specificity (CTLA-4) in ATG/TMG globulins and sera from ATG-treated leukemic patients.

Science.gov (United States)

Pistillo, Maria Pia; Tazzari, Pier Luigi; Bonifazi, Francesca; Bandini, Giuseppe; Kato, Tomohiro; Matsui, Toshihiro; Nishioka, Kusuki; Conte, Roberto; Ferrara, Giovanni Battista

2002-04-27

T-cell costimulation has been shown to provide positive signals for T-cell activation and generation of effector activity. In this study, we analyzed the presence of antibodies (Abs) against the T-lymphocyte costimulatory molecules CD28, CTLA-4, CD80, and CD86 in anti-T-lymphocyte (ATG) and antithymocyte (TMG) globulin preparations to address their mechanism of action. We focused our attention on the role of CTLA-4-specific Abs in the immunosuppressive effect of ATG/TMG, because anti-CTLA-4 agonistic Abs may suppress T-cell proliferation and nonagonistic Abs may lead to T-cell depletion through an Ab-dependent cell cytotoxicity mechanism. ATG/TMG and patients' sera were tested for binding to recombinant human costimulatory molecules by ELISA techniques. CTLA-4 specificity was also analyzed by cytoplasmic immunofluorescence staining of a CTLA-4 transfectant by competitive inhibition immunofluorescence and by cell proliferation assay in allogeneic mixed lymphocyte reaction (MLR). Either ATG or TMG predominantly contained anti-CTLA-4 Abs, with higher reactivity in ATG followed by anti-CD86 and -CD28 Abs, whereas anti-CD80 Abs were found only in ATG. Anti-CTLA-4 Abs present in ATG/TMG recognized the native form of CTLA-4 molecule, and their removal reduced the effect of ATG in an allogeneic MLR. Kinetic studies indicated that such Abs were present in the sera of 12 ATG-treated leukemic patients up to 21 days after ATG administration. These data suggest that the novel anti-CTLA-4 Abs found in ATG may greatly contribute to its immunosuppressive effect, thus accounting for the absence of rejection and exceptionally low incidence of graft-versus-host disease in the group of patients analyzed.

[IgM class antibodies against hepatitis A viruses in the differentiation of viral hepatitis].

Science.gov (United States)

Zivanović-Marinković, V; Stanković, D; Parabucki, S; Lazarov, A; Marinković, V; Krstić, L; Letica, Z

1982-01-01

The sera of 64 patients with acute viral hepatitis which were previously HbsAG negative by rPHA were tested by RIA methods for the presence of HBsAG and anti-HBc, by EIA method for HBc and anti-HBc and also by RIA method for the presence of IgM antibodies against hepatitis A virus. The pairs of sera were tested also for the presence of antibodies against cytomegalovirus and EB viruses. Of the total of 64 patients, markers of fresh HB viral infection were proved in 5 patients, sera positive to IgM antibodies to hepatitis A virus were found in 51 and no positive tests to any of the tested viruses were found in 8. The authors consider that they belonged to "non-A, non-B" viral hepatitis.

A large portion of meningococcal antigen typing system-negative meningococcal strains from spain is killed by sera from adolescents and infants immunized with 4CMenB.

Science.gov (United States)

Abad, R; Biolchi, A; Moschioni, M; Giuliani, M M; Pizza, M; Vázquez, J A

2015-04-01

A new vaccine (the 4CMenB 4-component protein vaccine [Bexsero], which includes PorA, factor H-binding protein [fHbp], neisserial heparin-binding antigen [NHBA], and Neisseria adhesin A [NadA]) against serogroup B meningococci has recently been approved for use in people older than age 2 months in Europe, Australia, and Canada. Preapproval clinical efficacy studies are not feasible for invasive meningococcal disease because its incidence is low/very low, and the serum bactericidal antibody (SBA) titer (or the human SBA [hSBA] titer when human complement is used in the assay) has been used as a surrogate marker of protection. However, the hSBA assay cannot be used on a large scale, and therefore, a meningococcal antigen typing system (MATS) was developed. MATS combines conventional PorA genotyping with an enzyme-linked immunosorbent assay (ELISA) that quantifies both the expression and the cross-reactivity of antigenic variants. The assay has been used to evaluate the potential of the 4CMenB meningococcal group B vaccine to cover group B strains in several countries. Some recent data suggest that MATS is a conservative predictor of strain coverage. We used pooled sera from adolescents and infants to test by the hSBA assay 10 meningococcal group B strains isolated in Spain that were negative for the 3 antigens (n = 9) or that had very low levels of the 3 antigens (n = 1) by MATS. We found that all strains were killed by sera from adolescents and that 5 of the 10 strains were also killed, although at a low titer, by sera from infants. Our data confirm that MATS underestimates vaccine coverage. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Apoptosis induction in Jurkat cells and sCD95 levels in women's sera are related with the risk of developing cervical cancer

Directory of Open Access Journals (Sweden)

Bravo-Cuellar Alejandro

2008-04-01

Full Text Available Abstract Background Currently, there is clear evidence that apoptosis plays an important role in the development and progression of tumors. One of the best characterized apoptosis triggering systems is the CD95/Fas/APO-1 pathway; previous reports have demonstrated high levels of soluble CD95 (sCD95 in serum of patients with some types of cancer. Cervical cancer is the second most common cancer among women worldwide. As a first step in an attempt to design a minimally invasive test to predict the risk of developing cervical cancer in patients with precancerous lesions, we used a simple assay based on the capacity of human serum to induce apoptosis in Jurkat cells. We evaluated the relationship between sCD95 levels and the ability to induce apoptosis in Jurkat cells in cervical cancer patients and controls. Methods Jurkat cells were exposed to serum from 63 women (20 healthy volunteers, 21 with cervical intraepithelial neoplasia grade I [CIN 1] and 22 with cervical-uterine carcinoma. The apoptotic rate was measured by flow cytometry using Annexin-V-Fluos and Propidium Iodide as markers. Serum levels of sCD95 and soluble CD95 ligand (sCD95L were measured by ELISA kits. Results We found that serum from almost all healthy women induced apoptosis in Jurkat cells, while only fifty percent of the sera from women with CIN 1 induced cell death in Jurkat cells. Interestingly, only one serum sample from a patient with cervical-uterine cancer was able to induce apoptosis, the rest of the sera protected Jurkat cells from this killing. We were able to demonstrate that elimination of Jurkat cells was mediated by the CD95/Fas/Apo-1 apoptotic pathway. Furthermore, the serum levels of sCD95 measured by ELISA were significantly higher in women with cervical cancer. Conclusion Our results demonstrate that there is a strong correlation between low levels of sCD95 in serum of normal women and higher apoptosis induction in Jurkat cells. We suggest that an analysis of

The quantitation of parasite-specific human IgG and IgE in sera: evaluation of solid-phase RIA and ELISA methodology

International Nuclear Information System (INIS)

Hamilton, R.G.; Adkinson, N.F. Jr.

1981-01-01

The authors have developed a non-competitive solid-phase radioimmunoassay (SPRIA) to quantitate both human IgE and IgG antibodies against soluble adult antigens of Brugia malayi (B.m.), a filarial parasite causing extensive infection throughout the tropics. Previously enzyme-linked immunosorbent assays (ELISA) had been used to detect μg/ml levels of IgG anti-B.m., but IgE antibodies were difficult to detect in this system. Since the SPRIA successfully quantitates both IgG and IgE anti-B.m., they sought to examine the reasons for the SPRIA's apparent superiority in detecting IgE anti-B.m. by extracting specific IgG from sera with high levels of IgE and IgG anti-B.m. antibodies. IgE anti-B.m. was then quantitated in these sera using both the SPRIA and ELISA methods. Results indicate that IgG anti-B.m. does not interfere with detection of specific IgE antibody in the SPRIA but does interfere in the ELISA. While ELISA permits detection of IgE anti-B.m. in the absence of competing IgG anti-B.m., as levels of specific IgG increase, the IgE is no longer detectable. These differences between SPRIA and ELISA can be explained by the SPRIA's antigen excess conditions which assure that there are sufficient antigens both to detect all anti-B.m. antibodies present in the serum and to adequately represent all antigen specificities in the crude B.m. extract. Their findings commend the use of SPRIA methods over ELISA in assessment of B.m.-specific IgE antibody in filariasis and indicate a potential role for SPRIA methods in absolute quantitation of specific serum antibodies. (Auth.)

Persistence of atovaquone in human sera following treatment: inhibition of Plasmodium falciparum development in vivo and in vitro.

Science.gov (United States)

Butcher, Geoff A; Sinden, Robert E

2003-01-01

Published pharmacokinetic data indicate that after treatment of patients with therapeutic doses of atovaquone/proguanil hydrochloride (Malarone, GlaxoSmithKline Research Triangle Park, NC), the plasma half-lives of these drugs are 70h and 15h, respectively. However, using two biologic assays (mosquito transmission and in vitro asexual stage development), we demonstrate here that sera from volunteers treated with atovaquone/proguanil retained activity against Plasmodium falciparum up to 6 weeks after such treatment. This activity was due to atovaquone, as administration of this drug alone replicated the data obtained with the combination. Most notably, asexual stage development of an atovaquone-resistant strain (NGATV01) of P. falciparum was not inhibited by sera taken after atovaquone treatment. These data indicate that for atovaquone, biologic assays, though not quantitative, are more sensitive than the usual physicochemical assays. Also, persistence of atovaquone in plasma at low concentrations for long periods may increase the risk of resistant parasites arising.

Immunoreactivity of protein tyrosine phosphatase A (PtpA) in sera from sheep infected with Mycobacterium avium subspecies paratuberculosis.

Science.gov (United States)

Gurung, Ratna B; Begg, Douglas J; Purdie, Auriol C; Bach, Horacio; Whittington, Richard J

2014-07-15

Evasion of host defense mechanisms and survival inside infected host macrophages are features of pathogenic mycobacteria including Mycobacterium avium subspecies paratuberculosis, the causative agent of Johne's disease in ruminants. Protein tyrosine phosphatase A (PtpA) has been identified as a secreted protein critical for survival of mycobacteria within infected macrophages. The host may mount an immune response to such secreted proteins. In this study, the humoral immune response to purified recombinant M. avium subsp. paratuberculosis PtpA was investigated using sera from a cohort of sheep infected with M. avium subsp. paratuberculosis and compared with uninfected healthy controls. A significantly higher level of reactivity to PtpA was observed in sera collected from M. avium subspecies paratuberculosis infected sheep when compared to those from uninfected healthy controls. PtpA could be a potential candidate antigen for detection of humoral immune responses in sheep infected with M. avium subspecies paratuberculosis. Copyright © 2014 Elsevier B.V. All rights reserved.

Rapid differentiation of rocky mountain spotted fever from chickenpox, measles, and enterovirus infections and bacterial meningitis by frequency-pulsed electron capture gas-liquid chromatographic analysis of sera.

Science.gov (United States)

Brooks, J B; McDade, J E; Alley, C C

1981-01-01

Normal sera and sera from patients with Rocky Mountain spotted fever, chickenpox, enterovirus infections, measles, and Neisseria meningitidis infections were extracted with organic solvents under acidic and basic conditions and then derivatized with trichloroethanol or heptafluorobutyric anhydride-ethanol to form electron-capturing derivatives of organic acids, alcohols, and amines. The derivatives were analyzed by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). There were unique differences in the FPEC-GLC profiles of sera obtained from patients with these respective diseases. With Rocky Mountain spotted fever patients, typical profiles were detected as early as 1 day after onset of disease and before antibody could be detected in the serum. Rapid diagnosis of Rocky Mountain spotted fever by FPEC-GLC could permit early and effective therapy, thus preventing many deaths from this disease. PMID:7276147

[Comparison of conventional and non-conventional serological tests for the diagnosis of imported Chagas disease in Spain].

Science.gov (United States)

Flores-Chávez, María; Cruz, Israel; Rodríguez, Mercedes; Nieto, Javier; Franco, Elena; Gárate, Teresa; Cañavate, Carmen

2010-05-01

Trypanosoma cruzi infection is a major imported parasitic disease in Spain, because of the increase of immigrants from endemic areas. Since the laboratory diagnosis during the chronic phase is based on detection of anti-T. cruzi IgG antibodies, our aims were to compare 10 tests for determining anti-T. cruzi antibodies, to assess their cross-reactivity with related diseases, and to evaluate the rk39-ELISA and IFAT-Leishmania tests as tools for the differential diagnosis of leishmaniasis due to Leishmania infantum. A total of 223 sera were tested: 40 had been previously characterized by Qpanel, and 183 were obtained from the serum library of the Parasitology Department, Centro Nacional de Microbiología (66 chagasic, 97 healthy, 30 visceral leishmaniasis, and 30 malaria). Samples were examined using in-house IFAT and ELISA, 5 commercial ELISAs (Certest/Abbot Laboratories/BiosChile; Ortho Clinical Diagnostics; BLK Diagnostic; bioMérieux; and Biokit), particle gel agglutination (ID-PaGIA), and two immunochromatographic assays (Operon and CTK Biotech). The last 4 tests are based in recombinant antigens (non-conventional tests). The IFAT and ELISAs showed a sensitivity of 97% to 100%. The immunochromatographic tests had somewhat lower sensitivity (92%-96%). All non-conventional tests presented a smaller number of cross-reactions. Leishmania-Rk39-ELISA did not show cross-reactivity with chagasic sera. In general, our results confirm the data obtained by other authors. The sensitivity of ELISA is higher than other tests; therefore, these techniques would be the most appropriate for screening of T. cruzi infection. A suitable approach is the combination of a test using total antigen with another based on either recombinant antigens or synthetic peptides. (c) 2009 Elsevier España, S.L. All rights reserved.

Discrepancies in Weil-Felix and microimmunofluorescence test results for Rocky Mountain spotted fever.

Science.gov (United States)

Hechemy, K E; Stevens, R W; Sasowski, S; Michaelson, E E; Casper, E A; Philip, R N

1979-01-01

Only 4.2% of 284 single specimens and 17.6% of 51 pairs of sera reactive in Weil-Felix agglutination tests for Rocky Mountain spotted fever were confirmed by a specific Rickettsia rickettsii microimmunofluorescence test. PMID:107194

Determine Multiple Elements Simultaneously in the Sera of Umbilical Cord Blood Samples-a Very Simple Method.

Science.gov (United States)

Liang, Chunmei; Li, Zhijuan; Xia, Xun; Wang, Qunan; Tao, Ruiwen; Tao, Yiran; Xiang, Haiyun; Tong, Shilu; Tao, Fangbiao

2017-05-01

Analyzing the concentrations of heavy metals in the sera of umbilical cord blood samples can provide useful information about prenatal exposure to environmental agents. An analytical method based on ICP-MS to simultaneously determine multiple elements in umbilical cord blood samples was developed for assessing the in utero exposure to metallic and metalloid elements. The method only required as little as 100 μL of serum diluted 1:25 for direct analysis. Matrix-matched protocol was used to eliminate mass matrix interference and kinetic energy discrimination mode was used to eliminate the polyatomic ion interference. The assay was completed on average within 4 min with the detection limits ranging from 0.0002 to 44.4 μg/L for all the targeted elements. The detection rates for most of elements were 100 % other than cadmium (Cd), lead (Pb), and mercury (Hg). The testing results of the certified reference materials were ideal. The method is simple and sensitive, so it is suitable for the monitoring of large quantities of samples.

Thyroxine determination in serum by radioimmunoassay and competitive protein-binding analysis (simultaneous study using four test kits)

International Nuclear Information System (INIS)

Wagner, K.

1978-01-01

Three commercial test kits for radioimmunological T 4 -determination have been tested (T 4 -RIA Abbott, RIA-mat T 4 , T 4 -RIA Henning). The RIA method has also been compared with a CPBA test (Tetralute) for routine application. 1) The results show that CPBA (Tetralute) is superior over the three radioimmuno-assays with regard to intra-assay variance. The T 4 -RIA Abbott has been inferior to the two other RIA under test and to the Tetralute with regard to inter-assay variance. 2) The four test kits show significant differences and considerable deviations from the given nominal values in comparison with the mean values of two control sera. A control for correctness does not seem to be appropriate as it cannot be decided which of the test kits yields the true value of T 4 content. No systematic difference has been observed between RIA and CPBA in the determination of T 4 in control sera. 3) No detectable difference has been found among the RIA-T 4 test kits and comparing them with the CBPA test regarding their diagnostic precision. A good correlation of T 4 values of euthyroid patients comparing the Tetralute and the three radioimmunological test kits. The mean values of euthyroid sera obtained by the three radioimmuno-assays and the Tetralute are not in agreement with each other. (orig.) [de

Increased Expression of Cytotoxic T-Lymphocyte-Associated Protein 4 by T Cells, Induced by B7 in Sera, Reduces Adaptive Immunity in Patients With Acute Liver Failure

DEFF Research Database (Denmark)

Khamri, Wafa; Abeles, Robin D; Hou, Tie Zheng

2017-01-01

, hepatic sinusoidal endothelial cells, and biliary epithelial cells from healthy or diseased liver tissues. We also measured levels of soluble B7 serum samples from patients and controls, and mice with acetaminophen-induced liver injury using enzyme-linked immunosorbent assays. RESULTS: Peripheral blood...... were found to have increased concentrations of soluble B7 compared to sera from controls. Necrotic human primary hepatocytes exposed to acetaminophen, but not hepatic sinusoidal endothelial cells and biliary epithelial cells from patients with ALF, secreted high levels of soluble B7. Sera from mice...... with acetaminophen-induced liver injury contained high levels of soluble B7 compared to sera from mice without liver injury. Plasma exchange reduced circulating levels of soluble B7 in patients with ALF and expression of CTLA4 on T cells. CONCLUSIONS: Peripheral CD4+T cells from patients with ALF have increased...

Comparison of a new rapid plasma reagin card test with the standard rapid plasma reagin 18-mm circle card test and the venereal disease research laboratory slide test for serodiagnosis of syphilis.

OpenAIRE

Hambie, E A; Larsen, S A; Perryman, M W; Pettit, D E; Mullally, R L; Whittington, W

1983-01-01

The rapid plasma reagin (RPR) card test manufactured by Beckman Instruments, Inc., was compared, qualitatively and quantitatively, with the Venereal Disease Research Laboratory (VDRL) slide test and the standard RPR 18-mm circle card tests for the serodiagnosis of syphilis. Sera from 638 individuals were used in this study. Two pilot lots and two production lots of antigen were submitted by Beckman Instruments, Inc., for evaluation. Qualitative agreement among the three RPR card tests was 98....

A survey on the occurrence of ochratoxin A in feeds and sera collected in conventional and organic poultry farms in Northern Italy

Directory of Open Access Journals (Sweden)

Laura Cavallarin

2010-01-01

Full Text Available A survey has been conducted on conventional and organic poultry farms located in northern Italy in order to investigate the occurrence of ochratoxin A (OTA in feeds and sera in 2006. Ten poultry farms were monitored by taking 20 samples of feed and 94 samples of blood. OTA was assessed through immunoaffinity column purification and HPLC analysis. For in-house validation, recovery experiments, carried out on the spiked samples in the range of 1.0-10.0 μg OTA kg-1 and 0.3-3.0 ng OTA ml-1 for the feed and serum samples, respectively, led to overall recovery averages of 80.6% (RDS=7.3%, n=9 and 83.3% (RDS=3.1%, n=9, respectively. All the feed samples were contaminated by OTA with values ranging from 0.04 to 6.50 μg kg-1. Fiftythree percent of the sera samples were positive, with values ranging from 0.003- 0.165 ng ml-1. None of the feed samples was above the limits set by the European Union on OTA contamination in poultry feeds. No statistically significant differences in OTA contamination of feed or sera were observed either between the organic vs conventional group or between the laying hens vs broiler group.

Solubilization of immune complexes in complement factor deficient sera and the influence of temperature, ionic strength and divalent cations on the solubilization reaction

DEFF Research Database (Denmark)

Baatrup, Gunnar; Petersen, Ivan; Svehag, Svend-Erik

1984-01-01

The complement-mediated solubilization (CMS) of immune complexes (IC) and the initial kinetics (IKS) of this reaction in human sera depleted of or deficient in C2, C3, C8, factors B, P and I were investigated. Sera depleted of B or P and those lacking native C3 or factor I showed virtually no CMS......M. Chelation of Ca2+ in serum by Mg2+-ethylene glycol tetraacetic acid reduced the CMS capacity by up to 50% and the IKS was markedly retarded. Varying the Zn2+ or Mn2+ ion concentrations in serum influenced neither the IKS nor the CMS capacity....

KAtex antigen-detection test as a diagnostic tool for latent visceral ...

African Journals Online (AJOL)

SERVER

2008-04-03

Apr 3, 2008 ... Urine samples were tested by kAtex kit to detect VL antigen. All sera .... IFA, and ELISA-IgG and IgM) to finding of agreement and validity and reliability .... Animal inoculation is not usually employed as a diag- nostic test, since ...

Solid-phase enzyme immunoassay or radioimmunoassay for the detection of immune complexes based on their recognition by conglutinin: conglutinin-binding test. A comparative study with /sup 125/I-labelled Clq binding and Raji-cell RIA tests

Energy Technology Data Exchange (ETDEWEB)

Casali, P; Bossus, A; Carpentier, N A; Lambert, P H [Hopital Cantonal Geneve (Switzerland)

1977-01-01

Bovine conglutinin was used in a solid-phase assay for the detection of immune complexes. In a first step, the tested serum sample was incubated in polypropylene tubes coated with conglutinin to allow C3-coated immune complexes to bind to solid-phase conglutinin. In a second step, the conglutinin-bound complexes were detected using an enzyme-conjugated or radiolabelled anti-immunoglobulin antibody. The conglutinin-binding (KgB) test did not suffer from the interference of DNA, heparin or endotoxins. Its limit of sensitivity for aggregated IgG was 3 ..mu..g/ml undiluted human serum. Immune complexes prepared in vitro using tetanus toxoid, or DNA, and corresponding antibodies in human sera could be detected at various antigen/antibody ratios and at antibody concentrations lower than 8 ..mu..g/ml. The KgB test allowed for the detection of immune complexes in sera from patients with systemic lupus erythematosus, rheumatoid arthritis, idiopathic vasculitis, leprosy and leukemia. These sera were also tested using the /sup 125/I-labelled Clq-binding activity (BA) test and the KgB test simultaneously, and a significant rank order correlation was observed. In patients with leukemia, a significant correlation was observed using three tests, KgB, /sup 125/I-labelled Clq BA and Raji-cell radioimmunoassay (RIA). Therefore, the KgB test appears as a simple and reproducible method, utilizing a very stable reagent, with a sensitivity and specificity comparable to the other tests studied and allowing for clinical application.

EPSTEIN-BARR VIRUS INFECTIONS – AVIDITY TEST FOR IgG ANTIBODIES

Directory of Open Access Journals (Sweden)

Katja Strašek

2001-06-01

Full Text Available Background. We wish to introduce specific IgG avidity test as a supplementary assay in serological screening for Epstein-Barr virus infection if the status of patient cannot be resolved from a single serum sample with routine testing.Methods. Avidity of IgG antibodies was determined in sera of 57 patients with different stage of Epstein-Barr virus infection. Enzyme-immuno assay was used with a short incubation of 6-molar urea included in the procedure. Urea should remove low avidity antibodies. Avidity was expressed as the avidity index. Avidity testing with commercial kit was done as well.Results. Low avidity index was found for IgG antibodies of acute phase sera and high for those of past infection, recent infection and reactivation of endogenic virus.Conclusions. Avidity test for IgG antibodies might be supplementary assay to prove acute infection but also to resolve some other clinical states related to Epstein-Barr virus.

Comparative efficacy of antigen and antibody detection tests for human trichinellosis

International Nuclear Information System (INIS)

Ivanoska, D.; Cuperlovic, K.; Gamble, H.R.; Murrell, K.D.

1989-01-01

Sera collected from patients with suspected or confirmed exposure to Trichinella spiralis were tested for circulating parasite antigens and antiparasite antibodies. Using an immunoradiometric assay, excretory--secretory antigens from muscle-stage larvae of T. spiralis were detected in the sera of 47% of 62 patients with clinical trichinellosis and 13% of 39 patients without clinical signs but suspected of exposure to infected meat. In comparison, antibodies were detected using an indirect immunofluorescent test in the circulation of 100% of the 62 patients with clinical trichinellosis and 46% of the 39 patients with suspected exposure. The presence of antibodies specific to excretory-secretory products of T. spiralis muscle larvae was confirmed in the majority of the samples tested by a monoclonal antibody-based competitive inhibition assay. These results indicate that antibody detection is a more sensitive diagnostic method for human trichinellosis, but that antigen detection might be a useful confirmatory test because it is a direct demonstration of parasite products in the circulation

Variant surface glycoproteins from Venezuelan trypanosome isolates are recognized by sera from animals infected with either Trypanosoma evansi or Trypanosoma vivax.

Science.gov (United States)

Camargo, Rocío; Izquier, Adriana; Uzcanga, Graciela L; Perrone, Trina; Acosta-Serrano, Alvaro; Carrasquel, Liomary; Arias, Laura P; Escalona, José L; Cardozo, Vanessa; Bubis, José

2015-01-15

Salivarian trypanosomes sequentially express only one variant surface glycoprotein (VSG) on their cell surface from a large repertoire of VSG genes. Seven cryopreserved animal trypanosome isolates known as TeAp-ElFrio01, TEVA1 (or TeAp-N/D1), TeGu-N/D1, TeAp-Mantecal01, TeGu-TerecayTrino, TeGu-Terecay03 and TeGu-Terecay323, which had been isolated from different hosts identified in several geographical areas of Venezuela were expanded using adult albino rats. Soluble forms of predominant VSGs expressed during the early infection stages were purified and corresponded to concanavalin A-binding proteins with molecular masses of 48-67 kDa by sodium dodecyl sulfate-polyacrylamide gel electropohoresis, and pI values between 6.1 and 7.5. The biochemical characterization of all purified soluble VSGs revealed that they were dimers in their native form and represented different gene products. Sequencing of some of these proteins yielded peptides homologous to VSGs from Trypanosoma (Trypanozoon) brucei and Trypanosoma (Trypanozoon) evansi and established that they most likely are mosaics generated by homologous recombination. Western blot analysis showed that all purified VSGs were cross-reacting antigens that were recognized by sera from animals infected with either T. evansi or Trypanosoma (Dutonella) vivax. The VSG glycosyl-phosphatidylinositol cross-reacting determinant epitope was only partially responsible for the cross-reactivity of the purified proteins, and antibodies appeared to recognize cross-reacting conformational epitopes from the various soluble VSGs. ELISA experiments were performed using infected bovine sera collected from cattle in a Venezuelan trypanosome-endemic area. In particular, soluble VSGs from two trypanosome isolates, TeGu-N/D1 and TeGu-TeracayTrino, were recognized by 93.38% and 73.55% of naturally T. vivax-infected bovine sera, respectively. However, approximately 70% of the sera samples did not recognize all seven purified proteins. Hence, the

A simple and economic slide micro-immunoenzymatic (Micro-SIA test for epidemiological studies of toxoplasmosis

Directory of Open Access Journals (Sweden)

Mara Rosenzvit

1994-03-01

Full Text Available A slide micro-immunoenzymatic assay (micro-SIA to detectantibodies to non-particulate Toxoplasma gondii antigens is described. This assay allows the diagnosis of toxoplasmosis infection in about 1 hr. Twenty-four determinations can be performed per slide. Five hundred ng of antigen and 5 or 10 µl drop of each reactive are necessary per well. The clear contrast of colours obtained for negative and positive sera after the test is finished, allows direct discrimination of the results. However, it is possible to quantify the results of the reaction using a minireader. Sera dilution cutoff value, determined as themost frequent titre for the general population, is 1:100. The toxoplasma micro-SIA correlates well with indirect immunofluorescence (IIF, its sensitivity is atleast three times as much as IIF. The test has an intra and inter assay variation coefficient of 5.46 per cent and of 6.24 per cent respectively. Sera obtained at random from argentinian people were analyzed and a 56 per cent of infection was found. The main features of the Toxoplasma micro-SIA are its simplicity, sensitivity, reproducibility, and the virtual absence of background making it very suitable for screening tests.

[Detection of anti-Leptospira antibodies in sera of patients in the latex agglutination test].

Science.gov (United States)

Volina, E G; Sarukhanova, L E; Iashina, N V; Prokopov, N I; Shkarlat, P E; Barysheva, I V

2001-01-01

The results of the preliminary evaluation of the sensitivity and specificity of the newly developed diagnostic test based on the determination of genus-specific antibodies to leptospires in the latex agglutination test, are presented. This test makes it possible to detect anti-Leptospira antibodies of any serogroup. The advantages of the developed test have been determined.

Factors affecting the serological testing of cadaveric donor cornea.

Science.gov (United States)

Raj, Anuradha; Mittal, Garima; Bahadur, Harsh

2018-01-01

The purpose of this study was to evaluate the serological profile of the eye donors and to study the influence of various factors on serological test results. A cross-sectional, observational study was conducted, and data of 509 donors were reviewed from the records of eye bank from December 2012 to June 2017. Various details of donors analyzed included the age, sex of the donor, cause of death, source of tissue, time since blood collection after death, macroscopic appearance of blood sample, and details of discarded tissues. Serological examination of blood was performed for human immunodeficiency virus (HIV), hepatitis B virus, hepatitis C virus (HCV), venereal disease research laboratory (VDRL), and serology reports reactive or nonreactive were analyzed. Among the 509 donors, 295 (58%) were male, and 420 (82.50%) belonged to age group ≥60 years. Most donors (354, 69.5%) died due to cardiac arrest. Macroscopically, sera were normal in the majority of 488 (95.9%) cases. Among 509 donors, 475 (93.3%) were nonreactive, 12 (2.4%) donors were found to be reactive to hepatitis B surface antigen (HBsAg), and 1 (0.2%) was reactive to HCV, but no donor serology was reactive to HIV or VDRL. Twenty-one (4.12%) donors' sera were not fit for serological testing. Among all donors, 475 (93.32%) donors were accepted and 34 (6.67%) were rejected or discarded on the basis of serological testing. Cause of death and macroscopic aspect of sera influenced the serological results in a highly significant manner (P = 0.00). Acceptance or rejection of the donor was significantly influenced by the serological results of the donor (P = 0.00). The seroprevalence among eye donor for HBsAg and HCV was 12 (2.4%) and 1 (0.2%), respectively. Factors such as cause of death and macroscopic aspect of sera influence the serological results. Time since blood collection or sampling will not show any impact on viral serological results if postmortem sampling will be done in donor cornea.

Pesquisa de aglutininas anti Brucella canis em soros humanos na cidade de São Paulo, Brasil Research on agglutinins for Brucella canis in human sera in the city of S. Paulo, Brazil

Directory of Open Access Journals (Sweden)

Maria Helena Matiko Akao Larsson

1980-09-01

Full Text Available De 330 soros humanos examinados pela prova de soroaglutinação lenta em tubos, 4(1,21% apresentaram aglutininas anti Brucella canis em diluição 1:100 (1 reagente com título 100, 2 reagentes com título 200 e 1 reagente com título 400.Of the 330 human sera tested by tube agglutination test, 4 (1.21% were positive for Brucella canis antibodies with tilers 1:100 or higher (1 reagent with titer of 1:100, 2 reagents with titer of 1:200, and 1 reagent with tiler of 1:400.

Study of the titers of Anti-Epstein-Barr virus antibodies in the sera of atomic bomb survivors

Energy Technology Data Exchange (ETDEWEB)

Akiyama, Mitoshi; Kusunoki, Yoichiro; Kyoizumi, Seishi; Ozaki, Kyoko; Mizuno, Shoichi; Cologne, J.B.

1993-12-31

Antibody titers to Epstein-Barr virus antigens were determined in the sera of 372 atomic bomb survivors to evaluate the effect of the previous radiation exposure on immune competence against the latent infection of the virus. The proportion of persons with high titers ({>=} 1:40) of IgG antibodies to the early antigen was significantly elevated in the exposed survivors. Furthermore, the distribution of IgM titers against the viral capsid antigen was significantly affected by radiation dose with an increased occurrence of titers of 1:5 and 1:10 in the exposed persons, although the dose effect was only marginally suggestive when persons with rheumatoid factor were eliminated from the analysis. These results suggest that reactivation of Epstein-Barr virus in the latent stage occurs more frequently in the survivors, even though this might not be affected by the radiation dose. Otherwise, there was neither an increased trend in the prevalence of high titers ({>=} 1:640) of IgG antibodies to the viral capsid antigen among the exposed people nor a correlation between the radiation exposure and distributions of titers of IgA antibodies to the viral capsid antigen or antibodies to the anti-Epstein-Barr virus-associated nuclear antigen. (author).

Study of the titers of Anti-Epstein-Barr virus antibodies in the sera of atomic bomb survivors

International Nuclear Information System (INIS)

Akiyama, Mitoshi; Kusunoki, Yoichiro; Kyoizumi, Seishi; Ozaki, Kyoko; Mizuno, Shoichi; Cologne, J.B.

1993-01-01

Antibody titers to Epstein-Barr virus antigens were determined in the sera of 372 atomic bomb survivors to evaluate the effect of the previous radiation exposure on immune competence against the latent infection of the virus. The proportion of persons with high titers (≥ 1:40) of IgG antibodies to the early antigen was significantly elevated in the exposed survivors. Furthermore, the distribution of IgM titers against the viral capsid antigen was significantly affected by radiation dose with an increased occurrence of titers of 1:5 and 1:10 in the exposed persons, although the dose effect was only marginally suggestive when persons with rheumatoid factor were eliminated from the analysis. These results suggest that reactivation of Epstein-Barr virus in the latent stage occurs more frequently in the survivors, even though this might not be affected by the radiation dose. Otherwise, there was neither an increased trend in the prevalence of high titers (≥ 1:640) of IgG antibodies to the viral capsid antigen among the exposed people nor a correlation between the radiation exposure and distributions of titers of IgA antibodies to the viral capsid antigen or antibodies to the anti-Epstein-Barr virus-associated nuclear antigen. (author)

Contribution to diagnostics/prognostics of tuberculosis in children. I. New methods of assaying zinc and simultaneously copper and zinc in diluted sera by flame atomic-absorption spectrometry

Directory of Open Access Journals (Sweden)

Luterotti Svjetlana

2015-09-01

Full Text Available In an attempt to provide a reliable status of metal ions in children, new methods of analysis of children’s sera are proposed. New flame atomic-absorption spectrometric (FAAS methods are simple, cost- and time-effective and, above all, labor-, reagent- and sample-saving. Two methods were suggested: method A for simultaneous determination of Cu and Zn from 5-fold diluted sera, and method B, for assaying zinc alone in 10-fold diluted samples. Both methods are based on a single-step sample pretreatment (deproteinization with 3 mol dm–3 HCl. Method A uses a single-step calibration with a mixed standard. The main advantage of method B is an additional reduction in sample consumption. Both methods were fully validated against reference methods. Accuracy, sensitivity and precision have proven them to be comparable to the reference methods in terms of analytical performance, and applicable to analyses of children’s sera.

Neutron activation analysis of Cl, K and Na content in whole blood of horses used in hyperimmune sera production

International Nuclear Information System (INIS)

Baptista, T.S.; Zamboni, C.B.; Medeiros, J.A.G.; Freitas, M.G.; Higashi, H.G.; Marcelino, J.R.

2009-01-01

Using neutron activation analysis technique Cl, K and Na concentration were obtained in whole blood of equines used for antivenom production at Butantan Institute (Sao Paulo, Brazil). These data were compared with the human whole blood estimation. No significant difference was observed suggesting that this model animal is adequate sera production. (author)

İlköğretim Öğretmen Adaylarının Sera Etkisi İle İlgili Kavram Yanılgıları

OpenAIRE

Arsal, Zeki

2010-01-01

Bu çalışmanın amacı ilköğretim fen bilgisi ve sınıf öğretmenliği kavram yanılgılarını döndürmek. Araştırma eğitim fakültesi ilköğretim fen bilgisi ve sınıf öğretmenliği bölümler öğrenim gören 171 öğretmen adayı ile yapılmış. Çalışmada sera etkisinin nedenleri, sonuçları ve sera açısından önleme yollarıyla ilgili kavram yanılgıları araştırılmıştır. Araştırmanın anlamı toplanması "Sera Etkisi le lgili Düşünceler Anketi" Anket üç alt boyuttan ve yirmi üç maddeden. Deneme hakkı hem fen ...

Differences in immunoglobulin subclasses between dye exclusion and 51Cr release complement-dependent lymphocytotoxicity assays

International Nuclear Information System (INIS)

McConnachie, P.R.; Denegri, J.F.; Lower, F.E.; Torry, D.S.; McIntyre, J.A.

1986-01-01

Paradoxical differences previously noted between lymphocytotoxicity detected by dye exclusion at room temperature (CDCE) or by 51 Cr release (CDC 51 Cr) at 37 0 C in maternal antipaternal complement-dependent lymphocytotoxicity have suggested that CDCE and CDC 51 Cr at 37 0 C, but not at 20 degrees C, may detect different immunological antibody-antigen interactions. Reactions in the two test systems against the same target cells were compared in sera from known immune dialysis patients, secondary aborting women, and refractory platelet recipients before and after heat treatment of sera, absorption with solid-phase heparin, anti-light-chain augmentation, and the addition of murine monoclonal anti-IgG subclass antibodies. The results demonstrate significant differences between the two tests using the same target and sera. Further, the results imply the presence of an inhibitor and an inhibitor of inhibitor in sera. The involvement of different immunoglobulin subclasses was shown in the two tests. These data demonstrate the necessity for further study of the nature of the differences in the mechanisms of these clinically important antibody-detecting systems

An ELISA-inhibition test using monoclonal antibody for the serology of leprosy

NARCIS (Netherlands)

Klatser, P. R.; de Wit, M. Y.; Kolk, A. H.

1985-01-01

In this study a mouse monoclonal antibody (47-9) is described, which recognized an epitope on the 36 kD protein antigen of M. leprae. The monoclonal antibody showed specificity for M. leprae. An ELISA-inhibition test based on the competitive inhibition by antibodies from human test sera of the

Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern.

Science.gov (United States)

Aisa, M J; Castillejo, S; Gallego, M; Fisa, R; Riera, M C; de Colmenares, M; Torras, S; Roura, X; Sentis, J; Portus, M

1998-02-01

Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.

Evaluating enzootic bovine leukemia virus infection by means of molecular probe compared with the results of serological tests

International Nuclear Information System (INIS)

Reichert, M.; Grundbock, J.; Rulka, J.; Kozaczynska, B.; Stec, J.

1994-01-01

The present studies were aimed at determining the relation between the finding obtained by means of serological tests and the specific molecular probe. Serological tests were performed according to the methods recommended by the Polish Ministry of Agriculture; ELISA was run with ''Bioveta'' and ''Rhone Merieux'' kits and the AGID test was performed with EBL antigen made in our laboratory. The molecular probe was prepared from the previously cloned provirus DNA of EBL virus. The EBL provirus was detected in 28 samples taken from 44 randomly selected cows in three herds on which a leukemia eradication programme was in process. Three sera out of 28 positive reacting animals were negative in AGID test and only one serum in ELISA. The results indicate that the use of a specific molecular probe has some advantages in the diagnosis of latent virus infections. Besides, it can be applied in the studies on the pathogenesis of enzootic bovine leukemia. (author). 13 refs, 1 fig., 1 tab

Correlates of previous couples’ HIV counseling and testing uptake among married individuals in three HIV prevalence strata in Rakai, Uganda

Directory of Open Access Journals (Sweden)

Joseph K. B. Matovu

2015-06-01

Full Text Available Background: Studies show that uptake of couples’ HIV counseling and testing (couples’ HCT can be affected by individual, relationship, and socioeconomic factors. However, while couples’ HCT uptake can also be affected by background HIV prevalence and awareness of the existence of couples’ HCT services, this is yet to be documented. We explored the correlates of previous couples’ HCT uptake among married individuals in a rural Ugandan district with differing HIV prevalence levels. Design: This was a cross-sectional study conducted among 2,135 married individuals resident in the three HIV prevalence strata (low HIV prevalence: 9.7–11.2%; middle HIV prevalence: 11.4–16.4%; and high HIV prevalence: 20.5–43% in Rakai district, southwestern Uganda, between November 2013 and February 2014. Data were collected on sociodemographic and behavioral characteristics, including previous receipt of couples’ HCT. HIV testing data were obtained from the Rakai Community Cohort Study. We conducted multivariable logistic regression analysis to identify correlates that are independently associated with previous receipt of couples’ HCT. Data analysis was conducted using STATA (statistical software, version 11.2. Results: Of the 2,135 married individuals enrolled, the majority (n=1,783, 83.5% had been married for five or more years while (n=1,460, 66% were in the first-order of marriage. Ever receipt of HCT was almost universal (n=2,020, 95%; of those ever tested, (n=846, 41.9% reported that they had ever received couples’ HCT. There was no significant difference in previous receipt of couples’ HCT between low (n=309, 43.9%, middle (n=295, 41.7%, and high (n=242, 39.7% HIV prevalence settings (p=0.61. Marital order was not significantly associated with previous receipt of couples’ HCT. However, marital duration [five or more years vis-à-vis 1–2 years: adjusted odds ratio (aOR: 1.06; 95% confidence interval (95% CI: 1.04–1.08] and

Evaluation of the LDBIO point of care test for the combined detection of toxoplasmic IgG and IgM.

Science.gov (United States)

Chapey, Emmanuelle; Wallon, Martine; Peyron, François

2017-01-01

The toxoplasma ICT IgG-IgM rapid diagnostic test for the simultaneous detection of specific toxoplasmic immunoglobulin (Ig) G and IgM was compared with the Architect fully automated chemiluminescence test. Four hundred sera were included, among which 248 scored negative in Architect. The cassettes were easily read with the naked eye. Diagnostic sensitivity and specificity were 97% and 96%, respectively. The test scored 8 false-positive IgG and yielded negative results in 3 sera displaying unspecific IgM in Architect. The LDBIO appears to be a reliable first line test, although the false-positive results for IgG deserve further investigation. Such an easily performed test could be used advantageously for screening for toxoplasmosis in pregnant women. Copyright © 2016 Elsevier B.V. All rights reserved.

Autoantibodies to IL-1 alpha in sera from umbilical cords, children, and adults, and from patients with juvenile chronic arthritis

DEFF Research Database (Denmark)

Müller, K; Hansen, M B; Zak, M

1996-01-01

umbilical cords (n = 11), children (n = 45), and adults (n = 20), as well as in 51 patients with juvenile chronic arthritis (JCA) of pauciarticular (n = 34), polyarticular (n = 8), or systemic onset type (n = 9). RESULTS. The frequency of positive sera was significantly lower in children than in cord blood...

Levels of inhibitors of tumor necrosis factor alpha and interleukin 1beta in urine and sera of patients with urosepsis

NARCIS (Netherlands)

Olszyna, D. P.; Prins, J. M.; Buis, B.; van Deventer, S. J.; Speelman, P.; van der Poll, T.

1998-01-01

The antiinflammatory cytokine response during urosepsis was determined by measurement of concentrations of soluble tumor necrosis factor receptor (sTNFR) types I and II, interleukin 1 receptor antagonist (IL-1ra), soluble IL-1 receptor type II (sIL-1RII), and interleukin 10 in sera and urine of 30

Evaluation of a bovine antibody test for diagnosing Mycobacterium avium complex in patients with cystic fibrosis

DEFF Research Database (Denmark)

Qvist, Tavs; Pressler, Tacjana; Katzenstein, Terese L.

2017-01-01

Introduction: The aim of this study was to test a commercial bovine enzyme-linked immunosorbent assay for investigating antibody activity against Mycobacterium avium complex. Methods: All patients at the Copenhagen Cystic Fibrosis (CF) Center who had culture for nontuberculous mycobacteria...... before and after culture conversion was performed in case patients. Results: Out of 286 included subjects, six had clinical M. avium complex pulmonary disease at the time of sera sampling. These patients presented with higher antibody test values (P-value ... at ruling out pulmonary disease. Screening sera from patients with CF could guide clinicians to focus attention on patients at higher risk of M. avium complex pulmonary disease. Pediatr Pulmonol. 2017;52:34–40....

Comparative 1H NMR-based metabonomic analysis of HIV-1 sera

International Nuclear Information System (INIS)

Philippeos, C.; Steffens, F. E.; Meyer, D.

2009-01-01

1 H NMR spectroscopy of sera from HIV-1 infected and uninfected individuals was performed on 300 and 600 MHz instruments. The resultant spectra were automatically data reduced to 90 and 180 integral segments of equal length. Analysis of variance identified significant differences between the sample groups, especially for the samples analyzed on 600 MHz and reduced to fewer segments. Linear discriminant analysis correctly classified 100% of the samples analyzed on the 300 MHz NMR (reduced to 180 segments); an increase in instrument sensitivity resulted in lower percentages of correctly classified samples. Multinomial logistic regression (MLR) resulted in 100% correct classification of all samples from both instruments. Thus 1 H-NMR metabonomics on either instrument distinguishes HIV-positive individuals using or not using anti retroviral therapy, but the sensitivity of the instrument impacts on data reduction. Furthermore, MLR is a novel multivariate statistical technique for improved classification of biological data analyzed in NMR

Proteomic data show an increase in autoantibodies and alpha-fetoprotein and a decrease in apolipoprotein A-II with time in sera from senescence-accelerated mice

Energy Technology Data Exchange (ETDEWEB)

Guo, S.J. [Beijing Institute of Pharmacology and Toxicology, Beijing (China); Shanghai Key Laboratory of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Qi, C.H.; Zhou, W.X.; Zhang, Y.X. [Beijing Institute of Pharmacology and Toxicology, Beijing (China); Zhang, X.M.; Wang, J.; Wang, H.X. [National Center of Biomedical Analysis, Beijing (China)

2013-04-12

We evaluated changes in levels by comparing serum proteins in senescence-accelerated mouse-prone 8 (SAMP8) mice at 2, 6, 12, and 15 months of age (SAMP8-2 m, -6 m, -12 m, -15 m) to age-matched SAM-resistant 1 (SAMR1) mice. Mice were sacrificed, and blood was analyzed by 2-dimensional electrophoresis combined with mass spectrometry. Five protein spots were present in all SAMP8 serum samples, but only appeared in SAMR1 samples at 15 months of age except for spot 3, which also showed a slight expression in SAMR1-12 m sera. Two proteins decreased in the sera from SAMP8-2 m, -6 m, and -12 m mice, and divided into 2 spots each in SAMP8-15 m sera. Thus, the total number of altered spots in SAMP8 sera was 7; of these, 4 were identified as Ig kappa chain V region (M-T413), chain A of an activity suppressing Fab fragment to cytochrome P450 aromatase (32C2-A), alpha-fetoprotein, and apolipoprotein A-II. M-T413 is a monoclonal CD4 antibody, which inhibits T cell proliferation. We found that M-T413 RNA level was significantly enhanced in splenocytes from SAMP8-2 m mice. This agreed with serum M-T413 protein alterations and a strikingly lower blood CD4{sup +} T cell count in SAMP8 mice when compared to the age-matched SAMR1 mice, with the latter negatively correlating with serum M-T413 protein volume. Age-related changes in serum proteins favored an increase in autoantibodies and alpha-fetoprotein and a decrease of apolipoprotein A-II, which occurred in SAMP8 mice at 2 months of age and onwards. These proteins may serve as candidate biomarkers for early aging.

Proteomic data show an increase in autoantibodies and alpha-fetoprotein and a decrease in apolipoprotein A-II with time in sera from senescence-accelerated mice

International Nuclear Information System (INIS)

Guo, S.J.; Qi, C.H.; Zhou, W.X.; Zhang, Y.X.; Zhang, X.M.; Wang, J.; Wang, H.X.

2013-01-01

We evaluated changes in levels by comparing serum proteins in senescence-accelerated mouse-prone 8 (SAMP8) mice at 2, 6, 12, and 15 months of age (SAMP8-2 m, -6 m, -12 m, -15 m) to age-matched SAM-resistant 1 (SAMR1) mice. Mice were sacrificed, and blood was analyzed by 2-dimensional electrophoresis combined with mass spectrometry. Five protein spots were present in all SAMP8 serum samples, but only appeared in SAMR1 samples at 15 months of age except for spot 3, which also showed a slight expression in SAMR1-12 m sera. Two proteins decreased in the sera from SAMP8-2 m, -6 m, and -12 m mice, and divided into 2 spots each in SAMP8-15 m sera. Thus, the total number of altered spots in SAMP8 sera was 7; of these, 4 were identified as Ig kappa chain V region (M-T413), chain A of an activity suppressing Fab fragment to cytochrome P450 aromatase (32C2-A), alpha-fetoprotein, and apolipoprotein A-II. M-T413 is a monoclonal CD4 antibody, which inhibits T cell proliferation. We found that M-T413 RNA level was significantly enhanced in splenocytes from SAMP8-2 m mice. This agreed with serum M-T413 protein alterations and a strikingly lower blood CD4 + T cell count in SAMP8 mice when compared to the age-matched SAMR1 mice, with the latter negatively correlating with serum M-T413 protein volume. Age-related changes in serum proteins favored an increase in autoantibodies and alpha-fetoprotein and a decrease of apolipoprotein A-II, which occurred in SAMP8 mice at 2 months of age and onwards. These proteins may serve as candidate biomarkers for early aging

Factors affecting the serological testing of cadaveric donor cornea

Directory of Open Access Journals (Sweden)

Anuradha Raj

2018-01-01

Full Text Available Purpose: The purpose of this study was to evaluate the serological profile of the eye donors and to study the influence of various factors on serological test results. Methods: A cross-sectional, observational study was conducted, and data of 509 donors were reviewed from the records of eye bank from December 2012 to June 2017. Various details of donors analyzed included the age, sex of the donor, cause of death, source of tissue, time since blood collection after death, macroscopic appearance of blood sample, and details of discarded tissues. Serological examination of blood was performed for human immunodeficiency virus (HIV, hepatitis B virus, hepatitis C virus (HCV, venereal disease research laboratory (VDRL, and serology reports reactive or nonreactive were analyzed. Results: Among the 509 donors, 295 (58% were male, and 420 (82.50% belonged to age group ≥60 years. Most donors (354, 69.5% died due to cardiac arrest. Macroscopically, sera were normal in the majority of 488 (95.9% cases. Among 509 donors, 475 (93.3% were nonreactive, 12 (2.4% donors were found to be reactive to hepatitis B surface antigen (HBsAg, and 1 (0.2% was reactive to HCV, but no donor serology was reactive to HIV or VDRL. Twenty-one (4.12% donors' sera were not fit for serological testing. Among all donors, 475 (93.32% donors were accepted and 34 (6.67% were rejected or discarded on the basis of serological testing. Cause of death and macroscopic aspect of sera influenced the serological results in a highly significant manner (P = 0.00. Acceptance or rejection of the donor was significantly influenced by the serological results of the donor (P = 0.00. Conclusion: The seroprevalence among eye donor for HBsAg and HCV was 12 (2.4% and 1 (0.2%, respectively. Factors such as cause of death and macroscopic aspect of sera influence the serological results. Time since blood collection or sampling will not show any impact on viral serological results if postmortem sampling

Sensitive and specific detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)—Specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in μ-capture and IgG immune complex (IC) ELISA tests

Science.gov (United States)

Emmerich, Petra; Mika, Angela; von Possel, Ronald; Rackow, Anne; Liu, Yang; Schmitz, Herbert; Sherifi, Kurtesh; Halili, Barie; Jakupi, Xhevat; Berisha, Lindita; Ahmeti, Salih

2018-01-01

As the most widespread tick-borne arbovirus causing infections in numerous countries in Asia, Africa and Europe, Crimean-Congo Hemorrhagic Fever Virus (CCHFV, family Nairoviridae) was included in the WHO priority list of emerging pathogens needing urgent Research & Development attention. To ensure preparedness for potential future outbreak scenarios, reliable diagnostic tools for identification of acute cases as well as for performance of seroprevalence studies are necessary. Here, the CCHFV ortholog of the major bunyavirus antigen, the nucleoprotein (NP), was recombinantly expressed in E.coli, purified and directly labeled with horseradish peroxidase (HRP). Employing this antigen, two serological tests, a μ-capture ELISA for the detection of CCHFV-specific IgM antibodies (BLACKBOX CCHFV IgM) and an IgG immune complex (IC) ELISA for the detection of CCHFV-specific IgG antibodies (BLACKBOX CCHFV IgG), were developed. Test performance was evaluated and compared with both in-house gold standard testing by IgM/IgG indirect immunofluorescence (IIF) and commercially available ELISA tests (VectoCrimean-CHF-IgM/IgG, Vector-Best, Russia) using a serum panel comprising paired samples collected in Kosovo during the years 2013–2016 from 15 patients with an acute, RT-PCR-confirmed CCHFV infection, and 12 follow-up sera of the same patients collected approximately one year after having overcome the infection. Reliably detecting IgM antibodies in all acute phase sera collected later than day 4 after onset of symptoms, both IgM ELISAs displayed excellent diagnostic and analytical sensitivity (100%, 95% confidence interval (CI): 85.2%–100.0%). While both IgG ELISAs readily detected the high IgG titers present in convalescent patients approximately one year after having overcome the infection (sensitivity 100%, 95% CI: 73.5%–100.0%), the newly developed BLACKBOX CCHFV IgG ELISA was superior to the commercial IgG ELISA in detecting the rising IgG titers during the acute phase

Application of indirect immunofluorescent test with an improved HEp-2 substrate tranfected with human Ro60/SSA autoantigens

International Nuclear Information System (INIS)

Lv Liangjing; Chen Shunle; Gu Yueying; Shen Nan; Bao Chunde; Wang Yuan; Xue Feng; Ye Peng; Yu Chongzhao

2006-01-01

To develop an improved substrate for indirect immunofluorescent test (IIF) to detect anti-Ro/SSA autoantibodies, the human 60-kDa Ro/SSA autoantigens (Ro60) cDNAs were obtained from placental cDNA library using PCR and were cloned into the mammalian expression vectorpEGFP-C1. Then the recombinant plasmids were transfected into HEp-2 cells. We con- firmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of fluorescence microscopy, immunoblotting and IIF. HEp-2 and HEp-Ro60 was analyzed by IIF using a panel of 10 precipitinpositive anti-Ro human sera simultaneously. Stable expression of Ro60-GFP (green fluorescent protein) fusion proteins maintained ten more generations. And Ro60-GFP kept the antigenicity of Ro and had its own characteristic immunofluorescent pattern in HEp-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer, P<0.01). Eight (8/10) positive an- ti-Ro sera showed characteristic immunofluorescent pattern on HEp-Ro60, including two sera which were antinuclear antibodies (ANA) negative on untransfected HEp-2. IIF-ANA in all healthy sera were negative on HEp-Ro60. As a kind of new substrate of IIF, the Ro60 transfectants can be used to detect anti-Ro antibodies. In addition, transfected HEp-2 cells kept the immunofluorescent property of HEp-2 cells in IIF-ANA tests and could be employed as substrate for the routine IIF-ANA detection. The method improved the sensitivity of IIF-ANA. (authors)

Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips

Science.gov (United States)

Jara, Rocio F.; Sepúlveda, Carolina; Ip, Hon S.; Samuel, Michael D.

2015-01-01

Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

Solid-phase enzyme immunoassay or radioimmunoassay for the detection of immune complexes based on their recognition by conglutinin: conglutinin-binding test

International Nuclear Information System (INIS)

Casali, P.; Bossus, A.; Carpentier, N.A.; Lambert, P.H.

1977-01-01

Bovine conglutinin was used in a solid-phase assay for the detection of immune complexes. In a first step, the tested serum sample was incubated in polypropylene tubes coated with conglutinin to allow C3-coated immune complexes to bind to solid-phase conglutinin. In a second step, the conglutinin-bound complexes were detected using an enzyme-conjugated or radiolabelled anti-immunoglobulin antibody. The conglutinin-binding (KgB) test did not suffer from the interference of DNA, heparin or endotoxins. Its limit of sensitivity for aggregated IgG was 3 μg/ml undiluted human serum. Immune complexes prepared in vitro using tetanus toxoid, or DNA, and corresponding antibodies in human sera could be detected at various antigen/antibody ratios and at antibody concentrations lower than 8 μg/ml. The KgB test allowed for the detection of immune complexes in sera from patients with systemic lupus erythematosus, rheumatoid arthritis, idiopathic vasculitis, leprosy and leukemia. These sera were also tested using the 125 I-labelled Clq-binding activity (BA) test and the KgB test simultaneously, and a significant rank order correlation was observed. In patients with leukemia, a significant correlation was observed using three tests, KgB, 125 I-labelled Clq BA and Raji-cell radioimmunoassay (RIA). Therefore, the KgB test appears as a simple and reproducible method, utilizing a very stable reagent, with a sensitivity and specificity comparable to the other tests studied and allowing for clinical application. (author)

SERA Scenarios of Early Market Fuel Cell Electric Vehicle Introductions: Modeling Framework, Regional Markets, and Station Clustering; NREL (National Renewable Energy Laboratory)

Energy Technology Data Exchange (ETDEWEB)

Melaina, M.

2015-03-23

This presentation provides an overview of the Scenario Evaluation and Regionalization Analysis (SERA) model, describes the methodology for developing scenarios for hydrogen infrastructure development, outlines an example "Hydrogen Success" scenario, and discusses detailed scenario metrics for a particular case study region, the Northeast Corridor.

9 CFR 147.7 - Standard test procedures for mycoplasma. 5

Science.gov (United States)

2010-01-01

... depend on preference, laboratory facilities, and availability of antigen. These three tests, though quite... titered sera are available from the National Veterinary Services Laboratories (NVSL), P.O. Box 884, Ames... blood to 5 parts Alsever's solution (e.g., 8 ml blood in 40 ml of Alsever's solution). Centrifuge the...

A novel synthetic peptide microarray assay detects Chlamydia species-specific antibodies in animal and human sera.

Science.gov (United States)

Sachse, Konrad; Rahman, Kh Shamsur; Schnee, Christiane; Müller, Elke; Peisker, Madlen; Schumacher, Thomas; Schubert, Evelyn; Ruettger, Anke; Kaltenboeck, Bernhard; Ehricht, Ralf

2018-03-16

Serological analysis of Chlamydia (C.) spp. infections is still mainly based on micro-immunofluorescence and ELISA. To overcome the limitations of conventional serology, we have designed a novel microarray carrying 52 synthetic peptides representing B-cell epitopes from immunodominant proteins of all 11 chlamydial species. The new assay has been validated using monospecific mouse hyperimmune sera. Subsequently, serum samples from cattle, sheep and humans with a known history of chlamydial infection were examined. For instance, the specific humoral response of sheep to treatment with a C. abortus vaccine has been visualized against a background of C. pecorum carriership. In samples from humans, dual infection with C. trachomatis and C. pneumoniae could be demonstrated. The experiments revealed that the peptide microarray assay was capable of simultaneously identifying specific antibodies to each Chlamydia spp. The actual assay represents an open platform test that can be complemented through future advances in Chlamydia proteome research. The concept of the highly parallel multi-antigen microarray proven in this study has the potential to enhance our understanding of antibody responses by defining not only a single quantitative response, but also the pattern of this response. The added value of using peptide antigens will consist in unprecedented serodiagnostic specificity.

Caracterização bioquímica de soros de bezerros utilizados na manutenção de culturas celulares usadas em virologia Biochemical characterization of calf sera used for the growth of cell cultures used in virology

Directory of Open Access Journals (Sweden)

Elisabeth Christina Nunes Tenório

1989-04-01

Full Text Available Oito lotes de soros de bezerros com idade maior ou menor de seis meses, usados para suplementar meios de cultivo de linhagens de células de uso difundido em virologia, foram testados quanto à sua capacidade promotora de crescimento (CPC e classificados como bons promotores ou promotores pobres. Foram pesquisados parâmetros relacionados com macronutrientes para estabelecer o perfil bioquímico daqueles lotes. Os resultados obtidos podem ser considerados valores normais para animais aparentemente sadios. As variações que ocorreram para um mesmo parâmetro bioquímico, entre os resultados da pesquisa e os de alguns estudos existentes na literatura, podem ser atribuídas à metodologia utilizada, à raça e idade dos animais, ou mesmo à própria dieta regional. A análise estatística, realizada pela aplicação do teste "t" de Student aos valores das médias e dos desvios padrão, demonstrou que, com relação às frações séricas, não ocorreram diferenças significantes entre soros de CPC celular boa ou pobre, considerando-se t c=2,45, enquanto que, para soros animais maiores ou menores de seis meses, apenas os resultados relativos às frações alfa e beta (t=2,68 e 2,61, respectivamente apresentaram significância, sendo superiores ao t crítico. Ficou evidenciado que a avaliação do conjunto de parâmetros bioquímicos pesquisados não permite diferenciar soros pertencentes aos dois grupos de animais, isto é, identificar soros de boa ou de pobre CPC, ou soros de bezerros maiores ou menores de seis meses de idadeEight lots of the calf sera employed to supplement culture media for the cultivation of animal cells, of widespread use in virology obtained from calves above and below six months of age were rated as good or as poor cell growth promoters according to their growth promoting capacity (GPC. Parameters related to macronutrients contained in these serum lots were then evaluated with the purpose of establishing their biochemical

Immunological reactive rate to Zika virus in canine sera: A report from a tropical area and concern on pet, zoonosis and reservoir host

Institute of Scientific and Technical Information of China (English)

Beuy Joob; Viroj Wiwanitkit

2017-01-01

This letter to editor discusses and describes Zika virus IgG seropositivity rate in canine sera from endemic area. This is the first world report and it raises an urgent concern on pet, zoonosis and reservoir host regarding Zika virus.

Surveillance of bluetongue virus antibody in goats using a recombinant VP7-based indirect ELISA in the coastal saline area of West Bengal, India

Directory of Open Access Journals (Sweden)

Raj K. Singh

2009-06-01

Full Text Available The authors describe the serological surveillance of bluetongue virus (BTV group-specific antibody in goats of the coastal saline (Sunderban area of West Bengal, India. A recombinant viral protein 7 (rVP7-based indirect enzyme-linked immunosorbent assay (ELISA was used to detect the antibody in sera. The bacterially expressed rVP7 was purified by affinity chromatography. The diagnostic performance of the assay was assessed by comparing it to the commercially available previously validated competitive ELISA. Using the control and 1 202 test sera, the cut-off value, sensitivity and specificity as well as other performance characteristics e.g. the Youden index, efficiency, positive and negative predictive value and prevalence were estimated. Field-collected goat sera (n = 1 202 were tested and a serological prevalence rate of 47% was observed in the study area.

Diagnosis of mold allergy by RAST and skin prick testing.

Science.gov (United States)

Nordvall, S L; Agrell, B; Malling, H J; Dreborg, S

1990-11-01

Sera from 33 patients with mold allergy proven by bronchial provocation were analyzed for specific IgE against six mold species comparing an improved Phadebas RAST with four other techniques. The new method was more sensitive and gave significantly higher IgE antibody concentrations for all tested molds except Cladosporium herbarum.

Comparison of agal gel precipitation test (AGPT) and enzyme linked ...

African Journals Online (AJOL)

The use of agar gel precipitation test (AGPT) and enzyme linked immunosorbent assay (ELISA) in assaying for the presence of infectious bursal disease (IBD) virus antibody in village chickens in Oyo State, Nigeria, was compared. Out of 400 sera subjected to ELISA, 360 (90%) samples where positive for IBD virus ...

Determination of IgE antibodies to the benzylpenicilloyl determinant: a comparison of the sensitivity and specificity of three radio allergo sorbent test methods.

Science.gov (United States)

Garcia, J J; Blanca, M; Moreno, F; Vega, J M; Mayorga, C; Fernandez, J; Juarez, C; Romano, A; de Ramon, E

1997-01-01

The quantitation of in vitro IgE antibodies to the benzylpenicilloyl determinant (BPO) is a useful tool for evaluating suspected penicillin allergic subjects. Although many different methods have been employed, few studies have compared their diagnostic specificity and sensitivity. In this study, the sensitivity and specificity of three different radio allergo sorbent test (RAST) methods for quantitating specific IgE antibodies to the BPO determinant were compared. Thirty positive control sera (serum samples from penicillin allergic subjects with a positive clinical history and a positive penicillin skin test) and 30 negative control sera (sera from subjects with no history of penicillin allergy and negative skin tests) were tested for BPO-specific IgE antibodies by RAST using three different conjugates coupled to the solid phase: benzylpenicillin conjugated to polylysine (BPO-PLL), benzylpenicillin conjugated to human serum albumin (BPO-HSA), and benzylpenicillin conjugated to an aminospacer (BPO-SP). Receiver operator control curves (ROC analysis) were carried out by determining different cut-off points between positive and negative values. Contingence tables were constructed and sensitivity, specificity, negative predictive values (PV-), and positive predictive values (PV+) were calculated. Pearson correlation coefficients (r) and intraclass correlation coefficients (ICC) were determined and the differences between methods were compared by chi 2 analysis. Analysis of the areas defined by the ROC curves showed statistical differences among the three methods. When cut-off points for optimal sensitivity and specificity were chosen, the BPO-HSA assay was less sensitive and less specific and had a lower PV- and PV+ than the BPO-PLL and BPO-SP assays. Assessment of r and ICC indicated that the correlation was very high, but the concordance between the PLL and SP methods was higher than between the PLL and HSA or SP and HSA methods. We conclude that for quantitating Ig

Development and evaluation of a competitive ELISA using a monoclonal antibody for antibody detection after goose parvovirus virus-like particles (VLPs) and vaccine immunization in goose sera.

Science.gov (United States)

Wang, Qian; Ju, Huanyu; Li, Yanwei; Jing, Zhiqiang; Guo, Lu; Zhao, Yu; Ma, Bo; Gao, Mingchun; Zhang, Wenlong; Wang, Junwei

2014-12-01

An assay protocol based on a monoclonal antibody-based competitive enzyme-linked immunosorbent assay (MAb-based C-ELISA) for detecting antibodies against goose parvovirus (GPV) and its virus-like particles (VLPs) is described. The assay was developed using baculovirus-expressed recombinant VP2 virus-like particles (rVP2-VLPs) as antigens and a monoclonal antibody against GPV as the competitive antibody. Of the four anti-GPV MAbs that were screened, MAb 1G3 was selected as it was blocked by the GPV positive serum. Based on the distribution of percent inhibition (PI) of the known negative sera (n=225), a cut-off value was set at 36% inhibition. Using this cut-off value, the sensitivity of the assay was 93.3% and the specificity was 95.8%, as compared with the gold standard (virus neutralization assay). The rVP2-VLPs did not react with anti-sera to other goose pathogens, indicating that it is specific for the recognization of goose parvovirus antibodies. The assay was then validated with serum samples from goslings vaccinated with several VLPs (rVP1-VLPs, rVP2-VLPs, rVP3-VLPs, and rCGV-VLPs) and other vaccines (inactivated and attenuated). The C-ELISA described in this study is a sensitive and specific diagnostic test and should have wide applications for the sero-diagnosis and immunologic surveillance of GPV. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of two solid-phase radioimmunoassay systems and a reverse passive haemagglutination test for the detection of hepatitis B surface antigen

International Nuclear Information System (INIS)

Hui, Z.; Coulepis, A.G.; Gust, I.D.

1982-01-01

The sensitivity and specificity of two commercially available radioimmunosay tests (Austria II-125, Abbott Laboratories; and International CIS, Commissariat Alenergie Atomique-Oris Laboratoire des Produits Biomedicaux) and a reverse passive haemagglutination test (Hepatest, Wellcome) for detection of hepatitis B surface antigen were evaluated using the Australian hepatitis B reference panel of 25 sera, and a panel of 257 sera collected from patients with acute hepatitis B, chronic carriers of hepatitis B surface antigen and two populations in which hepatitis B virus infection is known to be endemic. The three techniques were found to be generally comparable in sensitivity and specificity. The advantages and disadvantages of each method are discussed

Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

International Nuclear Information System (INIS)

Pastrana, Diana V.; Buck, Christopher B.; Pang, Y.-Y. S.; Thompson, Cynthia D.; Castle, Philip E.; FitzGerald, Peter C.; Krueger Kjaer, Susanne; Lowy, Douglas R.; Schiller, John T.

2004-01-01

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies

Comparative {sup 1}H NMR-based metabonomic analysis of HIV-1 sera

Energy Technology Data Exchange (ETDEWEB)

Philippeos, C. [University of Johannesburg, Department of Biochemistry (South Africa); Steffens, F. E. [University of Pretoria, Department of Statistics (South Africa); Meyer, D. [University of Pretoria, Department of Biochemistry (South Africa)], E-mail: debra.meyer@up.ac.za

2009-07-15

{sup 1}H NMR spectroscopy of sera from HIV-1 infected and uninfected individuals was performed on 300 and 600 MHz instruments. The resultant spectra were automatically data reduced to 90 and 180 integral segments of equal length. Analysis of variance identified significant differences between the sample groups, especially for the samples analyzed on 600 MHz and reduced to fewer segments. Linear discriminant analysis correctly classified 100% of the samples analyzed on the 300 MHz NMR (reduced to 180 segments); an increase in instrument sensitivity resulted in lower percentages of correctly classified samples. Multinomial logistic regression (MLR) resulted in 100% correct classification of all samples from both instruments. Thus {sup 1}H-NMR metabonomics on either instrument distinguishes HIV-positive individuals using or not using anti retroviral therapy, but the sensitivity of the instrument impacts on data reduction. Furthermore, MLR is a novel multivariate statistical technique for improved classification of biological data analyzed in NMR.

The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests.

Science.gov (United States)

Emmerzaal, A; de Wit, J J; Dijkstra, Th; Bakker, D; van Zijderveld, F G

2002-02-01

The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT.

A novel assay for discovery and characterization of pro-apoptotic drugs and for monitoring apoptosis in patient sera.

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Bivén, K; Erdal, H; Hägg, M; Ueno, T; Zhou, R; Lynch, M; Rowley, B; Wood, J; Zhang, C; Toi, M; Shoshan, M C; Linder, S

2003-06-01

We have developed an apoptosis assay based on measurement of a neoepitope of cytokeratin-18 (CK18-Asp396) exposed after caspase-cleavage and detected by the monoclonal antibody M30. The total amount of caspase-cleaved CK18 which has accumulated in cells and tissue culture media during apoptosis is measured by ELISA. The sensitivity is sufficient for use in the 96-well format to allow high-through-put screening of drug libraries. We here describe strategies allowing classification of pro-apoptotic compounds according to their profiles of induction of apoptosis in the presence of pharmacological inhibitors. The time course of induction of CK18 cleavage can furthermore be used to distinguish structurally similar compounds. We propose that compounds that induce rapid CK18 cleavage have mechanisms of actions distinct from conventional genotoxic and microtubuli-targeting agents, and we present one example of an agent that induces almost immediate mitochondrial depolarization and cytochrome c release. Finally, CK18-Asp396 cleavage products are released from cells in tissue culture, and presumably from tumor cells in vivo. These products can be measured in sera from cancer patients. We present evidence suggesting that it will be possible to use the M30-ELISA assay for measuring chemotherapy-induced apoptosis in patient sera, opening possibilities for monitoring therapy.

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)

Trypanosoma equiperdum Low Molecular Weight Proteins As Candidates for Specific Serological Diagnosis of Dourine

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Mirella Luciani

2018-03-01

Full Text Available The diagnosis of dourine can be difficult because the clinical signs of this disease in horses are similar to those of surra, caused by Trypanosoma evansi. Moreover, T. equiperdum and T. evansi are closely related and, so far, they cannot be distinguished using serological tests. In a previous work, the T. equiperdum protein pattern recognized by antibodies from dourine-infected horses and the humoral immune response kinetics were investigated by immunoblotting assay; a total of 20 sera from naturally and experimentally infected horses and from healthy animals were tested. Immunoblotting analysis showed that antibodies from infected horses specifically bind T. equiperdum low molecular weight proteins (from 16 to 35 kDa, which are not recognized by antibodies from uninfected horses. In this work, we tested other 615 sera (7 from naturally infected horses and 608 sera from healthy horses and donkeys: results confirmed the data obtained previously. In addition, six SDS-PAGE bands with molecular weight ranging from 10 to 37 kDa were analyzed by mass spectrometry, in order to identify immunogenic proteins that could be used as biomarkers for the diagnosis of dourine. A total of 167 proteins were identified. Among them, 37 were found unique for T. equiperdum. Twenty-four of them could represent possible candidate diagnostic antigens for the development of serological tests specific for T. equiperdum.

Microscopic agglutination test on captive rattlesnakes : Data on serovars and titers

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T.C.S. Rodrigues

2016-06-01

Full Text Available The microscopic agglutination test (MAT is considered the “golden standard” leptospirosis serodiagnostic test, but there is little information about it as it pertains to snakes. To fill this information gap, we provide data on serovars and titers of fifty-six Crotalus durissus collilineatus sera samples that tested positive by MAT (10.1016/j.actatropica.2016.02.006 (Rodrigues et al., 2016 [5]. These data are presented in a table, along with a description of the methodology used for sample collection and serologic testing.

Yellow fever virus envelope protein expressed in insect cells is capable of syncytium formation in lepidopteran cells and could be used for immunodetection of YFV in human sera

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Nagata Tatsuya

2011-05-01

Full Text Available Abstract Background Yellow fever is an haemorrhagic disease caused by a virus that belongs to the genus Flavivirus (Flaviviridae family and is transmitted by mosquitoes. Among the viral proteins, the envelope protein (E is the most studied one, due to its high antigenic potencial. Baculovirus are one of the most popular and efficient eukaryotic expression system. In this study a recombinant baculovirus (vSynYFE containing the envelope gene (env of the 17D vaccine strain of yellow fever virus was constructed and the recombinant protein antigenicity was tested. Results Insect cells infected with vSynYFE showed syncytium formation, which is a cytopathic effect characteristic of flavivirus infection and expressed a polypeptide of around 54 kDa, which corresponds to the expected size of the recombinant E protein. Furthermore, the recombinant E protein expression was also confirmed by fluorescence microscopy of vSynYFE-infected insect cells. Total vSynYFE-infected insect extracts used as antigens detected the presence of antibodies for yellow fever virus in human sera derived from yellow fever-infected patients in an immunoassay and did not cross react with sera from dengue virus-infected patients. Conclusions The E protein expressed by the recombinant baculovirus in insect cells is antigenically similar to the wild protein and it may be useful for different medical applications, from improved diagnosis of the disease to source of antigens for the development of a subunit vaccine.

Validation of pre-coated ELISA tests to detect antibodies against T. congolense and T. vivax

International Nuclear Information System (INIS)

Shumba, W.

2000-01-01

The anti-trypanosomal antibody detecting enzyme linked immunosorbent assay (ELISA) was first described in 1977 and was further developed for use in large scale surveys in Zimbabwe. More recently, the IAEA initiated a programme to improve the robustness and standardisation of the assay. The IAEA supplied plates pre-coated with either a crude T. congolense or T. vivax antigen and the reagents necessary for analysing samples. Parasitologically positive and negative sera were used to validate and determine the cut-off values of the two tests. The samples were tested and results analysed using a variety of cut-off values. The tests provided similar information although the T. congolense pre-coated plates gave significantly higher optical density values than the plates coated with T. vivax. Sensitivity and specificity values were calculated using the different cut-off points. Results indicate that the test using T. congolense antigen had the highest specificity and sensitivity for a given cut-off value. Although the test could distinguish positive from negative sera, it was quite difficult to provide a suitable cut-off value, but the value should be dictated by the use of the test. (author)

Primary cytomegalovirus infection in pregnant Egyptian women confirmed by cytomegalovirus IgG avidity testing.

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Kamel, N; Metwally, L; Gomaa, N; Sayed Ahmed, W A; Lotfi, M; Younis, S

2014-01-01

To determine the frequency of primary cytomegalovirus (CMV) infection in pregnant Egyptian women using CMV IgG avidity testing. A cross-sectional study was conducted at Suez Canal University Hospital, Ismailia, Egypt. A total of 546 pregnant women, presenting for routine antenatal screening, were tested for CMV IgG and IgM using a commercially available enzyme-linked immunosorbent assay (ELISA). Sera from CMV IgM-positive women were tested by CMV IgG avidity assay. All the 546 pregnant women were seropositive for anti-CMV IgG. Of the 546 women, 40 (7.3%) were positive or equivocal for IgM antibodies. All sera from the 40 women (IgG+/IgM+) showed a high or intermediate CMV IgG avidity index. Of the 40 women, 23 (57.5%) were in the second or third trimesters of pregnancy and had their first-trimester blood retrieved, and the tested CMV IgG avidity assay showed a high avidity index. Women who were IgM positive had no primary CMV infection in the index pregnancy as evidenced by the high CMV IgG avidity testing. © 2013 S. Karger AG, Basel.

Study of functional-performance deficits in athletes with previous ankle sprains

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hamid Babaee

2008-04-01

Full Text Available Abstract Background: Despite the importance of functional-performance deficits in athletes with history of ankle sprain few, studies have been carried out in this area. The aim of this research was to study relationship between previous ankle sprains and functional-performance deficits in athletes. Materials and methods: The subjects were 40 professional athletes selected through random sampling among volunteer participants in soccer, basketball, volleyball and handball teams of Lorestan province. The subjects were divided into 2 groups: Injured group (athletes with previous ankle sprains and healthy group (athletes without previous ankle sprains. In this descriptive study we used Functional-performance tests (figure 8 hop test and side hop test to determine ankle deficits and limitations. They participated in figure 8 hop test including hopping in 8 shape course with the length of 5 meters and side hop test including 10 side hop repetitions in course with the length of 30 centimeters. Time were recorded via stopwatch. Results: After data gathering and assessing information distributions, Pearson correlation was used to assess relationships, and independent T test to assess differences between variables. Finally the results showed that there is a significant relationship between previous ankle sprains and functional-performance deficits in the athletes. Conclusion: The athletes who had previous ankle sprains indicated functional-performance deficits more than healthy athletes in completion of mentioned functional-performance tests. The functional-performance tests (figure 8 hop test and side hop test are sensitive and suitable to assess and detect functional-performance deficits in athletes. Therefore we can use the figure 8 hop and side hop tests for goals such as prevention, assessment and rehabilitation of ankle sprains without spending too much money and time.

Plasma exchange to remove HIT antibodies: dissociation between enzyme-immunoassay and platelet activation test reactivities.

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Warkentin, Theodore E; Sheppard, Jo-Ann I; Chu, F Victor; Kapoor, Anil; Crowther, Mark A; Gangji, Azim

2015-01-01

Repeated therapeutic plasma exchange (TPE) has been advocated to remove heparin-induced thrombocytopenia (HIT) IgG antibodies before cardiac/vascular surgery in patients who have serologically-confirmed acute or subacute HIT; for this situation, a negative platelet activation assay (eg, platelet serotonin-release assay [SRA]) has been recommended as the target serological end point to permit safe surgery. We compared reactivities in the SRA and an anti-PF4/heparin IgG-specific enzyme immunoassay (EIA), testing serial serum samples in a patient with recent (subacute) HIT who underwent serial TPE precardiac surgery, as well as for 15 other serially-diluted HIT sera. We observed that post-TPE/diluted HIT sera-when first testing SRA-negative-continue to test strongly positive by EIA-IgG. This dissociation between the platelet activation assay and a PF4-dependent immunoassay for HIT antibodies indicates that patients with subacute HIT undergoing repeated TPE before heparin reexposure should be tested by serial platelet activation assays even when their EIAs remain strongly positive. © 2015 by The American Society of Hematology.

Differential reactivity of immune sera from human vaccinees with field strains of eastern equine encephalitis virus.

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Strizki, J M; Repik, P M

1995-11-01

Eastern equine encephalitis (EEE) virus is a mosquito-borne alphavirus that can produce a severe and often fatal acute encephalitis in humans, with significant neurologic sequelae in survivors. Due to the serious nature of the disease, an investigational inactivated EEE vaccine (PE-6) is available to individuals at risk for infection. Both serologic and recent molecular analyses of EEE viruses have demonstrated marked differences between the two antigenic varieties of EEE virus, designated North American (NA) and South American (SA). In view of these findings, we have examined the reactivity of sera from three individuals immunized with the EEE vaccine, derived from an NA isolate, with field strains of EEE virus. Anti-EEE serum antibodies from vaccinees reacted strongly in Western blot assays with both of the envelope (E1 and E2) glycoproteins of each NA strain examined, while reactivities with the glycoproteins of SA strains were substantially weaker and variable and dependent upon both the immune response of the vaccinee and the virus isolate assayed. Most striking was the modest to virtual lack of reactivity with the E2 protein of SA strains. Antigenic differences among the glycoproteins of EEE viruses were not as pronounced in immunoprecipitation analysis. Most significantly, although human immune sera displayed high neutralizing titers against each of the NA isolates examined, only negligible neutralizing titers were obtained against SA isolates. These data suggest that immunized individuals would mount an effective antibody response against infection with NA strains of EEE virus, but that further investigation is clearly warranted to fully assess the protective capability of the vaccine against infection with SA strains.

Immunoreactivity of the AAA+ chaperone ClpB from Leptospira interrogans with sera from Leptospira-infected animals.

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Krajewska, Joanna; Arent, Zbigniew; Więckowski, Daniel; Zolkiewski, Michal; Kędzierska-Mieszkowska, Sabina

2016-07-16

Leptospira interrogans is a spirochaete responsible for leptospirosis in mammals. The molecular mechanisms of the Leptospira virulence remain mostly unknown. Recently, it has been demonstrated that L. interrogans ClpB (ClpBLi) is essential for bacterial survival under stressful conditions and also during infection. The aim of this study was to provide further insight into the role of ClpB in L. interrogans and answer the question whether ClpBLi as a potential virulence factor may be a target of the humoral immune response during leptospiral infections in mammals. ClpBLi consists of 860 amino acid residues with a predicted molecular mass of 96.3 kDa and shows multi-domain organization similar to that of the well-characterized ClpB from Escherichia coli. The amino acid sequence identity between ClpBLi and E. coli ClpB is 52 %. The coding sequence of the clpB Li gene was cloned and expressed in E. coli BL21(DE3) strain. Immunoreactivity of the recombinant ClpBLi protein was assessed with the sera collected from Leptospira-infected animals and uninfected healthy controls. Western blotting and ELISA analysis demonstrated that ClpBLi activates the host immune system, as evidenced by an increased level of antibodies against ClpBLi in the sera from infected animals, as compared to the control group. Additionally, ClpBLi was found in kidney tissues of Leptospira-infected hamsters. ClpBLi is both synthesized and immunogenic during the infectious process, further supporting its involvement in the pathogenicity of Leptospira. In addition, the immunological properties of ClpBLi point to its potential value as a diagnostic antigen for the detection of leptospirosis.

Expression of Recombinant Streptokinase from Streptococcus Pyogenes and its Reaction with Infected Human and Murine Sera

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Molaee, Neda; Abtahi, Hamid; Mosayebi, Ghasem

2013-01-01

Objective(s): Streptokinase (SKa) is an antigenic protein which is secreted by Streptococcus pyogenes. Streptokinase induces inflammation by complement activation, which may play a role in post infectious diseases. In the present study, recombinant streptokinase from S. pyogenes was produced and showed that recombinant SKa protein was recognized by infected human sera using Western blot analysis. Materials and Methods: In this study, the ska gene from S. pyogenes was amplified and cloned into pET32a which is a prokaryotic expression vector. pET32a-ska was transformed to Escherichia coli BL21 (DE3) pLysS and gene expression was induced by IPTG. Protein production was improved by modification of composition of the bacterial culture media and altering the induction time by IPTG. The expressed protein was purified by affinity chromatography using the Ni-NTA resin. The integrity of the product was confirmed by Westernblot analysis using infected mice. Serum reactivity of five infected individuals was further analyzed against the recombinant SKa protein. Results: Data indicated that recombinant SKa protein from S. pyogenes can be recognized by patient and mice sera. The concentration of the purified recombinant protein was 3.2 mg/L of initial culture. The highest amount of the expressed protein after addition of IPTG was obtained in a bacterial culture without glucose with the culture optical density of 0.8 (OD600 = 0.8). Conclusion : Present data shows, recombinant SKa protein has same epitopes with natural form of this antigen. Recombinant SKa also seemed to be a promising antigen for the serologic diagnosis of S. pyogenes infections. PMID:24171077

Expression of Recombinant Streptokinase from Streptococcus Pyogenes and Its Reaction with Infected Human and Murine Sera

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Neda Molaee

2013-09-01

Full Text Available   Objective(s: Streptokinase (SKa is an antigenic protein which is secreted by Streptococcus pyogenes. Streptokinase induces inflammation by complement activation, which may play a role in post infectious diseases. In the present study, recombinant streptokinase from S. pyogenes was produced and showed that recombinant SKa protein was recognized by infected human sera using Western blot analysis.   Materials and Methods: In this study, the ska gene from S. pyogenes was amplified and cloned into pET32a which is a prokaryotic expression vector. pET32a-ska was transformed to Escherichia coli BL21 (DE3 pLysS and gene expression was induced by IPTG. Protein production was improved by modification of composition of the bacterial culture media and altering the induction time by IPTG. The expressed protein was purified by affinity chromatography using the Ni-NTA resin. The integrity of the product was confirmed by Westernblot analysis using infected mice. Serum reactivity of five infected individuals was further analyzed against the recombinant SKa protein. Results: Data indicated that recombinant SKa protein from S. pyogenes can be recognized by patient and mice sera. The concentration of the purified recombinant protein was 3.2 mg/L of initial culture. The highest amount of the expressed protein after addition of IPTG was obtained in a bacterial culture without glucose with the culture optical density of 0.8 (OD600 = 0.8. Conclusion : Present data shows, recombinant SKa protein has same epitopes with natural form of this antigen. Recombinant SKa also seemed to be a promising antigen for the serologic diagnosis of S. pyogenes infections.

Autoantibodies from primary biliary cirrhosis patients with anti-p95c antibodies bind to recombinant p97/VCP and inhibit in vitro nuclear envelope assembly

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MIYACHI, K; HIRANO, Y; HORIGOME, T; MIMORI, T; MIYAKAWA, H; ONOZUKA, Y; SHIBATA, M; HIRAKATA, M; SUWA, A; HOSAKA, H; MATSUSHIMA, S; KOMATSU, T; MATSUSHIMA, H; HANKINS, R W; FRITZLER, M J

2004-01-01

We have reported previously that p95c, a novel 95-kDa cytosolic protein, was the target of autoantibodies in sera of patients with autoimmune hepatic diseases. We studied 30 sera that were shown previously to immunoprecipitate a 95 kDa protein from [35S]-methionine-labelled HeLa lysates and had a specific precipitin band in immunodiffusion. Thirteen sera were available to test the ability of p95c antibodies to inhibit nuclear envelope assembly in an in vitro assay in which confocal fluorescence microscopy was also used to identify the stages at which nuclear assembly was inhibited. The percentage inhibition of nuclear envelope assembly of the 13 sera ranged from 7% to 99% and nuclear envelope assembly and the swelling of nucleus was inhibited at several stages. The percentage inhibition of nuclear assembly was correlated with the titre of anti-p95c as determined by immunodiffusion. To confirm the identity of this autoantigen, we used a full-length cDNA of the p97/valosin-containing protein (VCP) to produce a radiolabelled recombinant protein that was then used in an immunoprecipitation (IP) assay. Our study demonstrated that 12 of the 13 (93%) human sera with antibodies to p95c immunoprecipitated recombinant p97/VCP. Because p95c and p97 have similar molecular masses and cell localization, and because the majority of sera bind recombinant p97/VCP and anti-p95c antibodies inhibit nuclear assembly, this is compelling evidence that p95c and p97/VCP are identical. PMID:15147362

Determination of IgE antibodies to Polistes dominulus, Vespula germanica and Vespa crabro in sera of patients allergic to vespids.

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Blanca, M; Garcia, F; Miranda, A; Carmona, M J; Garcia, J; Fernandez, J; Terrados, S; Vega, J M; Juarez, C

1991-02-01

The study was undertaken to investigate the presence of IgE antibodies to Polistes dominulus (PD), Vespula germanica (VG) and Vespa crabro (VC) in a large group of sera belonging to patients sensitized to Vespids in Spain. RAST values showed that although the majority of patients had IgE antibodies to PD, VG and VC, there was a marked predominance of PD. These results were related to the distribution of the insect in the areas where the sera were obtained. Due to geographical and insect distribution differences, the whole area was divided into three zones: Central, East and South. Comparison of the positive RAST values obtained indicated that, although the positivity to PD predominated over VG and this over VC, there were significant differences in percentage positivities to each vespid in the different regions studied. The results of the RAST absorption studies indicated that in most instances patients were originally sensitized to one vespid and were RAST positive to the other venoms due to cross-reactivity. Only in a minority of cases were coexisting antibodies to two insects present. These results show that PD and VG are the important vespids followed to a lesser extent by VC. This study provides relevant information concerning insect distribution sensitivity in a European country.

Detection of antibodies against H5 and H7 strains in birds: evaluation of influenza pseudovirus particle neutralization tests

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Sofie Wallerström

2014-01-01

Full Text Available Introduction: Avian influenza viruses circulate in bird populations, and it is important to maintain and uphold our knowledge of the viral strains that are currently of interest in this context. Here, we describe the use of hemagglutinin-pseudotype retroviruses based on highly pathogenic influenza viruses for the screening of avian sera for influenza A antibodies. Our aim was also to determine whether the pseudovirus neutralization tests that we assessed were sensitive and simple to use compared to the traditional methods, including hemagglutination inhibition assays and microneutralization tests. Material and methods: H5 and H7 pseudovirus neutralization tests were evaluated by using serum from infected rabbits. Subsequently, the assays were further investigated using a panel of serum samples from avian species. The panel contained samples that were seropositive for five different hemagglutinin subtypes as well as influenza A seronegative samples. Results and discussion: The results suggest that the pseudovirus neutralization test is an alternative to hemagglutination inhibition assays, as we observed comparable titers to those of both standard microneutralizations assays as well as hemagglutinin inhibition assays. When evaluated by a panel of avian sera, the method also showed its capability to recognize antibodies directed toward low-pathogenic H5 and H7. Hence, we conclude that it is possible to use pseudoviruses based on highly pathogenic avian influenza viruses to screen avian sera for antibodies directed against influenza A subtypes H5 and H7.

Application of the electromagnetic borehole flowmeter and evaluation of previous pumping tests at Paducah Gaseous Diffusion Plant. Final report, June 15, 1992--August 31, 1992

Energy Technology Data Exchange (ETDEWEB)

Young, S.C.; Julian, S.C.; Neton, M.J.

1993-01-01

Multi-well pumping tests have been concluded at wells MW79, MW108, and PW1 at the Paducah Gaseous Diffusion Plant (PGDP) to determine the hydraulic properties of the Regional Gravel Aquifer (RGA). Soil cores suggest that the RGA consists of a thin sandy facies (2 to 6 feet) at the top of a thicker (> 10 feet) gravelly facies. Previous analyses have not considered any permeability contrast between the two facies. To assess the accuracy of this assumption, TVA personnel conducted borehole flowmeter tests at wells MW108 and PW1. Well MW79 could not be tested. The high K sand unit is probably 10 times more permeable than comparable zone in the gravelly portion of the RGA. Previous analyses of the three multi-well aquifer tests do not use the same conceptual aquifer model. Data analysis for one pumping test assumed that leakance was significant. Data analysis for another pumping test assumed that a geologic boundary was significant. By collectively analyzing all three tests with the borehole flowmeter results, the inconsistency among the three pumping tests can be explained. Disparity exists because each pumping test had a different placement of observation wells relative to the high K zone delineating by flowmeter testing.

Alpha subunit of glycoprotein hormones in the sera of acromegalic patients and its mRNA in the tumors.

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Machiavelli, G A; Artese, R; Benencia, H; Bruno, O; Guerra, L; Basso, A; Burdman, J A

1999-04-01

Within a population of 16 pituitary adenomas we found high levels of glycoprotein alpha subunits in the sera of patients with somatotrophic tumors. This finding was correlated with the presence of mRNA alpha subunit in these tumors indicating the adenomas themselves as the origin of the circulating alpha-subunit. Synthesis of these two hormones, which are chemically very different, by the same tumor cells indicates a high degree of differentiation of these cells. We are unable at this time to conclusively correlate differentiation of these tumors aggressively.

A Novel Tool for High-Throughput Screening of Granulocyte-Specific Antibodies Using the Automated Flow Cytometric Granulocyte Immunofluorescence Test (Flow-GIFT

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Xuan Duc Nguyen

2011-01-01

Full Text Available Transfusion-related acute lung injury (TRALI is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT and granulocyte agglutination test (GAT were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti—HNA 3a, n = 3; anti—HNA-1b, n = 1 and GAT (anti—HNA-2a, n = 1. The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of

A novel tool for high-throughput screening of granulocyte-specific antibodies using the automated flow cytometric granulocyte immunofluorescence test (Flow-GIFT).

Science.gov (United States)

Nguyen, Xuan Duc; Dengler, Thomas; Schulz-Linkholt, Monika; Klüter, Harald

2011-02-03

Transfusion-related acute lung injury (TRALI) is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT) has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti-HNA 3a, n = 3; anti-HNA-1b, n = 1) and GAT (anti-HNA-2a, n = 1). The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of blood products.

Colostral whey concentrate supplement increases complement activity in the sera of neonatal calves.

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Rokka, S; Korhonen, B H; Nousiainen, J; Marnila, P

2001-08-01

We evaluated the effect of a commercial bovine colostral whey on the complement-mediated immune responses of calves. Two groups of neonatal calves were fed, in addition to whole milk (WM) and pooled colostrum (PC), different amounts of a commercial immunoglobulin concentrate made from pooled colostral whey (Ig-C) for the first two feedings post natum. The control group was fed WM and PC only. Serum samples were obtained at the ages of 2, 7, 14 and 30 d. Bacteriolytic activity against complement-sensitive Escherichia coli JM103 and opsonic activity against complement-lysis-resistant E. coli IH3080 strains were studied, as well as the levels of C3 complement component and E. coli JM103 specific antibodies in the sera. Groups fed Ig-C had 2-3 times higher bacteriolytic activity than the control group of both the classic (P complement activities of serum can be increased substantially by feeding colostral whey concentrate to calves during their first days of life.

Catalase activity of IgG antibodies from the sera of healthy donors and patients with schizophrenia.

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Ermakov, Evgeny A; Smirnova, Ludmila P; Bokhan, Nikolay A; Semke, Arkadiy V; Ivanova, Svetlana A; Buneva, Valentina N; Nevinsky, Georgy A

2017-01-01

We present first evidence showing that some electrophoretically homogeneous IgGs from the sera of patients with schizophrenia (36.4%) and their Fab and F(ab)2 fragments as well as from healthy donors (33.3%) possess catalase activity. The relative catalase activity of IgGs from the sera of individual schizophrenia patients (and healthy donors) significantly varied from patient to patient, but the activity of IgGs from healthy donors is on average 15.8-fold lower than that for schizophrenia patients. After extensive dialysis of purified IgGs against EDTA chelating metal ions, the relative catalase activity of IgGs decreases on average approximately 2.5-3.7-fold; all IgGs possess metal-dependent and independent catalase activity. The addition of external Me2+ ions to dialyzed and non-dialyzed IgGs leads to a significant increase in their activity. The best activator of dialyzed and non-dialyzed IgGs is Co2+, the activation by Cu2+, Mn2+, and Ni2+ ions were rare and always lower than by Co2+. Every IgG preparation demonstrates several individual sets of very well expressed pH optima in the pH range from 4.0 to 9.5. These data speak for the individual repertoire of catalase IgGs in every person and an extreme diversity of abzymes in their pH optima and activation by different metal ions. It is known that antioxidant enzymes such as superoxide dismutases, catalases, and glutathione peroxidases represent critical defense mechanisms preventing oxidative modifications of DNA, proteins, and lipids. Catalase activity of human IgGs could probably also play a major role in the protection of organisms from oxidative stress and toxic compounds.

In-gel detection of esterase-like albumin activity: Characterization of esterase-free sera albumin and its putative role as non-invasive biomarker of hepatic fibrosis

Directory of Open Access Journals (Sweden)

Areeba Ahmad

2017-07-01

Full Text Available Albumin is a globular and un-glycosylated multifunctional plasma protein and thus correlated with several human diseases. Owing to esterase contamination, albumin levels are usually misleading. In this study, we propose methodical accuracy for albumin estimation taking healthy and fibrotic rats. Liver fibrosis in rats was generated by N′-Nitrosodimethylamine (NDMA (10 mg/kg body weight within three weeks followed by its confirmation through H&E and immunohistochemical staining for α-SMA expression. Animal sera were screened by native polyacrylamide gel electrophoresis (native-PAGE (7.5%. In-gel esterase-like albumin activity was detected using α- and β-naphthyl acetate (5.58 × 10−3 mM; pH 7.5 as substrate. Sera albumin was purified from unstained PA gel-slices through electroelution. Subsequent to conformation of albumin purity by its molecular weight determination using SDS–PAGE (10% and peptide mass fingerprinting by MALDI-TOF-MS, samples were treated with different concentrations of urea. Urea-treated albumins were screened for esterase activity, conformational change and, albumin levels by immunoblotting. Our results demonstrate that esterase-like albumin activity in rat sera albumin is located in domain-III. The esterase-like activity remains detectable up to 4 M urea, which diminishes with increasing urea concentrations. Further, immunoblotting of urea-treated albumin samples displays a significant decline in purified protein bands, indicating hypoalbuminemia during hepatic fibrosis in rats. In conclusion, the present approach of albumin separation and estimation is of potential interest and may be recommended for diagnostic purposes.

A serosurvey for ruminant pestivirus exposure conducted using cattle sera collected for brucellosis surveillance in the United States.

Science.gov (United States)

Bauermann, Fernando V; Ridpath, Julia F; Dargatz, David A

2017-01-01

Four species of ruminant pestivirus are currently circulating in the United States: Bovine viral diarrhea virus 1 and 2 (BVDV-1, -2; predominant host: cattle), Border disease virus (BDV; predominant host: sheep), and pronghorn virus (sporadically detected in wild ruminants). A third bovine pestivirus called HoBi-like virus has been detected in cattle and water buffalo in South America, Asia, and Europe. To date, no isolations of HoBi-like viruses from U.S. cattle have been reported. To assess exposure, 2,000 cattle sera, collected between 2014 and 2015 as part of the U.S. brucellosis surveillance program, were tested for antibodies against BVDV-1, BVDV-2, and HoBi-like viruses. In addition, RNA was extracted and tested by reverse transcription-polymerase chain reaction for the presence of pestiviruses; all samples tested negative. The percent of VN-positive samples was 91.3% for BVDV-1, 89.3% for BVDV-2, and 84.9% for HoBi-like viruses. Because the 3 bovine pestiviruses are antigenically cross-reactive, the comparative level of antibody against each pestivirus species was determined. Based on comparative titers, samples were segregated into 6 categories: no titers (7.6%), titers clearly higher against BVDV-1 (22.2%), titers substantially higher against BVDV-2 (9.1%), BVDV-1 and BVDV-2 titers equivalent but substantially higher than HoBi titers (25.7%), titers substantially higher against HoBi-like viruses (0%), and equivocal (35.4%). Titers tended to be higher against BVDV-1 than BVDV-2. However, the overall percentage of animals with titers below levels considered protective against acute bovine pestivirus infection were ~11% for BVDV-1, 12% BVDV-2, and 18% for HoBi-like virus.

Recombinant major urinary proteins of the mouse in specific IgE and IgG testing

NARCIS (Netherlands)

Krop, Esmeralda J. M.; Matsui, Elizabeth C.; Sharrow, Scott D.; Stone, Martin J.; Gerber, Peter; van der Zee, Jaring S.; Chapman, Martin D.; Aalberse, Rob C.

2007-01-01

BACKGROUND: Recombinant allergens are preferred over natural allergen extracts in measuring antibodies. We tested the use of recombinant variants of the major mouse allergen Mus m 1 in detection of mouse-specific antibodies in sera of laboratory animal workers and children. METHODS: Six recombinant

Zika virus and blood transfusion: the experience of French Polynesia.

Science.gov (United States)

Bierlaire, Damien; Mauguin, Sylvie; Broult, Julien; Musso, Didier

2017-03-01

Between October 2013 and March 2014, French Polynesia experienced the largest Zika virus (ZIKV) outbreak ever described before the emergence of ZIKV in the Americas in 2015. As arbovirus transfusion-transmitted (TT) infections have been previously reported, we hypothesized that transfusion of blood products could also transmit ZIKV. Mitigation strategies to prevent ZIKV-TT infections included nonspecific measures and the implementation of a laboratory developed ZIKV-specific nucleic acid testing (NAT) assay. Donor sera were tested in pools of 3 and constitutive sera of ZIKV-reactive pools were tested individually. Donor sera were tested prospectively and retrospectively. A posttransfusion follow-up of a patient transfused with ZIKV RNA-reactive blood products was implemented. NAT detected 42 blood donor sera as ZIKV RNA reactive of 1505 tested (2.8%). Thirty ZIKV RNA-reactive blood products collected before the implementation of NAT were transfused to 26 recipients. Posttransfusion investigations were conducted by the hemovigilance unit and data were available for 12 recipients. Symptomatic ZIKV-TT infections were not reported. Predonation screening of blood donors, postdonation information, products discard, and quarantine of blood products were not effective enough to prevent transfusion of ZIKV RNA-reactive blood products. ZIKV NAT was an effective measure once implemented to prevent transfusion of ZIKV RNA-reactive blood products but it is difficult to evaluate the effectiveness of this measure to prevent ZIKV-TT infection, which is a rare event. © 2017 AABB.

[Serological study of leptospirosis in equids, camelids and bovids from Djibouti].

Science.gov (United States)

Roqueplo, C; Davoust, B; Mulot, B; Lafrance, B; Kodjo, A

2011-10-01

Sera obtained from 31 domestic and feral animals in Djibouti were assayed for leptospiral antibodies using the microscopic agglutination test. Antibodies were detected in 26 samples (84%), corresponding to 116 positive reactions. The most common antigen serogroups were Icterohaemorrhagiae and Australis. The highest titre was recorded for serovar Munchen (1:1280) in sera from Somalian wild asses and goats. This study shows a broad dispersion and high prevalence of the different Leptospira serogroups tested. High biodiversity has been previously reported in tropical countries and is thought to be linked to the wide range of reservoir mammals. Additional study will be needed to identify the reservoirs of the different serogroups in this part of Africa.

A macroecological analysis of SERA derived forest heights and implications for forest volume remote sensing.

Science.gov (United States)

Brolly, Matthew; Woodhouse, Iain H; Niklas, Karl J; Hammond, Sean T

2012-01-01

Individual trees have been shown to exhibit strong relationships between DBH, height and volume. Often such studies are cited as justification for forest volume or standing biomass estimation through remote sensing. With resolution of common satellite remote sensing systems generally too low to resolve individuals, and a need for larger coverage, these systems rely on descriptive heights, which account for tree collections in forests. For remote sensing and allometric applications, this height is not entirely understood in terms of its location. Here, a forest growth model (SERA) analyzes forest canopy height relationships with forest wood volume. Maximum height, mean, H₁₀₀, and Lorey's height are examined for variability under plant number density, resource and species. Our findings, shown to be allometrically consistent with empirical measurements for forested communities world-wide, are analyzed for implications to forest remote sensing techniques such as LiDAR and RADAR. Traditional forestry measures of maximum height, and to a lesser extent H₁₀₀ and Lorey's, exhibit little consistent correlation with forest volume across modeled conditions. The implication is that using forest height to infer volume or biomass from remote sensing requires species and community behavioral information to infer accurate estimates using height alone. SERA predicts mean height to provide the most consistent relationship with volume of the height classifications studied and overall across forest variations. This prediction agrees with empirical data collected from conifer and angiosperm forests with plant densities ranging between 10²-10⁶ plants/hectare and heights 6-49 m. Height classifications investigated are potentially linked to radar scattering centers with implications for allometry. These findings may be used to advance forest biomass estimation accuracy through remote sensing. Furthermore, Lorey's height with its specific relationship to remote sensing

Allergen-specific IgE testing in the diagnosis of food allergy and the event of a positive match in the bioinformatics search

DEFF Research Database (Denmark)

van Ree, Ronald; Vieths, Stefan; Poulsen, Lars K.

2006-01-01

Current documents on risk assessment of genetically modified foods recommend including IgE-binding tests on sera from allergic patients. However, there is no generally accepted recommendation on technical aspects of the testing procedures or on the interpretation of the results, despite that fact...

Anti-idiotypic antibodies to poliovirus antibodies in commercial immunoglubulin preparations, human serum and milk.

NARCIS (Netherlands)

M. Hahn-Zoric; B. Carlsson; S. Jeansson; H.P. Ekre; A.D.M.E. Osterhaus (Albert); D. Roberton; L.A. Hanson

1993-01-01

textabstractOur previous studies have suggested that fetal antibody production can be induced by maternal antiidiotypic antibodies transferred to the fetus via the placenta. We tested commercial Ig, sera, and milk for the presence of anti-idiotypic antibodies to poliovirus type 1, using affinity

Isolation, purification and partial characterization of early pregnancy factor (EPF) from sera of pregnant women.

Science.gov (United States)

Haq, A; Mothi, B A; Al-Hussein, K; Al-Tufail, M; Hollanders, J; Jaroudi, K; Al-Waili, N; Shabani, M

2001-05-29

Early pregnancy factor (EPF) is a pregnancy protein, which is secreted into the maternal serum 12-16 hours after fertilization. It is thought to be an immunosuppressive molecule. EPF is detected in pregnant woman's serum by the rosette inhibition assay (RIA). In this study, EPF was purified from the pregnant woman's sera by using ion exchange chromatography and analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins which showed a positive result with the RIA, were found to be 35 kDa and 17 kDa molecular weights. The biological activities of these proteins were stable upon heat treatment at 56 degrees C for 30 min. Proteins isolated and purified in this study might be of great significance to the field of human reproduction with particular reference to pregnancy and recurrent abortion.

Deficiency of the Chemotactic Factor Inactivator in Human Sera with α1-Antitrypsin Deficiency

Science.gov (United States)

Ward, Peter A.; Talamo, Richard C.

1973-01-01

As revealed by appropriate fractionation procedures, human serum deficient in α1-antitrypsin (α1-AT) is also deficient in the naturally occurring chemotactic factor inactivator. These serum donors had severe pulmonary emphysema. Serum from patients with clinically similar pulmonary disease, but with presence of α1-AT in the serum, showed no such deficiency of the chemotactic factor inactivator. When normal human serum and α1-AT-deficient human sera are chemotactically activated by incubation with immune precipitates, substantially more chemotactic activity is generated in α1-AT-deficient serum. These data indicate that in α1-AT-deficient serum there is an imbalance in the generation and control of chemotactic factors. It is suggested that the theory regarding development of pulmonary emphysema in patients lacking the α1-antitrypsin in their serum should be modified to take into account a deficiency of the chemotactic factor inactivator. PMID:4683887

Long- and short-term retention of traditional instruction vs. previously tested tactual vs. innovative tactual resources on the achievement and attitudes of second-grade students in science

Science.gov (United States)

Mitchell, Sherese A.

This researcher investigated the long- and short-term retention of information using traditional instruction versus previously tested tactual resources versus innovative tactual resources on the achievement and attitudes of second-grade students in science. The processing of new and difficult knowledge has challenged many young children who tend to be kinesthetic or tactual learners. In compliance with the National Science Education Standards, students should be actively engaged in their own learning. Therefore, to boost student achievement in science, the use of tactual materials was implemented. The sample included 67 second-grade students drawn from three heterogeneously grouped classes in a low socio-economic neighborhood. It consisted of 30 females and 37 males of which 97 percent were African American, 2 percent were Hispanic, and 1 percent Other. Students were unaware of their diagnosed learning-style preference(s) during the instruction and assessment phases of the study. Therefore, students' knowledge of their learning-style preferences could not have had any impact on their achievement or attitudes. A counterbalanced research design was employed. During the first session, Group 1 was taught with previously tested tactual resources (Electroboards, Flip Chutes, Fact Wheels, and Fact Fans), and Group 3 was taught traditionally. During the second session of instruction, Group 1 received instruction with innovative tactual resources, Group 2 received traditional instruction, Group 3 received instruction with previously tested tactual resources. During the final session of instruction, Group 1 received traditional instruction, Group 2 received instruction with previously tested tactual resources, and Group 3 received instruction with innovative tactual resources. The results indicated that the use of tactual materials, regardless of whether they were previously tested or innovative, produced higher achievement gains and more positive attitudes than traditional

Evaluation of a rapid immunochromatographic dipstick test for detection of antibodies to Trypanosoma cruzi in dogs experimentally infected with isolates obtained from opossums (Didelphis virginiana), armadillos (Dasypus novemcinctus), and dogs (Canis familiaris) from the United States.

Science.gov (United States)

Rosypal, Alexa C; Hill, Roderick; Lewis, Samantha; Barr, Stephen C; Valadas, Samantha; Gennari, Solange Maria; Lindsay, David S

2011-02-01

Dogs are reservoir hosts for Trypanosoma cruzi , the causative agent of American trypanosomiasis. A rapid immunochromatographic dipstick test (ICT) is available commercially for canine serological testing. The ICT was developed with the use of sera from South American dogs, but it is not routinely used in the United States. We evaluated the utility of the ICT in detecting anti-T. cruzi antibodies in dogs from the United States. Dogs (N  =  64) were experimentally infected with United States' isolates of T. cruzi from an opossum (Didelphis virginiana), an armadillo (Dasypus novemcinctus), and a domestic dog (Canis familiaris), and were tested after experimental infection. Sera from uninfected United States dogs (n  =  79; hemaculture negative) were used as negative controls. In a blind study, sera were tested by the ICT and compared to the indirect immunofluorescent antibody test with the use of Brazil-strain epimastigotes as antigen. The sensitivity of the ICT was 91% and the specificity was 98% in dogs experimentally infected with United States isolates. Our study indicates that the ICT could be a useful screening tool for serological surveillance of canine T. cruzi exposure in the United States.

Sera from Preeclampsia Patients Elicit Symptoms of Human Disease in Mice and Provide a Basis for an in Vitro Predictive Assay

OpenAIRE

Kalkunte, Satyan; Boij, Roland; Norris, Wendy; Friedman, Jennifer; Lai, Zhongbin; Kurtis, Jonathan; Lim, Kee-Hak; Padbury, James F.; Matthiesen, Leif; Sharma, Surendra

2010-01-01

Early diagnosis and treatment of preeclampsia would significantly reduce maternal and fetal morbidity and mortality. However, its etiology and prediction have remained elusive. Based on the hypothesis that sera from patients with preeclampsia could function as a “blueprint” of causative factors, we describe a serum-based pregnancy-specific mouse model that closely mirrors the human condition as well as an in vitro predictive assay. We show that a single administration of human preeclampsia se...

Near-infrared Raman spectroscopy to detect anti-Toxoplasma gondii antibody in blood sera of domestic cats: quantitative analysis based on partial least-squares multivariate statistics

Science.gov (United States)

Duarte, Janaína; Pacheco, Marcos T. T.; Villaverde, Antonio Balbin; Machado, Rosangela Z.; Zângaro, Renato A.; Silveira, Landulfo

2010-07-01

Toxoplasmosis is an important zoonosis in public health because domestic cats are the main agents responsible for the transmission of this disease in Brazil. We investigate a method for diagnosing toxoplasmosis based on Raman spectroscopy. Dispersive near-infrared Raman spectra are used to quantify anti-Toxoplasma gondii (IgG) antibodies in blood sera from domestic cats. An 830-nm laser is used for sample excitation, and a dispersive spectrometer is used to detect the Raman scattering. A serological test is performed in all serum samples by the enzyme-linked immunosorbent assay (ELISA) for validation. Raman spectra are taken from 59 blood serum samples and a quantification model is implemented based on partial least squares (PLS) to quantify the sample's serology by Raman spectra compared to the results provided by the ELISA test. Based on the serological values provided by the Raman/PLS model, diagnostic parameters such as sensitivity, specificity, accuracy, positive prediction values, and negative prediction values are calculated to discriminate negative from positive samples, obtaining 100, 80, 90, 83.3, and 100%, respectively. Raman spectroscopy, associated with the PLS, is promising as a serological assay for toxoplasmosis, enabling fast and sensitive diagnosis.

A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss

Science.gov (United States)

Minor, Kirsty L.; Anderson, Victoria L.; Davis, Katie S.; Van Den Berg, Albert H.; Christie, James S.; Löbach, Lars; Faruk, Ali Reza; Wawra, Stephan; Secombes, Chris J.; Van West, Pieter

2014-01-01

Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. PMID:25088077

Overlapping but distinct specificities of anti-liver-kidney microsome antibodies in autoimmune hepatitis type II and hepatitis C revealed by recombinant native CYP2D6 and novel peptide epitopes

Science.gov (United States)

Klein, R; Zanger, U M; Berg, T; Hopf, U; Berg, P A

1999-01-01

Anti-liver-kidney microsome antibodies (anti-LKM) occur in autoimmune hepatitis (AIH) type II and in a subset of patients with hepatitis C. Anti-LKM1 in AIH are directed against cytochrome P4502D6 (CYP2D6), but conflicting data exist concerning the specificity of anti-LKM in hepatitis C. The aim of this study was to evaluate binding specificities of anti-LKM antibodies in both diseases using novel test antigens as well as their inhibitory capacity on CYP2D6 enzyme activity. Sera from 22 patients with AIH type II and 17 patients with hepatitis C being anti-LKM-positive in the immunofluorescence test were investigated for binding to native recombinant CYP2D6 and liver microsomes by ELISA and immunoblotting, and to synthetic peptides covering the region 254–339 (254–273, 257–269, 270–294, 291–310, 307–324, 321–339, 373–389) as well as the novel peptide 196–218 by ELISA. Furthermore, all sera were tested for inhibition of CYP2D6-dependent bufuralol 1′-hydroxylase activity. Twenty of the 22 AIH type II sera (91%) and nine of the 17 hepatitis C sera (53%) were positive for CYP2D6 by ELISA and/or immunoblotting. The previously described major peptide epitope comprising CYP2D6 amino acids 257–269 was recognized by 16 of the 22 AIH sera but by only one hepatitis C serum. A further epitope, 196–218, could be defined for the first time as another immunodominant epitope for AIH because it was recognized by 15 of the 22 AIH (68%) but only three of the 17 hepatitis C sera (18%). With the exception of the peptide 254–273, the other peptides showed no significant reactivity. Analysing the inhibitory properties of anti-LKM antibodies it emerged that 95% of AIH sera and 88% of hepatitis C sera inhibited enzyme function. These data indicate that anti-LKM antibodies in AIH and hepatitis C react with CYP2D6, as shown by their inhibitory activity, and that besides the known epitope 257–269 a further immunodominant epitope exists on CYP2D6 which is recognized

Simultaneous determination of dihydrotestosterone and its metabolites in mouse sera by LC-MS/MS with chemical derivatization.

Science.gov (United States)

Gorityala, Shashank; Yang, Shuming; Montano, Monica M; Xu, Yan

2018-07-15

Androgens play a vital role in prostate cancer development, and their elimination and blockade are essential in the disease management. DHT is the key ligand for androgen receptor (AR) in the prostate. It is locally synthesized from testosterone. In the prostate, DHT is predominantly metabolized to α-diol and β-diol. Recent studies indicate that impaired DHT catabolism is associated with prostate cancer, signifying the necessity of a sensitive quantitative method for the determination of DHT and its metabolites. In this work, an LC-MS/MS method for the simultaneous quantification of DHT and its metabolites was developed and validated. Steroid-free sera were prepared and used for the preparation of sera calibrators and quality controls (QCs). DHT and its metabolites along with their respective stable heavy isotope labeled analytes representing internal standards were first extracted with methyl tertiary-butyl ether (MTBE) and derivatized with picolinic acid (PA). The derivatized analytes were then extracted again with MTBE, dried under nitrogen and reconstituted in the mobile phase (80% methanol and 0.2% formic acid in water). Baseline chromatographic separation of the derivatized analytes was achieved isocratically on XTerra C18 column (2.1 × 100 mm) using the mobile phase at a flow rate of 0.25 mL/min. Quantitation was performed using multiple-reaction-monitoring mode with positive electrospray ionization. The method has calibration ranges from 0.0500 ng/mL to 50.0 ng/mL for DHT and its two metabolites with acceptable assay precision, accuracy, recovery, and matrix factor. It was applied to the determination of DHT and its metabolites in an animal study. Copyright © 2018 Elsevier B.V. All rights reserved.

A multi-analyte biosensor for the simultaneous label-free detection of pathogens and biomarkers in point-of-need animal testing.

Science.gov (United States)

Ewald, Melanie; Fechner, Peter; Gauglitz, Günter

2015-05-01

For the first time, a multi-analyte biosensor platform has been developed using the label-free 1-lambda-reflectometry technique. This platform is the first, which does not use imaging techniques, but is able to perform multi-analyte measurements. It is designed to be portable and cost-effective and therefore allows for point-of-need testing or on-site field-testing with possible applications in diagnostics. This work highlights the application possibilities of this platform in the field of animal testing, but is also relevant and transferable to human diagnostics. The performance of the platform has been evaluated using relevant reference systems like biomarker (C-reactive protein) and serology (anti-Salmonella antibodies) as well as a panel of real samples (animal sera). The comparison of the working range and limit of detection shows no loss of performance transferring the separate assays to the multi-analyte setup. Moreover, the new multi-analyte platform allows for discrimination between sera of animals infected with different Salmonella subtypes.

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.

Development of a solid-phase radioimmunoassay for antibody to antigens of Babesia bovis infected bovine erythrocytes

Energy Technology Data Exchange (ETDEWEB)

Kahl, L.P.; Anders, R.F.; Mitchell, G.F. (Walter and Eliza Hall Inst. of Medical Research, Parkville (Australia)); Callow, L.L.; Rodwell, B.J. (Animal Research Inst. Wacol (Australia). Tick Fever Research Centre)

1982-06-01

A quantitative solid-phase radioimmunoassay (RIA) has been developed for the purpose of ranking sera from exposed animals according to their titre of anti-B. bovis antibody. The antigen was a sonicate of lysed infected blood cells, and antibody binding was detected with /sup 125/I-labelled anti-bovine IgG. The assay was tested using sera from experimentally infected splenectomized and intact cattle and from animals resident in an endemic area. A high specificity for B. bovis was demonstrated. There was good agreement in identifying exposed cattle when the test was compared with an indirect fluorescent antibody (IFA) test although no correlation was seen between titres obtained in the two tests. Analysis of the effects of challenge infection with B. bovis on IFA and RIA titres in previously exposed animals illustrated that the RIA was a more sensitive test for detecting changes in antibody titre.

Evaluation of two automated chemiluminescence immunoassays, the LIAISON Treponema Screen and the ARCHITECT Syphilis TP, and the Treponema pallidum particle agglutination test for laboratory diagnosis of syphilis.

Science.gov (United States)

Wellinghausen, Nele; Dietenberger, Hanna

2011-08-01

Automated Treponema pallidum-specific chemi-luminescence immunoassays (CLIA) run on random-access analyzers allow for rapid diagnosis of syphilis infection. We evaluated the LIAISON Treponema Screen (LIA) and the ARCHITECT Syphilis TP (ARCH) in comparison to the Treponema pallidum particle agglutination (TPPA) test, as a screening test for syphilis. We performed a prospective study using 577 sera submitted for diagnosis of syphilis, including 318 samples from pregnant women. In addition, 42 stored sera from 32 patients with clinically and serologically characterized syphilis infection were investigated. In the prospective study, the sensitivity and specificity of LIA, ARCH, and TPPA were 100% (18/18), 100% (17/17), and 100% (18/18), and 100% (558/558), 99.8% (552/553), and 99.6% (556/558), respectively. In pregnant women, the specificity of LIA and ARCH was 100% (317/317) and of TPPA 99.7% (316/317). One sample from a child with assumed exposure to maternal antitreponemal antibodies was omitted from analysis. LIA, ARCH, and TPPA were also positive in all investigated sera from patients with known syphilis. Both automated CLIA demonstrated excellent diagnostic sensitivity and specificity when evaluated as a screening test for syphilis under routine conditions of a diagnostic laboratory. Thus, these may be used independently as an alternative to the manual TPPA screen.

Co-circulation of Toscana virus and Punique virus in northern Tunisia: a microneutralisation-based seroprevalence study.

Directory of Open Access Journals (Sweden)

Sonia Sakhria

Full Text Available BACKGROUND: In northern Tunisia, the co-circulation of two related sand fly-borne phleboviruses, Toscana virus (TOSV and Punique virus (PUNV was previously demonstrated. In contrast to TOSV, a prominent human pathogen, there is no data supporting that PUNV is capable to infect and cause disease to humans. We studied the respective involvement of TOSV and PUNV in human infections in northern Tunisia through a seroprevalence study. METHODS: The presence of TOSV and PUNV neutralising antibodies (NT-Ab was tested in human sera collected from 5 districts of the governorate of Bizerte, and the titres of NT-Ab were estimated by microneutralisation (MN assay. PRINCIPAL FINDINGS: A total of 1,273 sera were processed. TOSV and PUNV NT-Ab were detected in 522 (41% and 111 sera (8.72% respectively. TOSV seroprevalence varied from 17.2% to 59.4% depending on the district. Analysis of TOSV geometric mean titre values demonstrated a constant increase according to the age. The vast majority of sera containing NT-Ab were found to be more reactive toward TOSV than PUNV. Indeed, past infections with PUNV and TOSV were undisputable for 5 and 414 sera, respectively. CONCLUSIONS: PUNV may be capable to infect humans but at a low rate. TOSV is responsible for the vast majority of human infections by sand fly-borne phleboviruses in northern Tunisia. TOSV must be considered by physician and tested in diagnostic laboratories for patients with meningitis and unexplained fever in northern Tunisia.

Sero-epidemiological assessment and diagnosis of visceral leishmaniasis in an endemic locality using Fast Agglutination Screening Test (FAST)

NARCIS (Netherlands)

Hailu, A.; Kroon, C. C. M.; Schoone, G. J.; Berhe, N.; Schallig, H. D. F. H.; Kager, P. A.

2002-01-01

The Fast Agglutination Screening Test (FAST) was employed on sera obtained from an endemic area of visceral leishmaniasis in southwestern Ethiopia, in February 2000. The study involved (i) active case detection among 1575 residents of two villages; and (ii) passive case detection in an outpatient

The chronic obstructive pulmonary disease assessment test improves the predictive value of previous exacerbations for poor outcomes in COPD.

Science.gov (United States)

Miravitlles, Marc; García-Sidro, Patricia; Fernández-Nistal, Alonso; Buendía, María Jesús; Espinosa de Los Monteros, María José; Esquinas, Cristina; Molina, Jesús

2015-01-01

Chronic obstructive pulmonary disease (COPD) exacerbations have a negative impact on the quality of life of patients and the evolution of the disease. We have investigated the prognostic value of several health-related quality of life questionnaires to predict the appearance of a composite event (new ambulatory or emergency exacerbation, hospitalization, or death) over a 1-year follow-up. This was a multicenter, prospective, observational study. Patients completed four questionnaires after recovering from an exacerbation (COPD Assessment Test [CAT], a Clinical COPD Questionnaire [CCQ], COPD Severity Score [COPDSS], and Airways Questionnaire [AQ20]). Patients were followed-up until the appearance of the composite event or for 1 year, whichever came first. A total of 497 patients were included in the study. The majority of them were men (89.7%), with a mean age of 68.7 (SD 9.2) years, and a forced expiratory volume in 1 second of 47.1% (SD 17.5%). A total of 303 (61%) patients experienced a composite event. Patients with an event had worse mean scores of all questionnaires at baseline compared to patients without event: CAT=12.5 vs 11.3 (P=0.028); CCQ=2.2 vs 1.9 (P=0.013); COPDSS=12.3 vs 10.9 (P=0.001); AQ20=8.3 vs 7.5 (P=0.048). In the multivariate analysis, only previous history of exacerbations and CAT score ≥13.5 were significant risk factors for the composite event. A CAT score ≥13.5 increased the predictive value of previous exacerbations with an area under the receiver operating characteristic curve of 0.864 (95% CI: 0.829-0.899; P=0.001). The predictive value of previous exacerbations significantly increased only in one of the four trialled questionnaires, namely in the CAT questionnaire. However, previous history of exacerbations was the strongest predictor of the composite event.

Development and evaluation of diagnostic tests for the serological diagnosis of brucellosis in swine

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Tiziana Di Febo

2012-06-01

Full Text Available A competitive enzyme-linked immunosorbent assay (c-ELISA, an indirect ELISA (i-ELISA and a dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA were developed to test for antibodies to Brucella suis in pig and wild boar sera. An anti-Brucella-LPS monoclonal antibody (MAb 4B5A (c-ELISA and DELFIA and an anti-swine IgG monoclonal antibody (MAb 10C2G5 (i-ELISA were used for the three assays. The specificity (Sp and sensitivity (Se of the assays gave the following results: Se and Sp = 100% at a cut-off value of 61.0% (B/B0% for c-ELISA; Sp = 99.1% and Se = 100% at a cut-off value of 21.7% (percentage positivity: PP% for i-ELISA; Sp = 91.0% and Se = 75% at a cut-off value of 37.0% (B/B0% for DELFIA. In addition, the performance of a commercial fluorescence polarisation assay (FPA, standardised for bovine sera, was evaluated in swine sera. The specificity and sensitivity obtained were both 100% at a cut-off value of 99.5 (millipolarisation unit values. These results suggest that the combination of c-ELISA, i-ELISA and FPA can be used to improve the serological diagnosis of swine brucellosis.

12th International Conference on Software Engineering, Arti fi cial Intelligence Research, Management and Applications (SERA 2014)

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Software Engineering Research, Management and Applications

2015-01-01

This edited book presents scientific results of the 12th International Conference on Software Engineering, Artificial Intelligence Research, Management and Applications (SERA 2014) held on August 31 – September 4, 2014 in Kitakyushu, Japan.  The aim of this conference was to bring together researchers and scientists, businessmen and entrepreneurs, teachers, engineers, computer users, and students to discuss the numerous fields of computer science and to share their experiences and exchange new ideas and information in a meaningful way. Research results about all aspects (theory, applications and tools) of computer and information science, and to discuss the practical challenges encountered along the way and the solutions adopted to solve them. This publication captures 17 of the conference’s most promising papers.

Allergy to fish collagen: Thermostability of collagen and IgE reactivity of patients' sera with extracts of 11 species of bony and cartilaginous fish

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Yukihiro Kobayashi

2016-10-01

Conclusions: The IgE reactivity of patients' sera to fish collagen in extracts was retained even when fish meat was treated by a high heating load. As for the fish collagen, the IgE reactivities to cartilaginous fish were lower than that to bony fish. Reducing IgE reactivity to fish meat using heat is difficult, and other modalities will be required to produce hypoallergenic fish meat.

A comparison of the absorbed fluorescent treponemal antibody (FTA-ABS) test and other screening tests for treponemal disease in patients attending a venereal disease clinic.

Science.gov (United States)

Wilkinson, A E; Scrimgeour, G; Rodin, P

1972-05-01

Screening tests-absorbed fluorescent treponemal (FTA-ABS), the Reiter protein complement-fixation (RPCFT), VDRL slide test, automated reagin-and cardiolipin Wassermann reaction-were carried out on 1922 consecutive new patients attending the Whitechapel Clinic over a three-month period.Taking the FTA-ABS test results as an index, the most efficient combination of conventional tests was found to be the RPCFT and automated reagin test. The cardiolipin WR proved to be under-sensitive and of little value compared with the other tests.Forty-two per cent of the 107 sera reactive in the FTA-ABS test were not detected by the RPCFT or ART tests. An assessment based on the TPI test results and clinical findings in these patients is presented. The scope and limitations of the FTA-ABS test as a screening procedure are discussed.

The cytotoxic effect of neonatal lupus erythematosus and maternal sera on keratinocyte cultures is complement-dependent and can be augmented by ultraviolet irradiation

Energy Technology Data Exchange (ETDEWEB)

Yu, H.-S.; Chang, C.-H.; Kang, J.-W. [Kaohsiung Medical College (Taiwan). Dept. of Dermatology; Chiang, L.-C. [Kaohsiung Medical College (Taiwan). Dept. of Microbiology and Immunology; Yu, C.-L. [National Yang-Ming University School of Medicine (Taiwan). Veterans General Hospital-Taipei

1996-08-01

To elucidate the role of autoantibodies and ultraviolet (UV) exposure in the pathogenesis of the skin lesions in neonatal lupus erythematosus (NLE), keratinocytes were cultured, as the target cells, from a patient with NLE and from a normal neonate. We demonstrated that the expression of nuclear/cytoplasma Ro/SSA and La/SSB molecules on to the surface of NLE keratinocytes occurred to a much greater extent than that on normal keratinocytes. A dose of 200 mJ/cm{sup 2} UVB irradiation on NLE keratinocytes induced a 2.5-3-fold increase in Ro/SSA and La/SSB expression compared to non-irradiated cells. Sera derived from both the NLE patient and from his mother exhibited a cytotoxic effect on NLE keratinocytes, but not on control cells, in the presence of complement. Furthermore, the cytotoxicity of the sera was enhanced in UVB-irradiated NLE keratinocytes, whereas it had no cytotoxic effects on UVB-irradiated control cells. This suggests that the abnormal expression of both Ro/SSA and La/SSB on the surface membrane of NLE keratinocytes induces the autoantibodies and complements to injure the cells. This complement-mediated cytotoxic effect can be augmented by UV irradiation, a concept not incompatible with the exacerbation of the skin eruption in sun-exposed skin sites. (author).

The cytotoxic effect of neonatal lupus erythematosus and maternal sera on keratinocyte cultures is complement-dependent and can be augmented by ultraviolet irradiation

International Nuclear Information System (INIS)

Yu, H.-S.; Chang, C.-H.; Kang, J.-W.; Chiang, L.-C.; Yu, C.-L.

1996-01-01

To elucidate the role of autoantibodies and ultraviolet (UV) exposure in the pathogenesis of the skin lesions in neonatal lupus erythematosus (NLE), keratinocytes were cultured, as the target cells, from a patient with NLE and from a normal neonate. We demonstrated that the expression of nuclear/cytoplasma Ro/SSA and La/SSB molecules on to the surface of NLE keratinocytes occurred to a much greater extent than that on normal keratinocytes. A dose of 200 mJ/cm 2 UVB irradiation on NLE keratinocytes induced a 2.5-3-fold increase in Ro/SSA and La/SSB expression compared to non-irradiated cells. Sera derived from both the NLE patient and from his mother exhibited a cytotoxic effect on NLE keratinocytes, but not on control cells, in the presence of complement. Furthermore, the cytotoxicity of the sera was enhanced in UVB-irradiated NLE keratinocytes, whereas it had no cytotoxic effects on UVB-irradiated control cells. This suggests that the abnormal expression of both Ro/SSA and La/SSB on the surface membrane of NLE keratinocytes induces the autoantibodies and complements to injure the cells. This complement-mediated cytotoxic effect can be augmented by UV irradiation, a concept not incompatible with the exacerbation of the skin eruption in sun-exposed skin sites. (author)

Aspectos da padronização de testes sorológicos para a doença de Chagas: um teste imunoenzimático para a triagem de doadores de sangue Standardization of serological tests in Chagas' disease: immunoenzimatic test for screening of blood bank donnors

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A. Walter Ferreira

1991-04-01

reagent (bioELISA cruzi, Biolab Diagnóstica S/A, Brasil, was tested in serum from 1648 patients 219 with Chagas disease and 104 with other diseases, plus a comparison with well standardized immunofluorescence and hemagglutination tests in 1325 sera. In the immunoenzimatic assays, the cut off was indicated by the absorbance value of a chagasic serum showing a minimal reactivity. ELISA sensibility was 0,9954 and specificity 0,9969, as conegativity. False positive results were absent with sera from sifilis, toxoplasmosis, mononucleosis and high titered sera for antistreptolysin 0 antibodies. However they were seen in 5 to 15 cases of tegu-mentar leishmaniosis, 1 of 12 Kala-azar 1 of 15 reumatoid arthritis and 1 of 12 sistemic lupus erythematosus. The high sensibility in chagasics and high specificity in the general population indicate the confiability of the imunoenzimatic assay for screening blood donors and even to confirm a clinical diagnosis of Chagas' disease.

Antibodies to adult T-cell leukemia-virus-associated antigen (ATLA) in sera from patients with ATL and controls in Japan: a nation-wide sero-epidemiologic study.

Science.gov (United States)

Hinuma, Y; Komoda, H; Chosa, T; Kondo, T; Kohakura, M; Takenaka, T; Kikuchi, M; Ichimaru, M; Yunoki, K; Sato, I; Matsuo, R; Takiuchi, Y; Uchino, H; Hanaoka, M

1982-06-15

A nation-wide sero-epidemiologic survey of adult T-cell leukemia virus (ATLV), detected es anti-ATLA (ATLV-associated antigen), was made in Japan. Sera from adult donors in 15 different locations were screened for anti-ATLA. High incidences (6 to 37%) of antibody-positive donors were found in seven regions, one in northern Japan, and the others in southwestern regions. These areas are ATLV-endemic areas corresponding to ATL-endemic areas. Examination of sera from healthy donors aged 6 to 80 years in ATL-endemic areas showed an age-dependent increase of seropositive donors with a maximum of about 30% at 40 years of age. Anti-ATLA was found in all but two of 142 patients with ATL. Anti-ATLA-positive patients with ATL were mainly found in ATLV-endemic areas, and only a few in ATL-nonendemic areas. Six patients with cutaneous T-cell lymphoma in ATLV-nonendemic areas gave a negative reaction for anti-ATLA. The geometric mean titer of anti-ATLA of patients with ATL was higher than that of healthy donors.

SEROLOGICAL DIAGNOSIS OF MODERN CHOLERA USING LIPOSOMAL ENTEROTOXIC DIAGNOSTICUM IN COMPLEMENT FIXATION TEST

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I. V. Savelyeva

2013-01-01

Full Text Available Abstract. The possibility of serological diagnosis of cholera using cholera enterotoxic diagnostics kit in complement fixation test to detect anti-enterotoxic antibodies in sera of patients with cholera caused by hybrid variants of the El Tor biovar has been demonstrated. In patients with mild course of cholera anti-enterotoxic antibodies were detected in titres 1:50 and 1:200 in paired sera obtained on the 7th and 14th days of disease, respectively (fourfold titre increase. In patients with the course of medium severity 32-fold titre increase was recorded from the titre 1:100 in serum obtained on the fifth day of disease till the titre 1:3200 — on the twelfth day of disease. Antibodies titers reached 1:1600 and 1:800 were revealed in two medium course patients (adult and infant of 10 months on the sixth day of disease.

One-shot flow injection spectrophotometric simultaneous determination of copper, iron and zinc in patients' sera with newly developed multi-compartment flow cell

International Nuclear Information System (INIS)

Teshima, Norio; Gotoh, Shingo; Ida, Kazunori; Sakai, Tadao

2006-01-01

We propose here an affordable flow injection method for simultaneous spectrophotometric determination of copper, iron and zinc in patients' sera. The use of a newly designed multi-compartment flow cell allowed the simultaneous determination of the three metals with a single injection ('one-shot') and a double beam spectrophotometer. The chemistry relied on the reactions of these metals with 2-(5-nitro-2-pyridylazo)-5-[N-propyl-N-(3-sulfopropyl)amino]phenol (nitro-PAPS) to form corresponding colored complexes. At pH 3.8, only copper-nitro-PAPS complex was formed in the presence of pyrophosphate as a masking agent for iron, and then the copper and iron(II) complexes were formed in the presence of reductant (ascorbic acid) at the same pH, and finally all three metals reacted with nitro-PAPS at pH 8.6. The characteristics were introduced into the flow system to determine each metal selectively and sensitively. Under the optimum conditions, linear calibration curves for the three metals were obtained in the range of 0.01-1 mg L -1 with a sample throughput rate of 20 h -1 . The limits of detection (3σ) were 3.9 μg L -1 for copper, 4.1 μg L -1 for iron and 4.0 μg L -1 for zinc. The proposed method was applied to analysis of some patients' sera

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

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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.

Comparison of enzyme immunoassays for the diagnosis of bovine brucellosis

International Nuclear Information System (INIS)

Nielsen, K.; Kelly, L.; Gall, D.; Balsevicius, S.; Bosse, J.; Kelly, W.; Nicoletti, P.

1998-01-01

The indirect enzyme immunoassay for measurement of bovine antibody to Brucella abortus was tested on 15,716 Canadian sera to assess the specificity. These sera were also tested by the buffered plate antigen test. Two ELISA formats were used for assessment of data: the targeting procedure using a positive control serum allowed to develop to an optical density of 1.0 and the use of a positive control serum to determine relative positivity at a set time. Two different cut-off values were also assessed for each assay. A total of 763 sera gave reactions above established cut-off values in the ELISA while 216 were positive in the buffered plate antigen test (BPAT). A modification of the indirect ELISA employed divalent cation chelating agents (EDTA/EGTA) incorporated into the serum incubation stage to eliminate some non-specific reactions. This method was applied only to the 763 indirect ELISA reactor sera and it eliminated all but 93 or 37, depending on the cut-off selected, of the reactions. Sensitivity was assessed by testing 424 sera from Brucella abortus culture positive cattle. The indirect ELISA classified all 424 sera as positive by either method of data handling and with or without addition of EDTA/EGTA for a specificity estimate of 100%. In the BPAT, 412 sera gave a positive agglutination reaction. Ten percent of the 15,716 sera were randomly selected and tested by two different competitive ELISAs and by the complement fixation test (CFT). One competitive ELISA used Brucella abortus O-polysaccharide as the antigen and an enzyme conjugated monoclonal antibody to the O-polysaccharide for competition and detection. Of the sera tested, 34 gave false positive reactions. On a retest, the false positive reactions were reduced to 2. The second competitive ELISA used lipopolysaccharide as the antigen, a different monoclonal antibody but also specific for the O-polysaccharide for competition and commercially available goat anti-mouse IgG enzyme conjugate for detection

Human CRISP-3 binds serum alpha(1)B-glycoprotein across species

DEFF Research Database (Denmark)

Udby, Lene; Johnsen, Anders H; Borregaard, Niels

2010-01-01

CRISP-3 was previously shown to be bound to alpha(1)B-glycoprotein (A1BG) in human serum/plasma. All mammalian sera are supposed to contain A1BG, although its presence in rodent sera is not well-documented. Since animal sera are often used to supplement buffers in experiments, in particular...

Validation of a foot-and-mouth disease antibody ELISA in five Latin American countries

International Nuclear Information System (INIS)

Sondahl, M.S.; Penha Dias Gomes, M. da; Aurnheimer Martins, M.; Washington Lopez, J.

1998-01-01

The work plan consisted of using a liquid phase blocking ELISA test for the detection of antibodies to foot-and-mouth disease virus (FMDV) using the following categories of sera: (A) Spot test 120 non-infected/non-vaccinated bovine sera diluted 1:32; (B) Titration test: 120 bovine sera from animals vaccinated with trivalent oil vaccine, bled 30 days after vaccination; (C) Titration test with sera from non-infected/non-vaccinated bovines that presented titers >1:32 in the spot test. To detect FMD positive animals in the field, the spot test established with a cut-off of 1: 32 demonstrated in this work a good specificity with the non-vaccinated group, where 3 animals out of 120 were considered positive. The antibody titration test is an excellent tool to determine the level of antibodies in cattle populations. The protocol indicates that positive sera from the spot test should be tested in the titration assay in a starting dilution of 1:32. We suggest to use a lower starting dilution (1:16) in order to start below the discriminative of positive spot test sera 1:32 for the titration assay procedures. (author)

Has introduction of rapid drug susceptibility testing at diagnosis impacted treatment outcomes among previously treated tuberculosis patients in Gujarat, India?

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Paresh Dave

Full Text Available Revised National TB Control Programme (RNTCP in India recommends that all previously-treated TB (PT patients are offered drug susceptibility testing (DST at diagnosis, using rapid diagnostics and screened out for rifampicin resistance before being treated with standardized, eight-month, retreatment regimen. This is intended to improve the early diagnosis of rifampicin resistance and its appropriate management and improve the treatment outcomes among the rest of the patients. In this state-wide study from Gujarat, India, we assess proportion of PT patients underwent rapid DST at diagnosis and the impact of this intervention on their treatment outcomes.This is a retrospective cohort study involving review of electronic patient-records maintained routinely under RNTCP. All PT patients registered for treatment in Gujarat during January-June 2013 were included. Information on DST and treatment outcomes were extracted from 'presumptive DR-TB patient register' and TB treatment register respectively. We performed a multivariate analysis to assess if getting tested is independently associated with unfavourable outcomes (death, loss-to-follow-up, failure, transfer out.Of 5,829 PT patients, 5306(91% were tested for drug susceptibility with rapid diagnostics. Overall, 71% (4,113 TB patients were successfully treated - 72% among tested versus 60% among non-tested. Patients who did not get tested at diagnosis had a 34% higher risk of unsuccessful outcomes as compared to those who got tested (aRR - 1.34; 95% CI 1.20-1.50 after adjusting for age, sex, HIV status and type of TB. Unfavourable outcomes (particularly failure and switched to category IV were higher among INH-resistant patients (39% as compared to INH-sensitive (29%.Offering DST at diagnosis improved the treatment outcomes among PT patients. However, even among tested, treatment outcomes remained suboptimal and were related to INH resistance and high loss-to-follow-up. These need to be addressed

Clinical comparison of the Treponema pallidum CAPTIA syphilis-G enzyme immunoassay with the fluorescent treponemal antibody absorption immunoglobulin G assay for syphilis testing.

Science.gov (United States)

Halling, V W; Jones, M F; Bestrom, J E; Wold, A D; Rosenblatt, J E; Smith, T F; Cockerill, F R

1999-10-01

Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphilis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results of rapid plasmid reagin (RPR) testing of the same specimens. Borderline CAPTIA-positive samples (antibody indices of >/=0.650 and 0.900, the sample was considered positive. Thirteen of 89 (15%) samples had discrepant results. Compared to the FTA-ABS assay, the CAPTIA EIA had a sensitivity and specificity and positive and negative predictive values of 70.7, 97.9, 96.7, and 79.7%, respectively. In another analysis, discrepancies between results were resolved by repeated FTA-ABS testing (technologists were blinded to previous RPR results) and patient chart reviews. Seven CAPTIA-negative samples which were previously interpreted (unblinded) as minimally reactive by the FTA method were subsequently interpreted (blinded) as nonreactive. One other discrepant sample (CAPTIA negative and FTA-ABS positive [at an intensity of 3+], unblinded) was FTA negative with repeated testing (blinded). For the five remaining discrepant samples, chart reviews indicated that one patient (CAPTIA negative and FTA-ABS positive [minimally reactive], blinded) had possible syphilis. These five samples were also evaluated and found to be negative by another treponema-specific test, the Treponema pallidum microhemagglutination assay. Therefore, after repeated testing and chart reviews, 2 of the 89 (2%) samples had discrepant results; the adjusted sensitivity, specificity, and positive and negative predictive values were 96.7, 98.3, 96.7, and 98.3%, respectively. This study demonstrates that the CAPTIA IgG EIA is a reliable method for syphilis testing and that personnel performing tests

Bioactivity and immunoactivity of growth hormone during dynamic testing of patients with acromegaly

International Nuclear Information System (INIS)

Baldwin, A.L.; Norman, M.; Butler, J.; Aston, R.; Buchanan, C.

1988-01-01

We have used the Nb2 cell proliferation bioassay for lactogenic hormones to investigate the biological activity of hGH in sera of patients with acromegaly. The specificity of the assay has been improved by the use of monoclonal antibodies to block the activity of individual lactogenic hormones. Disease activity in patients was assessed by scoring signs and symptoms, and by measuring IGF-I concentrations in some patients. Patients with a wide spectrum of disease activity were studied using a TRH test. hGH concentrations were measured by bioassay, RIA and immunoradiometric assay (IRMA) at 0,20 and 60 min after injection of TRH. There was a high degree of correlation between log 10 of all hGH concentrations measured by bioassay and RIA (r = 0.995, P<0.0001), between bioassay and IRMA (r = 0.990, P<0.0001), and between RIA and IRMA (r = 0.995, P<0.0001). In contrast to previous reports, we found no evidence for changes in the bioactivity of hGH secreted after pituitary stimulation. (author)

Development of an enzyme-linked immunoelectrotransfer blot (EITB) assay using two baculovirus expressed recombinant antigens for diagnosis of Taenia solium taeniasis.

Science.gov (United States)

Levine, Min Z; Lewis, Melissa M; Rodriquez, Silvia; Jimenez, Juan A; Khan, Azra; Lin, Sehching; Garcia, Hector H; Gonzales, Armando E; Gilman, Robert H; Tsang, Victor C W

2007-04-01

Taeniasis diagnosis is an important step in the control and elimination of both cysticercosis and taeniasis. We report the development of 2 serological taeniasis diagnostic tests using recombinant antigens rES33 and rES38 expressed by baculovirus in insect cells in an EITB format. In laboratory testing with defined sera from nonendemic areas, rES33 has a sensitivity of 98% (n = 167) and a specificity of 99% (n = 310) (J index: 0.97); rES38 has a sensitivity of 99% (n = 146) and a specificity of 97% (n = 275) (J index: 0.96). Independent field testing in Peru showed 97% (n = 203) of the taeniasis sera were positive with rES33, and 100% of the nontaeniasis sera (n = 272) were negative with rES33; 98% (n = 198) of taeniasis sera were positive with rES38, and 91% (n = 274) of the nontaeniasis sera were negative with rES38. Among the Peruvian sera tested, 17 of 26 Peruvian Taenia saginata sera were false positive with rES38 test. Both tests were also examined with cysticercosis sera, with a positive rate ranging from 21% to 46%. rES33 and rES38 tests offer sensitive and specific diagnosis of taeniasis and easy sample collection through finger sticks that can be used in large-scale studies. They are currently being used in cysticercosis elimination programs in Peru.

Correlation analysis of trace elemental data obtained from blood sera of ovarian cancer patients using PIXE

International Nuclear Information System (INIS)

Naidu, B.G.; Sarita, P.; Naga Raju, G.J.

2017-01-01

Proton induced X-ray emission (PIXE) technique is used for analysis of trace elements present in the blood sera of ovarian cancer patients and healthy controls. This work is also intended to establish the role played by trace elements in carcinogenic process. It is observed that the concentrations of elements Ti, V, Cr, Mn, Fe, Ni, Rb and Sr are lower and the concentration of Cu is higher in the cancer patients when compared to controls. However, no change in concentration is found in the elements Co, Zn, As, Se and Br. Correlation analysis of the data using SPSS 16.0 has revealed a strong positive correlation between Ti-V, Ni-Co, Cu-Fe, As-Ti, Br-Ti, Br-V and Sr-Fe while strong negative correlations are observed for Cu-Ti, As-Cu and Br-Cu. Changes in trace elemental content are probably associated with ovarian carcinogenesis. (author)

Low maternal vitamin D as a risk factor for schizophrenia: a pilot study using banked sera.

Science.gov (United States)

McGrath, John; Eyles, Darryl; Mowry, Bryan; Yolken, Robert; Buka, Stephen

2003-09-01

Evidence from epidemiology suggests that low maternal vitamin D may be a risk factor for schizophrenia. Based on sera taken during the third trimester, we compared the level of 25 hydroxyvitamin D3 in mothers of individuals with schizophrenia or schizoaffective disorders versus mothers of unaffected controls. For each case, we selected two controls matched on race, gender and date of birth of the offspring. There was no significant difference in third trimester maternal vitamin D in the entire sample (cases = 26, controls = 51). Within the subgroup of black individuals (n = 21), there was a trend level difference in the predicted direction. Maternal vitamin D does not operate as a continuous graded risk factor for schizophrenia, however, the results in the black subgroup raise the possibility that below a certain critical threshold, low levels of maternal vitamin D may be associated with an increased risk of schizophrenia.

Transmission of Hepatitis C Virus From Organ Donors Despite Nucleic Acid Test Screening.

Science.gov (United States)

Suryaprasad, A; Basavaraju, S V; Hocevar, S N; Theodoropoulos, N; Zuckerman, R A; Hayden, T; Forbi, J C; Pegues, D; Levine, M; Martin, S I; Kuehnert, M J; Blumberg, E A

2015-07-01

Nucleic acid testing (NAT) for hepatitis C virus (HCV) is recommended for screening of organ donors, yet not all donor infections may be detected. We describe three US clusters of HCV transmission from donors at increased risk for HCV infection. Donor's and recipients' medical records were reviewed. Newly infected recipients were interviewed. Donor-derived HCV infection was considered when infection was newly detected after transplantation in recipients of organs from increased risk donors. Stored donor sera and tissue samples were tested for HCV RNA with high-sensitivity quantitative PCR. Posttransplant and pretransplant recipient sera were tested for HCV RNA. Quasispecies analysis of hypervariable region-1 was used to establish genetic relatedness of recipient HCV variants. Each donor had evidence of injection drug use preceding death. Of 12 recipients, 8 were HCV-infected-6 were newly diagnosed posttransplant. HCV RNA was retrospectively detected in stored samples from donor immunologic tissue collected at organ procurement. Phylogenetic analysis showed two clusters of closely related HCV variants from recipients. These investigations identified the first known HCV transmissions from increased risk organ donors with negative NAT screening, indicating very recent donor infection. Recipient informed consent and posttransplant screening for blood-borne pathogens are essential when considering increased risk donors. © Copyright 2015 The American Society of Transplantation and the American Society of Transplant Surgeons.

Evaluation of coproexamination as a diagnostic test for avian botulism

Science.gov (United States)

Jensen, Wayne I.

1981-01-01

Fecal extracts and blood sera from 113 ducks showing clinical signs of botulism were examined for Clostridium botulinum type C toxin by means of the mouse toxicity test to evaluate coproexamination as a diagnostic procedure, as compared with demonstration of toxin in serum. When death of test mice unprotected with type specific antitoxin (while protected controls survived) was the criterion, 78.8% of the sera and 5.3% of the fecal extracts were positive. When characteristic signs of intoxication in the unprotected mice was included as evidence of toxin in the specimens, these percentages increased to 86.7 and 6.2, respectively.Fecal specimens were collected hourly for the first 6 h after peroral dosing of eight mallards (Anas platyrhynchos) with 1.0 LD50, of type C toxin and at 24, 48, and 72 h from birds surviving that long. From 2 to 4 toxin-positive specimens were passed by all eight ducks during the first 6 h, five specimens were positive at 24 h, and three were positive at 48 h. Only three specimens were collected at 72 h, all of which were negative. These findings suggest that attempts to detect toxin in the feces of wild ducks might have been more successful had the birds been captured earlier in the course of the disease.

Immunodiagnosis of Trypanosoma cruzi (Chagas' Disease Infection in Naturally Infected Dogs

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Lauricella MA

1998-01-01

Full Text Available This study reports on the standardization of an enzyme-linked immunosorbent assay (ELISA for detecting specific antibodies anti-Trypanosoma cruzi in naturally infected dogs. Sera from 182 mongrel dogs of all ages residing in four rural villages in Santiago del Estero, Argentina, were collected in November 1994 and preserved in buffered neutral glycerin. All sera were tested by indirect hemagglutination test (IHAT, indirect immunofluorescence test (IFAT, and ELISA using the flagellar fraction of T. cruzi as antigen. Dog sera from an area without vectorial transmission were used to calculate ELISA specificity and cut-off value. Eighty-six percent of sera had concordant results for all tests. All sera reactive for IHAT and IFAT were also reactive for ELISA, except in one case. Sera tested by ELISA when diluted 1:200 allowed a clearer division between non-reactive and reactive sera than when 1:100 with greater agreement among serologic techniques. The specificity of ELISA was 96.2%. Among 34 adult dogs with a positive xenodiagnosis, sensitivity was 94% both for ELISA and IFAT. ELISA is the first choice for screening purposes and one of the pair of techniques recommended for diagnostic studies in dog populations

Comparison of an enzyme linked immunosorbent assay (ELISA) and a radioallergosorbent test (RAST) for detection of IgE antibodies to Brugia malayi

NARCIS (Netherlands)

Wahyuni, Sitti; van Ree, Ronald; Mangali, Andarias; Supali, Taniawati; Yazdanbakhsh, Maria; Sartono, Erliyani

2003-01-01

The enzyme linked immunosorbent assay (ELISA) for specific IgE antibodies to Brugia malayi was compared with the radioallergosorbent test (RAST) for use in immunoepidemiological studies of lymphatic filariasis. Sera used were from individuals (aged 5-82 years) living in an area endemic for lymphatic

Evaluation of enzyme-linked immunosorbent and alternative assays for detection of HIV antibodies using panels of Brasilian sera Avaliação de testes sorológicos para a detecção de anticorpos anti-HIV em painéis de soros de brasileiros

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Jairo Ivo-Dos-Santos

1990-04-01

Full Text Available Sera from 472 Brazilian subjects, confirmed to be either positive or negative for HIV antibodies and comprising the total clinical spectrum of HIV infection, were utilized in the evaluation of six commercially available enzyme-linked immunosorbent assays (ELISA, as well as of four alternative assays, namely indirect immunofluorescence (IIF, passive hemagglutination (PHA, dot blot and Karpas AIDS cell test. The sensitivities ranged from 100% (Abbott and Roche ELISA to 84.2% (PHA and the specificities ranged from 99.3% (IIF to 80.2% (PHA. The sensitivity and specificity of the PHA and the sensitivity of the Karpas cell test were significantly lower than those of the other tests. Although the IFF and dot blot had good sensitivities and specificities, the six ELISA were more attractive than those tests when other parameters such as ease of reading and duration of assay were considered.Os soros de 472 brasileiros, confirmados como sendo positivos ou negativos em relação à presença de anticorpos anti-HIV e compreendendo todo o espectro clínico da infecção, foram utilizados na avaliação de seis ensaios imunoenzimáticos comerciais (ELISA, bem como de quatro testes alternativos tais como imunofluorescência indireta (IFI, hemaglutinação passiva (HP, dot blot e Karpas AIDS cell test. As sensibilidades variaram de 100% (ELISA Abbott e Roche a 84,2% (HP e as especificidades variaram de 99,3% (IFI a 80,2% (HP. A sensibilidade e especificidade da HP e a sensibilidade do Karpas AIDS cell test foram significativamente menores que os outros ensaios. Embora a IFI e o dot blot tivessem apresentado uma boa sensibilidade e especificidade, os ensaios imunoenzimáticos (ELISA foram mais adequados para serem utilizados em triagem quando outros parâmetros tais como facilidade de leitura e interpretação dos resultados e duração dos ensaios foram considerados.

A Cross-Sectional Survey of HIV Testing and Prevalence in Twelve Brazilian Correctional Facilities.

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Renata Viebrantz Enne Sgarbi

Full Text Available Prior studies have reported higher HIV prevalence among prisoners than the general population in Brazil, but data have been derived from single prisons. The aim of this study was to evaluate HIV testing practices, prevalence and linkage to care among inmates in a network of 12 prisons.We administered a questionnaire to a population-based sample of inmates from 12 prisons in Central-West Brazil and collected sera for HIV and syphilis testing from January to December 2013. We evaluated factors associated with HIV testing and infection using multivariable logistic regression models. Six months after HIV testing, we assessed whether each HIV-infected prisoner was engaged in clinical care and whether they had started antiretroviral therapy.We recruited 3,362 inmates, of whom 2,843 (85% were men from 8 prisons, and 519 (15% were women from 4 prisons. Forty-five percent of participants reported never having been tested for HIV previously. In multivariable analysis, the variables associated with previous HIV testing were lack of a stable partner (adjusted odds ratio [AOR]: 1.38; 95% CI: 1.18-1.60, completed more than four years of schooling (AOR 1.40; 95% CI: 1.20-1.64, history of previous incarceration (AOR: 1.68; 95% CI: 1.43-1.98, history of mental illness (AOR 1.52; 95% CI: 1.31-1.78 and previous surgery (AOR 1.31; 95% CI: 1.12-1.52. Fifty-four (1.6% of all participants tested positive for HIV; this included 44 (1.54% men and 10 (1.92% women. Among male inmates, HIV infection was associated with homosexuality (AOR 6.20, 95% CI: 1.73-22.22, self-report of mental illness (AOR 2.18, 95% CI: 1.13-4.18, history of sexually transmitted infections (AOR 3.28, 95% CI: 1.64-6.56, and syphilis sero-positivity (AOR 2.54, 95% CI: 1.20-5.39. Among HIV-infected individuals, 34 (63% were unaware of their HIV status; only 23 of these 34 (68% newly diagnosed participants could be reached at six month follow-up, and 21 of 23 (91% were engaged in HIV care.HIV testing

Comparison between the Counter Immunoelectrophoresis Test and Mouse Neutralization Test for the Detection of Antibodies against Rabies Virus in Dog Sera

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Luzia Helena Queiroz da Silva

2002-03-01

Full Text Available The detection of rabies antibodies is extremely valuable for epidemiological studies, determination of immune status in man, animals, and for the diagnosis of the disease. Several serological procedures have been described for this purpose. The present study reports a comparison between counterimmunoelectrophoresis test (CIET and mouse neutralization test (MNT in the detection of antibodies against rabies virus from 212 serum samples of vaccinated dogs. The agreement between both techniques was 79.7% and a significative association was demonstrated. The correlation coefficients between MNT and the CIET titers was determined considering 88 samples showing positive results in both techniques [CIET = 2 and MNT = 5 (0.13 IU/ml] and resulted r² = 0.7926 (p < 0.001. The performance of CIET system was technically simple, cheap and rapid, and thereby it could be useful for serological monitoring of dog vaccination campaigns as well as for individual analysis.

Seroprevalence of human brucellosis in and around Jammu, India, using different serological tests

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H. K. Sharma

2016-07-01

Full Text Available Aim: Brucellosis is a disease of zoonotic importance as it affects both human as well as animal’s health, and therefore, directly affects animal productivity and human efficiency. Therefore, a study was conducted to estimate the seroprevalence of brucellosis in humans in Jammu and surrounding areas. Materials and Methods: A total of 121 sera samples from humans occupied with professional related to animals were collected and tested for anti-Brucella antibodies by Rose Bengal plate test (RBPT, modified RBPT (mRBPT, standard tube agglutination test (STAT, and indirect enzyme-linked immunosorbent assay (I-ELISA. Sampling was done keeping in view with the occupation, sex, and age. Results: The overall seroprevalence of brucellosis recorded was 4.96%. The test-wise seroprevalence was 9.91% by RBPT, 9.91% by mRBPT, 9.09% by STAT, and 16.52% by I-ELISA. The prevalence of brucellosis was higher in >35-50 years age group compared to >20-35 years and >50-65 years. Sex-wise seroprevalence was higher in males than females. Taking I-ELISA as standard, the relative sensitivities of mRBPT, RBPT, and I-ELISA were in the order of mRBPT=RBPT>STAT. All the tests revealed high specificity values; however, among different serological tests, I-ELISA detected a maximum number of positive sera samples. Conclusions: The prevalence of brucellosis was found to be approximately 5%. The adult (>35-50 years age male group was most vulnerable. The routine diagnosis of brucellosis involved the conventional serological tests, viz., RBPT and STAT, but each was associated with drawbacks which could give either false-positive or false-negative interpretation. Therefore, it is always recommended to use a battery of tests in the diagnosis of brucellosis.

Epitope-blocking enzyme-linked immunosorbent assay to differentiate west nile virus from Japanese encephalitis virus infections in equine sera.

Science.gov (United States)

Kitai, Yoko; Shoda, Mizue; Kondo, Takashi; Konishi, Eiji

2007-08-01

West Nile virus (WNV) is now widely distributed worldwide, except in most areas of Asia where Japanese encephalitis virus (JEV) is distributed. Considering the movement and migration of reservoir birds, there is concern that WNV may be introduced in Asian countries. Although manuals and guidelines for serological tests have been created in Japan in preparedness for the introduction of WNV, differential diagnosis between WNV and JEV may be complicated by antigenic cross-reactivities between these flaviviruses. Here, we generated a monoclonal antibody specific for the nonstructural protein 1 (NS1) of WNV and established an epitope-blocking enzyme-linked immunosorbent assay that can differentiate WNV from JEV infections in horse sera. Under conditions well suited for our assay system, samples collected from 95 horses in Japan (regarded as negative for WNV antibodies), including those collected from horses naturally infected with JEV, showed a mean inhibition value of 8.2% and a standard deviation (SD) of 6.5%. However, inhibition values obtained with serum used as a positive control (obtained after 28 days from a horse experimentally infected with WNV) in nine separate experiments showed a mean of 54.4% and an SD of 7.1%. We tentatively determined 27.6% (mean + 3 x SD obtained with 95 negative samples) as the cutoff value to differentiate positive from negative samples. Under this criterion, two horses experimentally infected with WNV were diagnosed as positive at 12 and 14 days, respectively, after infection.

Multiple free-radical scavenging (MULTIS) capacity in cattle serum.

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Sueishi, Yoshimi; Kamogawa, Erisa; Kimura, Anna; Kitahara, Go; Satoh, Hiroyuki; Asanuma, Taketoshi; Oowada, Shigeru

2017-01-01

Multiple free-radical scavenging (MULTIS) activity in cattle and human sera was evaluated with electron spin resonance spectroscopy. Scavenging rates against six active species, namely hydroxyl radical, superoxide anion, alkoxyl radical, alkylperoxyl radical, methyl radical, and singlet oxygen were quantified. The difference in the electron spin resonance signal intensity in the presence and absence of the serum was converted into the scavenging rates. Comparative MULTIS measurements were made in sera from eight beef cattle, three fetal calves and fifteen healthy human volunteers. Further, we determined the MULTIS value of albumin, the most abundant component in serum. MULTIS values in cattle sera indicated higher scavenging activity against most free radical species tested than human sera. In particular, cattle serum scavenging activities against superoxide and methyl radical were higher than human serum by 2.6 and 3.7 fold, respectively. In cattle serum, albumin appears to play a dominant role in MULTIS activity, but in human serum that is not the case. Previous data indicated that the abundance of uric acid in bovine blood is nearly 80% less than humans; however, this difference does not explain the deviation in MULTIS profile.

Antibodies to actin in autoimmune haemolytic anaemia

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Ritzmann Mathias

2010-03-01

Full Text Available Abstract Background In autoimmune haemolytic anaemia (AIHA, autoreactive antibodies directed against red blood cells are up-regulated, leading to erythrocyte death. Mycoplasma suis infections in pigs induce AIHA of both the warm and cold types. The aim of this study was to identify the target autoantigens of warm autoreactive IgG antibodies. Sera from experimentally M. suis-infected pigs were screened for autoreactivity. Results Actin-reactive antibodies were found in the sera of 95% of all animals tested. The reactivity was species-specific, i.e. reactivity with porcine actin was significantly higher than with rabbit actin. Sera of animals previously immunised with the M. suis adhesion protein MSG1 showed reactivity with actin prior to infection with M. suis indicating that molecular mimicry is involved in the specific autoreactive mechanism. A potentially cross-reactive epitope was detected. Conclusions This is the first report of autoreactive anti-actin antibodies involved in the pathogenesis of autoimmune haemolytic anaemia.

Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation

Science.gov (United States)

Molins, Claudia R.; Sexton, Christopher; Young, John W.; Ashton, Laura V.; Pappert, Ryan; Beard, Charles B.

2014-01-01

Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first- and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests. PMID:25122862

Significance of isolated reactive treponemal chemiluminescence immunoassay results.

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Hunter, Michael G; Robertson, Peter W; Post, Jeffrey J

2013-05-01

Isolated reactive serum treponemal chemiluminescence immunoassay (CIA) specimens cause clinical uncertainty. Sera were screened by CIA, and reactive samples underwent reflex testing with rapid plasma reagin (RPR), Treponema pallidum particle agglutination (TPPA), and fluorescent treponemal antibody absorption (FTA Abs) assays. Samples reactive only on the CIA were deemed "isolated" reactive CIA samples. We undertook detailed review of a subset of subjects with isolated reactive CIA specimens. Of 28 261 specimens, 1171 (4.1%) were reactive on CIA, of which 133 (11.3%) had isolated CIA reactivity. Most subjects (66 of 82 [80.5%]) with isolated reactive CIA specimens were from high-prevalence populations. We found evidence of CIA, TPPA, and FTA Abs seroreversion. The median chemiluminescent signal-to-cutoff ratio was similar for isolated reactive CIA sera and sera that were reactive on either FTA Abs or TPPA assays (2.19 vs 2.32; P = .15) but lower than for sera reactive on both FTA Abs and TPPA assays (12.37; P < .001) or for sera reactive on RPR assays (25.53; P < .001). A total of 11 of 20 patients (55%) with an isolated reactive CIA specimen who underwent medical record review had previous or subsequent evidence of syphilis infection. Isolated reactive CIA specimens may represent true T. pallidum infection and may be found after seroreversion of traditional treponemal assays.

ATLANTIC DIP: simplifying the follow-up of women with previous gestational diabetes.

LENUS (Irish Health Repository)

Noctor, E

2013-11-01

Previous gestational diabetes (GDM) is associated with a significant lifetime risk of type 2 diabetes. In this study, we assessed the performance of HbA1c and fasting plasma glucose (FPG) measurements against that of 75 g oral glucose tolerance testing (OGTT) for the follow-up screening of women with previous GDM.

Determination of triiodothyronine using RIA-Mat T3 kit

International Nuclear Information System (INIS)

Hara, Masao

1975-01-01

A RIA-Mat T 3 kit was used for an in vitro measurement of triiodothyronine in serum. A resin strip was utilized to absorb free triiodothyronine after the serum was incubated with antibody against triiodothyronine. This test was easy to manipulate and the reproducibility test with same sera, dilution test, and recovery test were also satisfactory. The normal range of triiodothyronine determined with this test kit was 119 +- 40 ng/dl using 50 test sera. Ninety sera with various thyroid diseases were also determined. (auth.)

Factors influencing Haemagglutination Tests with Tetanus Antitoxin

Science.gov (United States)

Fulthorpe, A. J.

1959-01-01

The agglutinin titres of tetanus antitoxins tested with preserved sheep cells sensitized with tetanus toxoid have been found to be inversely proportional to the cell concentration used. The agglutinin titres per international unit of antitoxin were widely different among various electrophoretically separated globulin fractions of tetanus antitoxin. There were smaller differences between the agglutinating capacity of γ-globulin fractions of sera from different horses. With haemagglutination inhibition tests there was always a relative increase in test dose between the LA and LA/100 level of test. This relative increase was greatest where the sensitivity of the cell suspensions was high. Increased suspension sensitivity may be the result of treatment of cells with greater quantities of sensitizing antigen or of reduction in the concentration of sensitized cells in the suspension. A greater volume of serum was required to give a standard end point, when mixed with a test dose (LA) of toxin, if the mixtures were allowed to stand for increasing periods of time. These observations have been discussed. PMID:13653730

Application of modified shell vial culture procedure for arbovirus detection.

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Edna R Caceda

Full Text Available The isolation of arboviruses from patient's low titer sera can be difficult. Here we compared the detection efficiency of Dengue (DEN, Yellow Fever (YF, Saint Louis Encephalitis (SLE, West Nile (WN, Ilheus (ILH, Group C (GC, Oropouche (ORO, Mayaro (MAY and Venezuela Encephalitis Equine (VEE viruses using a Modified Shell Vial Culture (MSVC protocol to a Standard Cell Culture (SCC protocol. First the MSVC and SCC protocols were compared using five dilutions for each of the following stock viruses: DEN-1, DEN-2, DEN-3, DEN-4, YF, SLE, WN, ILH, GC, ORO, MAY and VEE. Next, patients' original sera from which viruses (DEN-1, DEN-2, DEN-3, YF, GC, ORO, MAY and VEE had been previously isolated were compare by the two methods using five sera dilutions. In addition, seven sera that were positive for DEN-3 by RT-PCR and negative by SCC were processed by MSVC. The MSVC protocol was consistently 1-2 logs higher virus dilution more sensitive for virus detection than the SCC protocol for all stock Flaviviruses tested (DEN-1, DEN-2, DEN-3, DEN-4, YF, SLE, WN and ILH. MSVC was equal to or one log more sensitive for virus detection than SCC for the stock Bunyaviruses (GC and ORO. For the stock Alphavirus MAY, MSVC was equally or one log more sensitive for virus detection than SCC, while for VEE SCC was equally or one log more sensitive for virus detection than MSVC. MSVC was consistently one to two sera dilutions more sensitive than SCC for the detection of Flaviviruses from patients' sera. Both methods were approximately equally sensitive for the detection of Bunyaviruses from patients' sera and equal or one dilution less sensitive for the detection of Alphaviruses from patients' sera. Additionally, MSVC detected DEN virus in five of seven DEN-3 RT-PCR positive, SCC negative patients' sera.

[Diagnosis of human brucellosis. Role of pH in the seroagglutination test and influence of pH on the agglutinating activity of IgM, IgG and IgA antibodies].

Science.gov (United States)

Rubio Vallejo, Manuel; del Pozo, José L; Del Pozo León, José Luis; Hernández-Molina, Juan Manuel; Dorronsoro Ibero, Inés; Marrodán Ciordia, Teresa; Díaz García, Ramón

2002-04-01

To evaluate the role of pH in the seroagglutination test (SAT)and Rose Bengal (RB) test, and to determine the influence of pH on the agglutinating activity of IgM, IgG and IgA antibodies. The SAT was performed at pH 7.2 or pH 5.0 in standard microtiter-type polystyrene plates using Ring Test antigen or the Brucella suspension (BRUCAPT) provided in the Brucellacapt kits. Specific antibodies against native hapten were determined by radial immunodiffusion. Additionally, IgG, IgA and IgM fractions were separated from 8 sera by absorption chromatography and their agglutinating capacity was studied at pH 7.2 and 5.0. We studied 72 sera from patients with clinical brucellosis taken at the time of hospitalization, 16 from persons in contact with infected animals, and 16 from healthy donors. SAT results at pH 5.0 correlated with those obtained with the Rose Bengal test. Four Rose Bengal-positive sera were found to be SAT-negative at pH 7.2 and SAT-positive at pH 5.0. SAT performed at pH 5.0 with BRUCAPT antigen yielded higher titers than tests performed at pH 7.2 or 5.0 with Ring Test antigen (p IgA fractions were SAT-negative at pH 7.2 and SAT-positive at pH 5.0; the other 5 agglutinated at both pH conditions and were DTT-sensitive. All IgA fractions but one were positive by Rose Bengal. Agglutinating activity of the IgM fraction was not affected by pH. The SAT performed with the buffer and antigen suspension included in the Brucellacapt kit (pH 5.0) is highly useful for detecting agglutinating and non-agglutinating antibodies at pH 7.2.

Application of enzyme linked immunosorbent assays for the diagnosis of bovine brucellosis in Rio Grande do Sul, Brazil

International Nuclear Information System (INIS)

Poester, F.P.; Ramos, E.T.; Thiesen, S.V.

1998-01-01

The results of an indirect ELISA (I-ELISA) and a competitive ELISA (C-ELISA) for detection of antibody to Brucella abortus in cattle were compared with those of conventional serological tests. The sensitivity of I-Elisa using 230 sera from infected animals was 98.6% and the specificity in 720 sera from brucellosis-free animals was 98.2%. The C-ELISA when tested in 94 positive sera and 91 negative sera was 100% sensitive and specific. The relative sensitivity and specificity of I-ELISA compared to C-ELISA in 582 sera from groups of animals vaccinated with S19 by different routes and doses was respectively 64% and 100%. When comparing five serological tests on sera from vaccinated animals, the specificity was 94% for C-ELISA, 93% for 2-mercaptoethanol (2ME), 88% for Rose Bengal Plate test (RBPT), 84% for Slow Tube Agglutination test (SAT) and 34% for I-ELISA. It can be concluded from this study that the RBPT gave less false positive results than the I-ELISA in vaccinated animals and the C-ELISA is a good confirmatory test with the advantage of distinguishing the antibody response due to vaccination from that resulting from infection with Brucella abortus. (author)

Comparison of Rubazyme-M and MACRIA for the detection of rubella-specific IgM

Energy Technology Data Exchange (ETDEWEB)

Best, J.M.; Palmer, S.J.; Morgan-Capner, P.; Hodgson, J. (Saint Thomas' Hospital, London (UK))

1984-02-01

One hundred and eighty-six carefully selected sera were tested for rubella-specific IgM by Rubazyme-M (Abbott Diagnostics) and an M-antibody capture radioimmunoassay (MACRIA). Eleven of these sera were from cases of infectious mononucleosis, six of which gave positive results in MACRIA, while one gave a positive result in Rubazyme-M. Of the remaining 175 sera, 158 gave concordant results whilst 17 sera gave discordant results; these 17 were also tested by serum fractionation. Problems were encountered with all assay systems used. It is therefore recommended that the results of all tests for rubella-specific IgM should be interpreted with caution.

A comparison of Rubazyme-M and MACRIA for the detection of rubella-specific IgM

International Nuclear Information System (INIS)

Best, J.M.; Palmer, S.J.; Morgan-Capner, P.; Hodgson, J.

1984-01-01

One hundred and eighty-six carefully selected sera were tested for rubella-specific IgM by Rubazyme-M (Abbott Diagnostics) and an M-antibody capture radioimmunoassay (MACRIA). Eleven of these sera were from cases of infectious mononucleosis, six of which gave positive results in MACRIA, while one gave a positive result in Rubazyme-M. Of the remaining 175 sera, 158 gave concordant results whilst 17 sera gave discordant results; these 17 were also tested by serum fractionation. Problems were encountered with all assay systems used. It is therefore recommended that the results of all tests for rubella-specific IgM should be interpreted with caution. (Auth.)

An immunochemical method for the quantitation of insulin antibodies

International Nuclear Information System (INIS)

Reeves, W.G.; Kelly, U.

1980-01-01

A 125 I-labelled insulin binding assay is described in which IgG antibody is precipitated by the addition of an optimal concentration of second antibody. Other features include the removal of unlabelled insulin from test sera prior to assay and the use of 22 Na as a volume marker. This approach overcomes problems associated with previous assays for insulin antibodies. Clear differences are seen in the IgG insulin binding capacity (IBC) of sera from patients with insulin resistance and injection site lipo-atrophy when compared with insulin-treated diabetics who lack such complications. The precision and flexibility of this technique make it particularly suitable for studies of the immune response to different species and forms of insulin. (Auth.)

An indirect haemagglutination test for demonstration of gonococcal antibodies using gonococcal pili as antigen. II. Serological investigation of patients attending a dermato-venereological outpatients clinic in Copenhagen

DEFF Research Database (Denmark)

Reimann, K; Lind, I; Andersen, Klaus Ejner

1980-01-01

A total of 1223 serum specimens were obtained from 649 consecutive patients attending a dermatovenereological out-patient clinic in Copenhagen with a request for venereal disease control. The sera were examined for gonococcal antibodies by both a gonococcal complement fixation test (GCF) and an i...

Latex agglutination test (LAT) for the diagnosis of typhoid fever.

Science.gov (United States)

Sahni, Gopal Shankar

2013-06-01

The efficacy of latex agglutination test in the rapid diagnosis of typhoid fever was studied and the result compared with that of blood culture. This study included 80 children suffering from typhoid fever, among which 40 were confirmed by blood culture isolation and 40 had possible typhoid fever based on high Widal's titre (a four-fold rise in the titre of antibody to typhi "O" and "H" antigen was considered as a positive Widal's test result). Eighty children, 40 with febrile illness confirmed to be other than typhoid and 40 normal healthy children were used as negative controls. The various groups were: (i) Study group ie, group I had 40 children confirmed by culture isolation of Salmonella typhi(confirmed typhoid cases). (ii) Control groups ie, (a) group II with 40 febrile controls selected from paediatrics ward where cause other than S typhi has been established, (b) group III with 40 afebrile healthy controls that were siblings of the children admitted in paediatric ward for any reason with no history of fever and TAB vaccination in the last one year, and (c) group IV with 40 children with high Widal's titre in paired sera sample. Widal's test with paired sera with a one week interval between collections were done in all 40 patients. Latex aggtutination test which could detect 900 ng/ml of antigen as observed in checker board titration, was positive in all 40 children from group I who had positive blood culture and in 30 children from group IV who had culture negative and had high Widal's titre positive. Latex agglutination test was positive in 4 children in group II and none in group III. Using blood culture positive cases as true positive and children in groups II and III as true negative, the test had a sensitivity of 100% and specificity of 96%. Latex agglutination test was found to be significantly sensitive (100%) and specific (96%) and could detect 75% more cases in group IV (possible typhoid cases). Thus latex agglutination test can be used for rapid

Humoral leukocyte adherence inhibition (H-LAI) test in screening of high risk group for lung cancer

International Nuclear Information System (INIS)

Kubasova, T.; Koeteles, G.J.; Horvath, M.; Bank, J.

1990-01-01

In screening 150 ore miners a positive humoral immune response to the lung tumor antigen was found in 30 serum samples. Repeated testing of the positive sera (after 1-3 years) was only possible in 15 cases. The reaction of 12 serum samples again was positive, while 2 persons died of lung cancer. The results obtained in these follow-up investigations are discussed. (author). 2 figs., 2 tabs., 11 refs

Una nueva tésera de hospitalidad en territorio cántabro: el oso del castro de Las Rabas (Cervatos, Cantabria

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Fernández Vega, Pedro Ángel

2011-12-01

Full Text Available We present a new tessera hospitalis from the hillfort of Las Rabas (Cervatos, Cantabria. Anepigraphic and with extended bearskin form, its finding reinforces the existence of hospitium among the Cantabrian people and the maintaining of fluid relations with the Plateau and Celtiberia, where we find its closest parallel.Presentamos una nueva tésera de hospitalidad procedente del castro de Las Rabas (Cervatos, Cantabria. Anepígrafa y con forma de piel de oso extendida, su hallazgo refuerza la existencia del hospitium dentro del pueblo cántabro así como el mantenimiento de unas fluidas relaciones con la Meseta y la Celtiberia, en donde encontramos su paralelo más cercano.

Chronic malaria revealed by a new fluorescence pattern on the antinuclear autoantibodies test.

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Benjamin Hommel

Full Text Available BACKGROUND: Several clinical forms of malaria such as chronic carriage, gestational malaria or hyper-reactive malarial splenomegaly may follow a cryptic evolution with afebrile chronic fatigue sometimes accompanied by anemia and/or splenomegaly. Conventional parasitological tests are often negative or not performed, and severe complications may occur. Extensive explorations of these conditions often include the search for antinuclear autoantibodies (ANA. METHODS: We analysed fluorescence patterns in the ANA test in patients with either chronic cryptic or acute symptomatic malaria, then conducted a one-year prospective study at a single hospital on all available sera drawn for ANA detections. We then identified autoantibodies differentially expressed in malaria patients and in controls using human protein microarray. RESULTS: We uncovered and defined a new, malaria-related, nucleo-cytoplasmic ANA pattern displaying the specific association of a nuclear speckled pattern with diffuse cytoplasmic perinuclearly-enhanced fluorescence. In the one-year prospective analysis, 79% of sera displaying this new nucleo-cytoplasmic fluorescence were from patients with malaria. This specific pattern, not seen in other parasitic diseases, allowed a timely reorientation of the diagnosis toward malaria. To assess if the autoantibody immune response was due to autoreactivity or molecular mimicry we isolated 42 autoantigens, targets of malarial autoantibodies. BLAST analysis indicated that 23 of recognized autoantigens were homologous to plasmodial proteins suggesting autoimmune responses directly driven by the plasmodial infection. CONCLUSION: In patients with malaria in whom parasitological tests have not been performed recognition of this new, malaria-related fluorescence pattern on the ANA test is highly suggestive of the diagnosis and triggers immediate, easy confirmation and adapted therapy.

Comparação de dois testes de neutralização, utilizados no diagnóstico sorológico do vesiculovírus Piry Comparison of two neutralization test for the use in the Piry serologic vesiculovirus diagnose

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José Tavares-Neto

1993-10-01

Full Text Available Realizou-se inquérito sorológico para pesquisar anticorpos neutralizantes contra o vesiculovírus Piry, na cidade de Catolândia-Bahia-Brasil. Duas técnicas de vírus-neutralização foram comparadas em cultura das células C6/36, com revelação pelo método imunoenzimático ( TN-C6/36 e em camundongos recém-nascidos (TN-camundongos, que é considerada a prova maior. Em 204 soros, dos 1 .274 colhidos, a concordância das duas técnicas foi de 98,7% (K= 0,9853. Com este resultado do TN-C6/36, que também é mais exeqüível, decidiu-se desenvolver o estudo soro-epidemiológico do vesiculovírus Piry, em Catolândia, baseado nesta nova técnica.A serological survey of Piry was undertaken of the sera of inhabitants of Catolância - Bahia State, Brazil. Serum results obtained by a vesiculovirus neutralization test of C6/36 cells read by ELISA are compared with those obtained by the classic technique, carried out on newbom mice. The agreement between the results was as high as 98.7% of the 204 sera tested and the neutralization test of C6/36 cells was chosen as the most suitable technique for the sero-survey testing.

Detection of IgG against Toxocara in Sera of Employees of Meat Industry

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Alvarado-Esquivel, Cosme; Hernández-Tinoco, Jesús; Sánchez-Anguiano, Luis Francisco; Ramos-Nevárez, Agar; Cerrillo-Soto, Sandra Margarita; Guido-Arreola, Carlos Alberto; Saenz-Soto, Leandro

2015-01-01

Contact with raw meat could represent a risk for Toxocara infection. We assessed the association of Toxocara infection with an occupation of meat worker though a case-control seroprevalence study of 124 meat workers and 248 subjects without this occupation. Sera of participants was analyzed for the presence of anti-Toxocara IgG antibodies. One (0.8%) of the 124 meat workers, and 5 (2.0%) of the 248 controls were positive for anti-Toxocara IgG antibodies (OR=0.39; 95% CI: 0.04-3.41; P=0.66). The seropositive meat worker was a male aged 28 years old, without vision impairment. None of the work characteristics i.e. frequency of contact with raw meat, use of safety practices, history of splashes at face with blood or raw meat, and injuries with sharp material at work was associated with Toxocara exposure. Seroprevalence of Toxocara infection was significantly higher (P=0.04) in meat workers with consumption of boar meat (1/6: 16.7%) than in those without this consumption (0/117: 0%). We conclude that meat workers do not have a higher risk for Toxocara infection than subjects without this occupation do. The 2% seroprevalence of Toxocara infection found in control subjects might suggest a low seroprevalence of this infection among people with other occupations in Durango City. However, additional case-control studies with larger sample sizes to confirm our results are needed. PMID:26508909

A Laboratory-Based Evaluation of Four Rapid Point-of-Care Tests for Syphilis

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Causer, Louise M.; Kaldor, John M.; Fairley, Christopher K.; Donovan, Basil; Karapanagiotidis, Theo; Leslie, David E.; Robertson, Peter W.; McNulty, Anna M.; Anderson, David; Wand, Handan; Conway, Damian P.; Denham, Ian; Ryan, Claire; Guy, Rebecca J.

2014-01-01

Background Syphilis point-of-care tests may reduce morbidity and ongoing transmission by increasing the proportion of people rapidly treated. Syphilis stage and co-infection with HIV may influence test performance. We evaluated four commercially available syphilis point-of-care devices in a head-to-head comparison using sera from laboratories in Australia. Methods Point-of-care tests were evaluated using sera stored at Sydney and Melbourne laboratories. Sensitivity and specificity were calculated by standard methods, comparing point-of-care results to treponemal immunoassay (IA) reference test results. Additional analyses by clinical syphilis stage, HIV status, and non-treponemal antibody titre were performed. Non-overlapping 95% confidence intervals (CI) were considered statistically significant differences in estimates. Results In total 1203 specimens were tested (736 IA-reactive, 467 IA-nonreactive). Point-of-care test sensitivities were: Determine 97.3%(95%CI:95.8–98.3), Onsite 92.5%(90.3–94.3), DPP 89.8%(87.3–91.9) and Bioline 87.8%(85.1–90.0). Specificities were: Determine 96.4%(94.1–97.8), Onsite 92.5%(90.3–94.3), DPP 98.3%(96.5–99.2), and Bioline 98.5%(96.8–99.3). Sensitivity of the Determine test was 100% for primary and 100% for secondary syphilis. The three other tests had reduced sensitivity among primary (80.4–90.2%) compared to secondary syphilis (94.3–98.6%). No significant differences in sensitivity were observed by HIV status. Test sensitivities were significantly higher among high-RPR titre (RPR≥8) (range: 94.6–99.5%) than RPR non-reactive infections (range: 76.3–92.9%). Conclusions The Determine test had the highest sensitivity overall. All tests were most sensitive among high-RPR titre infections. Point-of-care tests have a role in syphilis control programs however in developed countries with established laboratory infrastructures, the lower sensitivities of some tests observed in primary syphilis suggest these would

Low specificity of 2 tetanus rapid tests in Cambodia.

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Schlumberger, M; Yvonnet, B; Lesage, G; Tep, B

2015-01-01

Rapid testing for tetanus on serum or blood allows for an immediate evaluation of individual protection against tetanus in developed countries, using a "single step" immunochromatographic technique using tetanus toxoid. The specificity of these tests, compared to the reference method for tetanus, mouse serum neutralization testing, has however never been assessed in these countries, due to the difficulty to perform serum neutralization titration in mice, because of animal testing bioethical regulations. A collection of sera from adult volunteers in Cambodia, living in rural environment, was tested for tetanus antibodies by ELISA in France, and by mouse serum neutralization in Vietnam. This allowed estimating the sensitivity and specificity of 2 rapid tetanus tests, available on the market: TQS™ and Tetanotop™. The sensitivity of these tests was adequate, compared to mice serum neutralization test, for a test threshold of 0.01 IU/mL, (100% for TQS™, 91% for Tetanotop™), but their specificity was very low (1% for TQS™ and 13% for Tetanotop™). The results prove that these rapid tests for the assessment of individual protection against tetanus should not be used in the adult rural Cambodian population. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

The diagnostic performance evaluation of the SD BIOLINE HIV/syphilis Duo rapid test in southern Ethiopia: a cross-sectional study.

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Shimelis, Techalew; Tadesse, Endale

2015-04-23

To determine the diagnostic performance of the SD BIOLINE HIV/syphilis Duo rapid test. A hospital-based cross-sectional study. This evaluation was conducted at one of the largest hospitals in southern Ethiopia. Serum samples obtained from clients attending the antiretroviral therapy and voluntary counselling and testing centres were used. Sera were originally collected for the purpose of investigating syphilis epidemiology. The performance of the test to detect HIV was evaluated using 400 sera (200 HIV positives and 200 HIV negatives). Also, its performance to detect syphilis was evaluated using 85 syphilis positive and 100 syphilis negative serum samples. Individuals tiebreaker to determine the result. We also used ELISA to resolve discordant HIV results. Syphilis serostatus was determined using the Treponema pallidum haemagglutination assay (TPHA). The respective sensitivity, specificity, positive predictive value and negative predictive value of the SD BIOLINE HIV/syphilis Duo test were 100, 99.5, 99.5 and 100% for HIV and 97.6, 96, 95.4 and 98% for syphilis testing, respectively. In reference to TPHA, the test kit reported 4 false positives and 2 false negative results for syphilis. The κ values were 0.99 for HIV testing and 0.94 for syphilis testing. The excellent performance of the SD BIOLINE HIV/syphilis Duo test to detect HIV as well as syphilis facilitates the integration of syphilis testing and treatment to the already established HIV prevention programme, ultimately contributing to the dual HIV and syphilis elimination goal. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Mast cell activation test in the diagnosis of allergic disease and anaphylaxis.

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Bahri, Rajia; Custovic, Adnan; Korosec, Peter; Tsoumani, Marina; Barron, Martin; Wu, Jiakai; Sayers, Rebekah; Weimann, Alf; Ruiz-Garcia, Monica; Patel, Nandinee; Robb, Abigail; Shamji, Mohamed H; Fontanella, Sara; Silar, Mira; Mills, E N Clare; Simpson, Angela; Turner, Paul J; Bulfone-Paus, Silvia

2018-03-05

Food allergy is an increasing public health issue and the most common cause of life-threatening anaphylactic reactions. Conventional allergy tests assess for the presence of allergen-specific IgE, significantly overestimating the rate of true clinical allergy and resulting in overdiagnosis and adverse effect on health-related quality of life. To undertake initial validation and assessment of a novel diagnostic tool, we used the mast cell activation test (MAT). Primary human blood-derived mast cells (MCs) were generated from peripheral blood precursors, sensitized with patients' sera, and then incubated with allergen. MC degranulation was assessed by means of flow cytometry and mediator release. We compared the diagnostic performance of MATs with that of existing diagnostic tools to assess in a cohort of peanut-sensitized subjects undergoing double-blind, placebo-controlled challenge. Human blood-derived MCs sensitized with sera from patients with peanut, grass pollen, and Hymenoptera (wasp venom) allergy demonstrated allergen-specific and dose-dependent degranulation, as determined based on both expression of surface activation markers (CD63 and CD107a) and functional assays (prostaglandin D 2 and β-hexosaminidase release). In this cohort of peanut-sensitized subjects, the MAT was found to have superior discrimination performance compared with other testing modalities, including component-resolved diagnostics and basophil activation tests. Using functional principle component analysis, we identified 5 clusters or patterns of reactivity in the resulting dose-response curves, which at preliminary analysis corresponded to the reaction phenotypes seen at challenge. The MAT is a robust tool that can confer superior diagnostic performance compared with existing allergy diagnostics and might be useful to explore differences in effector cell function between basophils and MCs during allergic reactions. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights

Development of an Aquaporin-4 Orthogonal Array of Particle-Based ELISA for Neuromyelitis Optica Autoantibodies Detection.

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Francesco Pisani

Full Text Available Serological markers of Nuromyelitis Optica (NMO, an autoimmune disorder of the central nervous system, are autoantibodies targeting the astrocytic water channel aquaporin-4 (AQP4. We have previously demonstrated that the main epitopes for these autoantibodies (AQP4-IgG are generated by the supramolecular arrangement of AQP4 tetramers into an Orthogonal Array of Particles (OAPs. Many tests have been developed to detect AQP4-IgG in patient sera but several procedural issues affect OAP assembly and consequently test sensitivity. To date, the protein based ELISA test shows the lowest sensitivity while representing a valid alternative to the more sensitive cell based assay (CBA, which, however, shows economic, technical and interpretation problems. Here we have developed a high perfomance ELISA in which native OAPs are used as the molecular target. To this aim a native size exclusion chromatography method has been developed to isolate integral, highly pure and AQP4-IgG-recognized OAPs from rat brain. These OAPs were immobilized and oriented on a plastic plate by a sandwich approach and 139 human sera were tested, including 67 sera from NMO patients. The OAP-ELISA showed a 99% specificity and a higher sensitivity (91% compared to the CBA test. A comparative analysis revealed an end-point titer three orders of magnitude higher than the commercial ELISA and six times higher than our in-house CBA test. We show that CNS-extracted OAPs are crucial elements in order to perform an efficient AQP4-IgG test and the OAP-ELISA developed represents a valid alternative to the CBA currently used.

Sphero-echinocytosis of human red blood cells caused by snake, red-back spider, bee and blue-ringed octopus venoms and its inhibition by snake sera.

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Flachsenberger, W; Leigh, C M; Mirtschin, P J

1995-06-01

It was found that bee (Apis mellifera) venom, red-back spider (Latrodectus mactans) venom, blue-ringed octopus (Hapalochlaena maculosa) venom, ten different snake venoms, phospholipase A2 and four snake toxins caused sphero-echinocytosis of human red blood cells at 200 ng/ml. Most venoms and toxins lost the ability to deform human red blood cells when their components of less than mol. wt 10,000 were applied. In a number of cases the sphero-echinocytotic effect was also inhibited by blood sera of Notechis scutatus and Pseudonaja textilis.

Two competitive enzyme immunoassays for the detection of IgG class antibodies to hepatitis a antigen

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Claudia Lamarca Vitral

1991-06-01

Full Text Available Two competitive enzyme immunoassays (EIA techniques were developed: in the first (COMP-1, test sera were added together with HAV antigen on anti-HAV IgG-coated wells followed by an anti-HA VHRP conjugate; in the second (COMP-2, test sera and anti-HA VHRP conjugate competed for HAV epitopes previously adsorbed to anti-HA V IgG-coated wells. Both procedures used tetramethylbenzidine (TMB as a substrate. Both competitive tests were shown to be reproducible and suitable for routine diagnosis and research purposes.Foram desenvolvidos dois ensaios imunoenzimáticos (EIA competitivos: no primeiro (COMP-1 colocou-se numa placa sensibilizada com anti-HAVIgG as amostras teste juntamente como antígeno HA Vea seguir o conjugado anti-HA VHRP; no segundo (COMP-2, as amostras teste e o conjugado anti-HAV HRP competem pelos epitopos do antígeno HAV previamente absorvido na placa sensibilizada do anti-HAV IgG. O substrato utilizado foi tetrametilbenzidina (TMB. Ambas as técnicas mostraram ser produtíveis e aplicáveis para fins de diagnóstico e pesquisa.

Identification and analysis of cytochrome P450IID6 antigenic sites recognized by anti-liver-kidney microsome type-1 antibodies (LKM1).

Science.gov (United States)

Yamamoto, A M; Cresteil, D; Boniface, O; Clerc, F F; Alvarez, F

1993-05-01

Anti-liver-kidney microsome type-1 antibodies (LKM1), present in sera from a group of patients with autoimmune hepatitis, are directed against P450IID6. Previous work, using cDNA constructions spanning most of the P450IID6 protein defined the main immunogenic site between the amino acids (aa), 254-271 and predicted the presence of other putative immunogenic sites in the molecule. Fusion proteins from new cDNA constructions, spanning so-far-untested regions between aa 1-125 and 431-522, were not recognized by LKM1-positive sera. Synthetic peptides, representing sequences from putative immunogenic regions or previously untested regions, allowed a precise definition of four antigenic sites located between peptides 257-269, 321-351, 373-389 and 410-429, which were recognized, respectively, by 14, 8, 1 and 2 out of 15 LKM1-positive sera tested. The minimal sequence of the main antigenic site (peptide 257-269) recognized by the autoantibody was established to be WDPAQPPRD (peptide 262-270). In addition, deletion and replacement experiments showed that aa 263 (Asp) was essential for the binding of the autoantibody to peptide 262-270. Analysis of the second most frequently recognized peptide between aa 321-351, was performed using peptides 321-339 and 340-351 in competitive inhibition studies. Complete elimination of antibody binding to peptide 321-351 obtained by absorption of both shorter peptides indicated that peptide 321-351 is a discontinuous antigenic site. LKM1-positive sera reacting against peptide 321-351 recognized either both the shorter peptides or just one of them preferentially. Results of the present study suggest that the production of LKM1 antibodies is an antigen-driven, poly- or oligoclonal B cell response. The identification of antigenic sites will allow: (i) the development of specific diagnostic tests and (ii) further studies on the pathogenic value of LKM1 antibodies in autoimmune hepatitis.

Common antigenic determinants of haemoglobin as basis of immunological cross-reactivity between chironomid species (Diptera, Chironomidae): studies with human and animal sera.

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Baur, X; Dewair, M; Haegele, K; Prelicz, H; Scholl, A; Tichy, H

1983-01-01

Chironomids, of which approximately 10,000 species exist, are reported to cause severe immediate type allergic diseases in man. In the present study, immunological cross-reactivity between 14 chironomid species from different continents was proven by RAST inhibition, double immunodiffusion and a new allergoprint technique, based upon PAGE separation of insect crude extracts. Using isolated chironomid haemoglobins and sera of sensitized persons, as well as rabbit antibodies against larval crude extract or against the haemoglobin fraction of Chironomus thummi, it could be proven that cross-reactivity derives at least predominantly from haemoglobin components with common antigenic determinants in the different species. Images Fig. 2 Fig. 7 Fig. 8 Fig. 9 PMID:6197219

Evaluation of an enzyme linked immunosorbent assay kit for the detection of Babesia bovis antibodies in cattle in Argentina

Energy Technology Data Exchange (ETDEWEB)

Echaide, S; Echaide, I E; Mangold, A J; Lugaresi, C I; Guglielmone, A A [Estacion Experimental Agropecuaria, Instituto Nacional de Tecnologia Agropecuaria, Rafaela, Santa Fe (Argentina); Gaido, A B [Estacion Experimental Agropecuaria Salta, Salta (Argentina)

1998-11-01

An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to Babesia bovis was evaluated by using sera of 874 cattle carrying B. bovis antibodies, 700 sera of uninfected cattle, and 357 sera from calves from 16 herds subjected to different B. bovis inoculation rates. The seropositive/ seronegative cut-off point set as double the mean percent positivity of negative cattle sera (= 16%). The sensitivity of the ELISA (four trials) ranged from 97.1% to 100% and the specificity (three trials) varied from 92.0% to 97.0%. The agreement between ELISA and immunofluorescent antibody test was {>=} 90.0% in 18 of 23 evaluations and it ranged from 86.0% to 88.0% in the remainder. The correlation coefficient between percentage of sera positive to ELISA and IFA test in 16 herds was 0.9958 (P<0.001). The ELISA has the advantages of a high sensitivity, objectivity and capacity to test large number of samples in short period of time and could replace the IFA test specially for epidemiological studies. (author) 11 refs, 1 fig., 2 tabs

Evaluation of an enzyme linked immunosorbent assay kit for the detection of Babesia bovis antibodies in cattle in Argentina

International Nuclear Information System (INIS)

Echaide, S.; Echaide, I.E.; Mangold, A.J.; Lugaresi, C.I.; Guglielmone, A.A.; Gaido, A.B.

1998-01-01

An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to Babesia bovis was evaluated by using sera of 874 cattle carrying B. bovis antibodies, 700 sera of uninfected cattle, and 357 sera from calves from 16 herds subjected to different B. bovis inoculation rates. The seropositive/ seronegative cut-off point set as double the mean percent positivity of negative cattle sera (= 16%). The sensitivity of the ELISA (four trials) ranged from 97.1% to 100% and the specificity (three trials) varied from 92.0% to 97.0%. The agreement between ELISA and immunofluorescent antibody test was ≥ 90.0% in 18 of 23 evaluations and it ranged from 86.0% to 88.0% in the remainder. The correlation coefficient between percentage of sera positive to ELISA and IFA test in 16 herds was 0.9958 (P<0.001). The ELISA has the advantages of a high sensitivity, objectivity and capacity to test large number of samples in short period of time and could replace the IFA test specially for epidemiological studies. (author)

Inhibtion of the mixed leukocyte reaction by alloantisera in man

International Nuclear Information System (INIS)

Jonker, M.; Rood, J.J. van

1977-01-01

Some technical aspects of MLC (mixed leukocyte culture) inhibition by sera obtained from multiparous women were studied. The variability of the MLC response was very high. The serum source in the cultures was probably to a large extent responsible for this variability. A specific inhibition, which was observed with some test sera, could be removed by dialysis against PBS (phosphate buffered saline). To make the evaluation of inhibition by immune sera objective, a scoring system was introduced for the degree of inhibition. Test sera were usually added to the cultures. Alternatively, stimulator and responder cells were preincubated with the test serum. Preincubation of the stimulator cells did not show a difference in inhibition pattern when this was compared with serum addition. Preincubation of the responder cells showed a completely different inhibition pattern. The MLC inhibition test compared very well with an indirect immunofluorescence test and a cytotoxicity test using B-cell enriched cell suspensions. (author)

Indirect immunofluorescence assay for the simultaneous detection of antibodies against clinically important old and new world hantaviruses.

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Sabine Lederer

Full Text Available In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV, Puumala (PUUV, Seoul (SEOV, Saaremaa (SAAV, Dobrava (DOBV, Sin Nombre (SNV or Andes (ANDV. For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n=97; SEOV (China, n=5; DOBV (Romania, n=7; SNV (Canada, n=23; ANDV (Argentina and Chile, n=52. The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n=177; 96% and/or IgM (n=131; 72%, while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99% of the patient sera were IgG and 131 (71% IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%. Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%. Of the 89 control sera, 2 (2% showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV.

A large-scale radiometric micro-quantitative complement fixation test for serum antibody titration

International Nuclear Information System (INIS)

Bengali, Z.H.; Levine, P.H.; Das, S.R.

1980-01-01

A micro-quantitative complement fixation (CF) procedure based on 51 Cr release is described. The method employs 50% hemolysis as end point and the alternation equation to calculate the amount of complement involved in the hemolytic reaction. Compared to the conventional CF tests, the radiometric procedure described here is very precise and consistently reproducible. Also, since only 3 4-fold dilutions of sera are used for the titration of antibodies over a wide range of concentrations, the test is very concise and is economical to perform. Its format is amenable to automation and computerization. This radioimetric CF procedure is thus most useful for large-scale immunological research and epidemiological surveilance studies. (Auth.)

Comparative evaluation of antibody positive titer by ELISA and IFA in Theileria annulata vaccinated cattle in Iran

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Hashemi-Fesharki R.

2006-03-01

Full Text Available An enzyme linked immunosorbent assay (ELISA was used to evaluate antibody positive titer in vaccinated and non-vaccinated cattle using schizont infected myeloid cells as an antigen. The result was compared with indirect fluorescent antibody level in the same animals. For this study 116 milking cows, 95 vaccinated and 21 non-vaccinated, were bleeded in order to prepare sera. They were tested with both ELISA and IFA tests. 94 sera had positive antibody titer and 22 sera were negative through ELISA test but, with IFA test, only 89 sera showed positive antibody titer and 27 were negative. Thereby, it was concluded that the sensitivity and specificity of ELISA test in comparison with IFA test was 95.5 % and 66.6 % respectively. This study generally indicated that ELISA could be an effective test for seroepidemiological investigations of bovine tropical theileriosis, and it is considered to be valid as an additional test to distinguish the vaccinated from the non vaccinated cattle in order to schedule vaccination programs.

Predictive factors for the development of diabetes in women with previous gestational diabetes mellitus

DEFF Research Database (Denmark)

Damm, P.; Kühl, C.; Bertelsen, Aksel

1992-01-01

OBJECTIVES: The purpose of this study was to determine the incidence of diabetes in women with previous dietary-treated gestational diabetes mellitus and to identify predictive factors for development of diabetes. STUDY DESIGN: Two to 11 years post partum, glucose tolerance was investigated in 241...... women with previous dietary-treated gestational diabetes mellitus and 57 women without previous gestational diabetes mellitus (control group). RESULTS: Diabetes developed in 42 (17.4%) women with previous gestational diabetes mellitus (3.7% insulin-dependent diabetes mellitus and 13.7% non...... of previous patients with gestational diabetes mellitus in whom plasma insulin was measured during an oral glucose tolerance test in late pregnancy a low insulin response at diagnosis was found to be an independent predictive factor for diabetes development. CONCLUSIONS: Women with previous dietary...

HIV risk perception and testing behaviours among men having sex with men (MSM) reporting potential transmission risks in the previous 12 months from a large online sample of MSM living in Germany.

Science.gov (United States)

Marcus, Ulrich; Gassowski, Martyna; Drewes, Jochen

2016-10-22

HIV testing and serostatus awareness are essential to implement biomedical strategies (treatment as prevention; oral chemoprophylaxis), and for effective serostatus-based behaviours (HIV serosorting; strategic positioning). The analysis focuses on the associations between reported sexual risks, the perceived risk for HIV infection, and HIV testing behaviour in order to identify the most relevant barriers for HIV test uptake among MSM living in Germany. MSM were recruited to a nationwide anonymous online-survey in 2013 on MSM social networking/dating sites. Questions covered testing behaviours, reasons for testing decisions, and HIV risk perception (5-point scale). Additional questions addressed arguments in favour of home/ home collection testing (HT). Using descriptive statistics and logistic regression we compared men reporting recent HIV testing (RT; previous 12 month) with men never tested (NT) in a subsample not previously diagnosed with HIV and reporting ≥2 episodes of condomless anal intercourse (CLAI) with a non-steady partner of unknown HIV serostatus in the previous 12 months. The subsample consisted of 775 RT (13 % of RT) and 396 NT (7 % of NT). The number of CLAI episodes in the last 12 months with non-steady partners of unknown HIV status did not differ significantly between the groups, but RT reported significantly higher numbers of partners (>5 AI partners: 65 vs. 44 %). While perceived risks regarding last AI were comparable between the groups, 49vs. 30 % NT were risks (67 %) and routine testing (49 %) were the most common testing reasons for RT, while the strong belief not to be infected (59 %) and various worries (41 %) and fears of testing positive (35 %) were predominant reasons of NT. Greater anonymity (aOR 3.2; 2.4-4.4), less embarrassment, (aOR 2.8; 1.9-4.1), and avoiding discussions on sexual behaviour (aOR 1.6; 1.1-2.2) were emphasized in favour of HT by NT. Perceived partner knowledge and reasons reflecting perceived gay- and

Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey.

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Tel, O Y; Arserim, N B; Keskin, O

2011-12-01

In order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701) as antigen. Eighty (11.7%) sera from horses and 9 (11.5%) sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9%) horses in Sanliurfa Province and 24 (8.7%) horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5% of female (n=343) and 10.1% of male (n = 59) horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4%, n=114) were seropositive than females (6.7%, n=163). Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5%) were positive by ELISA. Among the 9 positive sera, 6 (12.8%) were from donkeys 1-5 years old and 3 (13.6%) were from donkeys >5 years of age. No positive sera were found in donkeys less than 1 year old. Five (12.5%) sera of females and 4 (10.5%) sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey

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O. Y. Tel

2011-05-01

Full Text Available n order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701 as antigen. Eighty (11.7 % sera from horses and 9 (11.5 % sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9 % horses in Sanliurfa Province and 24 (8.7 % horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5 % of female (n = 343 and 10.1 % of male (n = 59 horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4 %, n = 114 were seropositive than females (6.7 %, n = 163. Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5 % were positive by ELISA. Among the 9 positive sera, 6 (12.8 % were from donkeys 1-5 years old and 3 (13.6 % were from donkeys >5years of age. Nopositive sera were found in donkeys less than 1 year old. Five(12.5 % sera of females and 4(10.5 % sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

The tryptic cleavage product of the mature form of the bovine desmoglein 1 ectodomain is one of the antigen moieties immunoprecipitated by all sera from symptomatic patients affected by a new variant of endemic pemphigus.

Science.gov (United States)

Abréu-Vélez, Ana María; Javier Patiño, Pablo; Montoya, Fernando; Bollag, Wendy B

2003-01-01

Multiple antigens are recognized by sera from patients with pemphigus foliaceus (PF). Several have been identified including keratin 59, desmocollins, envoplakin, periplakin, and desmogleins 1 and 3 (Dsg1 and Dsg3). In addition, an 80 kDa antigen was identified as the N-terminal fragment of Dsg1 using as antigen source an insoluble epidermal cell envelope preparation. However, still unsolved was the identity of the most important antigenic moiety, a 45 kDa tryptic fragment which is recognized by all sera from patients with fogo selvagem, pemphigus foliaceus, by half of pemphigus vulgaris sera and by a new variant of endemic pemphigus in E1 Bagre, Colombia that resembles Senear-Usher syndrome. Here, we report the identification of the 45 kDa conformational epitope of a soluble tryptic cleavage product from viable bovine epidermis. To elucidate the nature of this peptide, viable bovine epidermis was trypsin-digested, and glycosylated peptides were partially purified on a concanavalin A (Con-A) affinity column. This column fraction was then used as an antigen source for further immunoaffinity purification. A PF patient's serum covalently coupled to a Staphylococcus aureus protein A column was incubated with the Con-A eluted products and the immuno-isolated antigen was separated by SDS-PAGE, transferred to a membrane, and visualized with Coomassie blue, silver and amido black stains. The 45 kD band was subjected to amino acid sequence analysis revealing the sequence, EXIKFAAAXREGED, which matched the mature form of the extracellular domain of bovine Dsg1. This study confirms the biological importance of the ectodomain of Dsg1 as well as the relevance of conformational epitopes in various types of pemphigus.

Investigation of a panel of monoclonal antibodies and polyclonal sera against anthrax toxins resulted in identification of an anti-lethal factor antibody with disease-enhancing characteristics.

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Kulshreshtha, Parul; Tiwari, Ashutosh; Priyanka; Joon, Shikha; Sinha, Subrata; Bhatnagar, Rakesh

2015-12-01

Hybridomas were created using spleen of mice that were actively immunized with rLFn (recombinant N-terminal domain of lethal factor). Later on, separate group of mice were immunized with rLFn to obtain a polyclonal control for passive immunization studies of monoclonal antibodies. This led to the identification of one cohort of rLFn-immnized mice that harboured disease-enhancing polyclonal antibodies. At the same time, the monoclonal antibodies secreted by all the hybridomas were being tested. Two hybridomas secreted monoclonal antibodies (H10 and H8) that were cross-reactive with EF (edema factor) and LF (lethal factor), while the other two hybridomas secreted LF-specific antibodies (H7 and H11). Single chain variable fragment (LETscFv) was derived from H10 hybridoma. H11 was found to have disease-enhancing property. Combination of H11 with protective monoclonal antibodies (H8 and H10) reduced its disease enhancing nature. This in vitro abrogation of disease-enhancement provides the proof of concept that in polyclonal sera the disease enhancing character of a fraction of antibodies is overshadowed by the protective nature of the rest of the antibodies generated on active immunization. Copyright © 2015. Published by Elsevier Ltd.

Standardization of micro-enzyme-linked immunosorbent assay (ELISA) and Western blot for detection of Trypanosoma cruzi antibodies using extracts from Mexican strains as antigens.

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Sánchez, B; Monteón, V; Reyes, P A; Espinoza, B

2001-01-01

This report describes two assays for the detection of anti-Trypanosoma cruzi antibodies using Mexican strains of the parasite and the concordance with two assays previously evaluated at the Instituto Nacional de Cardiología Ignacio Chávez in Mexico City. Micro-enzyme-linked immunosorbent assay (ELISA) and Western blot were used for the detection of T. cruzi antibodies with a total extract of epimastigote from Ninoa and Queretaro, which are Mexican strains of T. cruzi. To standardize these methods, a total of 246 serum samples was used. In addition, sera from six confirmed Mexican chronic individuals in the asymptomatic phase were also used for comparison with the Argentinean antigen. ELISA was 100% specific in that no false positive results were found with sera of both healthy individuals and non-Chagasic cardiopaths. Sera from individuals infected with Leishmania sp. showed approximately 16% of cross-reaction with ELISA. The test showed a positive predictive value of 90% and a negative predictive value of 100%. Western blot was also a highly sensitive test for detecting chronic Chagasic symptomatic patients from Mexico because no false negative results were obtained. Furthermore, it was possible to use Western blot to detect seven immunodominant antigens of approximately 30, 32, 40, 42, 65, 70, and 83 kDa. Concordance with two previous standardized tests at the Instituto Nacional de Cardiología showed a Kappa index of 0.96, indicating high concordance between the results obtained at these two laboratories. Finally, ELISA using Ninoa antigen extract was more sensitive than ELISA with an Argentinean extract, which failed to detect individuals in the chronic asymptomatic phase (undetermined phase) of infection. This study indicates that ELISA and Western blot using Ninoa and/or Queretaro extracts of T. cruzi as antigens are useful tools in the detection of individuals who have been exposed to T. cruzi both in the undetermined/asymptomatic and symptomatic phases

Serological Evidence of Contrasted Exposure to Arboviral Infections between Islands of the Union of Comoros (Indian Ocean.

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Koussay Dellagi

2016-12-01

Full Text Available A cross sectional serological survey of arboviral infections in humans was conducted on the three islands of the Union of Comoros, Indian Ocean, in order to test a previously suggested contrasted exposure of the three neighboring islands to arthropod-borne epidemics. Four hundred human sera were collected on Ngazidja (Grande Comore, Mwali (Mohéli and Ndzouani (Anjouan, and were tested by ELISA for IgM and/or IgG antibodies to Dengue (DENV, Chikungunya (CHIKV, Rift Valley fever (RVFV, West Nile (WNV, Tick borne encephalitis (TBEV and Yellow fever (YFV viruses and for neutralizing antibodies to DENV serotypes 1-4. Very few sera were positive for IgM antibodies to the tested viruses indicating that the sero-survey was performed during an inter epidemic phase for the investigated arbovirus infections, except for RVF which showed evidence of recent infections on all three islands. IgG reactivity with at least one arbovirus was observed in almost 85% of tested sera, with seropositivity rates increasing with age, indicative of an intense and long lasting exposure of the Comorian population to arboviral risk. Interestingly, the positivity rates for IgG antibodies to DENV and CHIKV were significantly higher on Ngazidja, confirming the previously suggested prominent exposure of this island to these arboviruses, while serological traces of WNV infection were detected most frequently on Mwali suggesting some transmission specificities associated with this island only. The study provides the first evidence for circulation of RVFV in human populations from the Union of Comoros and further suggests that the virus is currently circulating on the three islands in an inconspicuous manner. This study supports contrasted exposure of the islands of the Comoros archipelago to arboviral infections. The observation is discussed in terms of ecological factors that may affect the abundance and distribution of vector populations on the three islands as well as concurring

Comparison between an immunochromatographic test with an amplified ELISA for detecting e antigen and anti-e antigen antibodies in chronic Hepatitis B

International Nuclear Information System (INIS)

Mainet Gonzalez, Damian; Palenzuela Gardon, Daniel O; Aguilar Rubido; Julio C

2009-01-01

The disappearance of the e antigen and the appearance of anti-e antigen antibodies are two biomarkers that indicate favorable prognosis in Hepatitis B. In this study the Advanced QualityTM immunochromatographic test for detecting those biomarkers was compared to the Vidas semi-quantitative ELISA test. Our hypothesis was that it is possible to use these biomarkers measured in a rapid and simple Advanced QualityTM immunochromatographic test for evaluating the therapeutic response in clinical trials with chronic hepatitis B patients. The two methods were done following the manufacturer's instructions. The sera were taken from 69 patients with chronic hepatitis B of the clinical trial of the CIGB 440 therapeutic candidate. The immunochromatographic test and ELISA for detecting e antigen and anti-e antigen antibodies presented from substantial to almost perfect agreement in the evaluation of the sera of chronic Hepatitis B patients in a clinical trial. The immunochromatographic test for detecting e antigen had a low positive average agreement and a high negative average agreement compared to the ELISA. Nevertheless, the immunochromatographic test for detecting anti-e antigen antibodies had a high negative and positive average agreement in comparison to the ELISA. The immunochromagraphic test for the e antigen had a lower positive average agreement compared to the ELISA and some patients infected with Hepatitis B virus could not be detected by the former assay. The immunochromatographic test for anti-e antigen antibodies showed a similar performance to that of ELISA and could therefore be used in clinical trials for chronic Hepatitis B in health institutions without the need of a highly qualified lab technician. (author)

Development and validation of a quantitative competitive ELISA for potency testing of equine anti rabies sera with other potential use.

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Korimbocus, Jehanara; Dehay, Nicolas; Tordo, Noël; Cano, François; Morgeaux, Sylvie

2016-06-14

In case of a bite by a rabies infected animal, the World Health Organisation recommends a prophylactic treatment including the administration of Human Rabies Immunoglobulins (HRIGs) or highly purified F(ab')2 fragments produced from Equine Rabies Immunoglobulin (F(ab')2 - ERIGs). According to international regulation, quality control of F(ab')2 - ERIGs lots requires potency testing by the in vivo Mouse Neutralisation Test (MNT) prior marketing. However, the strategy of the 3Rs (Reduce, Refine, Replace) for animal testing required by the European Directive encourages the replacement of the in vivo potency test by an in vitro assay. In this context, a competitive ELISA method (c-ELISA) has been developed by the Agence Nationale de Sécurité du Médicament et des Produits de Santé where F(ab')2 - ERIGs are in competition with a monoclonal antibody recognizing the trimeric native form of the rabies glycoprotein. After a full validation study, the c-ELISA has been applied to commercial batches of F(ab')2 - ERIGs. A correlation study with the MNT demonstrated a similarity between the two methods (r=0.751). Moreover, the c-ELISA method which does not need any species specific reagent has been applied to HRIGs potency testing as an alternative method to Rapid Fluorescent Focus Inhibition Test (RFFIT), thus avoiding the handling of live rabies virus in BSL3 containment. In conclusion, the c-ELISA has shown its potential to replace MNT and possibly RFFIT for the quantification of rabies immunoglobulin. After optimisation it may be used for the quantification of rabies immunoglobulin in any animal species, notably for rabies immunogenicity assay in mice. Copyright © 2016 Elsevier Ltd. All rights reserved.

Evaluation of an antibody-detection ELISA using pre-coated plates in the diagnosis of Trypanosoma congolense and T. vivax infections in cattle in Mali

International Nuclear Information System (INIS)

Diall, O.; Bocoum, Z.; Sanogo, Y.

2000-01-01

Under the Coordinated Research Programme ''Use of immunoassay for improved diagnosis of trypanosomosis and monitoring trypanosomosis control programmes'', a new ELISA test has been validated. The test was designed for trypanosome specific antibody detection. The purpose of the work was to study the main parameters of the test (sensitivity and specificity) using reference sera provided by IAEA and field sera collected in Mali. The field sera were collected in different ecological zones and included sera from trypanosomosis endemic areas as well as sera from tsetse free areas. The test was performed according to the protocol proposed by IAEA. The results have shown a poor stability of the reference sera for both T. congolense and T. vivax. By using de-ionized water, we got better OD's for T. vivax C++ reference serum, while the T. congolense C++ had to be replaced by a locally produced reference serum. Under the above conditions, the T. congolense system gave quite encouraging results (sensitivity: 100%; specificity: 87.5-95%). This system was able to differentiate quite well animals from tsetse infected areas from those originating from tsetse free areas and could therefore be used in epidemiological surveys. The T. vivax system showed a good sensitivity (100%), but a poor specificity (37.5-57.5%). The T. vivax system could be improved by establishing a cut-off point based on local negative populations. (author)

Fundamental study of Triosorb M-125

International Nuclear Information System (INIS)

Kasama, Jun; Shindo, Toshihiko; Ohishi, Masao; Kusumoto, Toshihiro; Yoshimura, Masami

1975-01-01

The usefullness of the Triosorb M test for measuring thyroid function indirectly was evaluated in 82 pregnant women and subjects with or without thyroid abnormalities. Satisfactory reproducibility was obtained in intraassays and interassays; coefficient variation were +-1.71% and +-2.44% respectively. Triosorb M values were affected by changes of incubation time and temperature, especially by the latter. Triosorb M values dropped significantly in hyperthyroid sera as the incubation temperature rose, while values rose slightly in hypothyroid sera. Incubation time had almost the same effect on both hyper- and hypothyroid sera values. A good correlation was observed between Triosorb M values and Triosorb values, when incubated at 20 0 C or 25 0 C for 20 min. Incubated at 25 0 C, Triosorb M values were lower than those of Triosorb tests on the same hyperthyroid sera. Incubated at 20 0 C, both tests showed very similar values in hyperthyroid sera as well as in hypothyroid and normal sera. In normal subjects, Triosorb M values were 27.9+-2.3% with a rare overlap with those of hyperthyroid, and a moderate overlap with those of hypothyroid. The Triosorb M test requires only 0.5 ml of serum and can be carried on in a short time with strict controlling of the incubation temperature and time, and might be one of the useful examinations of thyroid function. (auth.)

Antigenisitas, Sensitivitas, dan Spesifisitas Protein Toxocara canis pada Pemeriksaan Antibodi Serum Mencit dengan Indirect-ELISA

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Sri Subekti Bendryman

2015-05-01

Full Text Available The aim of this research were to determine antigenicity, sensitivity, and specificity of Toxocara canisprotein used as antigen in indirect-ELISA for the detection antibody against the worm in the infected hostin order to proper diagnose kit. The design used was true experimental, with Post-test Only ControlGroups Design. Mouse was immunized with various worm homogenates used to antigenicity, sensitivityand specificity tests of T. canis protein with indirect-ELISA technique. The independence variable werevarious immunogens (homogenates; the dependence variables were antigenicity, sensitivity and specificityvalues interpreted by optical density (OD value of mouse sera; and controlled variable were mouse strain,feed and retrieval time of sera. The result showed that OD values of mouse sera immunized with T. canisand T.cati homogenate were signicantly difference (p<0.01 as compared to those immunized withAncylostoma spp., Dipylidium caninum and control sera. Using the diagnosis based on the finding ofToxocara, the sensitivity of OD value by ELISA result from mouse sera immunized with Toxocara spp.homogenate were 100%. Using negative OD value by ELISA from mouse sera immunized with Ancylostomaspp. and D. caninum homogenate, the specificity of the test was 87.5%. In conclusion, protein of T.canishas the same antigenicity against anti-T. canis and anti-T. cati sera, but they had the lower antigenicityagainst anti-Ancylostoma spp. and anti-D.caninum sera. As the sensitivity value of 100% and specificityvalue of 87.5%, in detecting antibody against toxocariasis, the possibility of obtaining false positive was12.5%.

Combined rapid (TUBEX test for typhoid-paratyphoid A fever based on strong anti-O12 response: design and critical assessment of sensitivity.

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Meiying Yan

Full Text Available Rapid diagnostics can be accurate but, often, those based on antibody detection for infectious diseases are unwittingly underrated for various reasons. Herein, we described the development of a combined rapid test for two clinically-indistinguishable bacterial diseases, typhoid and paratyphoid A fever, the latter fast emerging as a global threat. By using monoclonal antibodies (mAbs to bacterial antigens of known chemical structures as probes, we were able to dissect the antibody response in patients at the level of monosaccharides. Thus, a mAb specific for a common lipopolysaccharide antigen (O12 found in both the causative organisms was employed to semi-quantify the amounts of anti-O12 antibodies present in both types of patients in an epitope-inhibition particle-based (TUBEX immunoassay. This colorimetric assay detected not only anti-O12 antibodies that were abundantly produced, but also, by steric hindrance, antibodies to an adjoining epitope (O9 or O2 in the typhoid or paratyphoid bacillus, respectively. Sensitivity and, particularly, reaction intensities, were significantly better than those obtained using an anti-O9 or anti-O2 mAb-probe in the examination of paired sera from 22 culture-confirmed typhoid patients (sensitivity, 81.8% vs 75.0% or single sera from 36 culture-confirmed paratyphoid patients (52.8% vs 28.6, respectively. Importantly, sensitivity was better (97.1% for typhoid, 75.0% for paratyphoid if allowance was made for the absence of relevant antibodies in certain specimens as determined by an independent, objective assay (ELISA--such specimens might have been storage-denatured (especially the older paratyphoid samples or procured from non-responders. Benchmarking against ELISA, which revealed high concordance between the two tests, was useful and more appropriate than comparing with culture methods as traditionally done, since antibody tests and culture target slightly different stages of these diseases. Paired sera

Cytokine profile and nitric oxide levels in sera from patients with brucellosis

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Refik M.

2004-01-01

Full Text Available The aims of this study were to investigate the serum levels of some cytokines [tumor necrosis factor-alpha (TNF-alpha, interleukin 1ß (IL-1ß, IL-2R, IL-6, and IL-8] and nitric oxide (NO levels in patients with untreated brucellosis and to test the correlation of these parameters with each other. The study was conducted on 67 subjects, 37 patients with brucellosis and 30 healthy individuals with no history of Brucella infection. Brucellosis was identified by a positive blood culture and/or increased Brucella antibodies in serological tests in addition to compatible clinical symptoms. Cytokine profile analysis was performed by the immulite chemiluminescent enzyme immunometric assay whose inter- and intra-assay coefficients of variance were 2.6-3.6 and 4.4-8.5%, respectively. The levels of nitrites/nitrates, which are representative of NO levels, were measured by the Griess method. Patients with brucellosis had significantly elevated serum levels of nitrites/nitrates, IL-2R, IL-6 and IL-8 (mean ± SD, 102.8 ± 23.8 µmol/l, 806.1 ± 58.5 U/ml, 21.1 ± 2.3 pg/ml, and 8.8 ± 1.6 pg/ml, respectively compared to healthy controls, whereas TNF-alpha and IL-1ß levels were unchanged. No statistically significant correlation was detected between any of the studied cytokine levels and nitrate/nitrite concentrations according to Pearson's linear correlation test. We conclude that only IL-6, IL-8 and IL-2R are elevated in brucellosis and the extent of elevation depends on the severity and clinical pattern of the disease. Moderate elevation in serum NO was comparable to that observed in previous studies. This explains the absence or very rare occurrence of septic shock in brucellosis.

Human antibody fragments specific for Bothrops jararacussu venom reduce the toxicity of other Bothrops sp. venoms.

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Roncolato, Eduardo Crosara; Pucca, Manuela Berto; Funayama, Jaqueline Carlos; Bertolini, Thaís Barboza; Campos, Lucas Benício; Barbosa, José Elpidio

2013-01-01

Approximately 20,000 snakebites are registered each year in Brazil. The classical treatment for venomous snakebite involves the administration of sera obtained from immunized horses. Moreover, the production and care of horses is costly, and the use of heterologous sera can cause hypersensitivity reactions. The production of human antibody fragments by phage display technology is seen as a means of overcoming some of these disadvantages. The studies here attempted to test human monoclonal antibodies specific to Bothrops jararacussu against other Bothrops sp. venoms, using the Griffin.1 library of human single-chain fragment-variable (scFv) phage antibodies. Using the Griffin.1 phage antibody library, this laboratory previously produced scFvs capable of inhibiting the phospholipase and myotoxic activities of Bothrops jararacussu venom. The structural and functional similarities of the various forms of phospholipase A2 (PLA₂) in Bothrops venom served as the basis for the present study wherein the effectiveness of those same scFvs were evaluated against B. jararaca, B. neuwiedi, and B. moojeni venoms. Each clone was found to recognize all three Bothrops venoms, and purified scFvs partially inhibited their in vitro phospholipase activity. In vivo assays demonstrated that the scFv clone P2B7 reduced myotoxicity and increased the survival of animals that received the test venoms. The results here indicate that the scFv P2B7 is a candidate for inclusion in a mixture of specific antibodies to produce a human anti-bothropic sera. This data demonstrates that the human scFv P2B7 represents an alternative therapeutic approach to heterologous anti-bothropic sera available today.

Allergy to fish collagen: Thermostability of collagen and IgE reactivity of patients' sera with extracts of 11 species of bony and cartilaginous fish.

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Kobayashi, Yukihiro; Kuriyama, Takuma; Nakagawara, Ryoko; Aihara, Michiko; Hamada-Sato, Naoko

2016-10-01

Parvalbumin was identified as a major fish allergen, and has been well investigated. Collagen was identified as a second allergen; however, its allergenic properties remain uncharacterized. Although fish is an important staple in coastal countries, its thermostability is unknown. Therefore, we aimed to determine the thermostability of fish collagen as an allergen. Meat of seven bony and four cartilaginous fishes was heated at various temperatures and times, and extracts were analyzed using SDS-PAGE, IgE-ELISA, and SPTs. Collagen was dissolved from heated meat of Pacific mackerel into a crude extract. Collagen in the extracts was degraded at a high heating load-140 °C (10 min) or 100 °C (320 min). However, ELISA revealed the IgE reactivities of patients' sera with the extracts were unchanged even after heating the samples. Patients strongly reacted to extract proteins of other bony fish, which were detected by patients' IgE even after heating at 100 °C (320 min). In contrast, reactivities of the extracts of cartilaginous fish were lower than those of bony fish. SPTs in one patient revealed that all bony and cartilaginous fish extracts prepared from heated meat elicited allergic reactions. The IgE reactivity of patients' sera to fish collagen in extracts was retained even when fish meat was treated by a high heating load. As for the fish collagen, the IgE reactivities to cartilaginous fish were lower than that to bony fish. Reducing IgE reactivity to fish meat using heat is difficult, and other modalities will be required to produce hypoallergenic fish meat. Copyright © 2016 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.

Female Sexual Dysfunction in the Late Postpartum Period Among Women with Previous Gestational Diabetes Mellitus.

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Sargin, Mehmet Akif; Yassa, Murat; Taymur, Bilge Dogan; Taymur, Bulent; Akca, Gizem; Tug, Niyazi

2017-04-01

To compare the status of female sexual dysfunction (FSD) between women with a history of previous gestational diabetes mellitus (GDM) and those with follow-up of a healthy pregnancy, using the female sexual function index (FSFI) questionnaire. Cross-sectional study. Department of Obstetrics and Gynecology, Fatih Sultan Mehmet Training and Research Hospital, Istanbul, Turkey, from September to December 2015. Healthy sexually active adult parous females were included. Participants were asked to complete the validated Turkish versions of the FSFI and Hospital Anxiety and Depression Scale (HADS) questionnaires. Student's t-test was used for two-group comparisons of normally distributed variables and quantitative data. Mann-Whitney U-test was used for two-group comparisons of non-normally distributed variables. Pearson's chi-squared test, the Fisher-FreemanHalton test, Fisher's exact test, and Yates' continuity correction test were used for comparison of qualitative data. The mean FSFI scores of the 179 participants was 23.50 ±3.94. FSFI scores and scores of desire, arousal, lubrication, orgasm, satisfaction, and pain were not statistically significantly different (p>0.05), according to a history of GDM and types of FSD (none, mild, severe). HADS scores and anxiety and depression types did not statistically significantly differ according to the history of GDM (p>0.05). An association could not be found in FSFI scores between participants with both the history of previous GDM and with healthy pregnancy; subclinical sexual dysfunction may be observed in the late postpartum period among women with a history of previous GDM. This may adversely affect their sexual health.

Screening for transfusion transmissible infections using rapid diagnostic tests in Africa: a potential hazard to blood safety?

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Prugger, C.; Laperche, S.; Murphy, E. L.; Bloch, E. M.; Kaidarova, Z.; Tafflet, M.; Lefrère, J.-J.; Jouven, X.

2016-01-01

Rapid diagnostic tests (RDTs) are routinely used in African blood centres. We analysed data from two cross-sectional studies representing 95 blood centres in 29 African countries. Standardized panels of sera containing varying concentrations of anti-human immunodeficiency virus (HIV) antibodies (Ab), hepatitis B virus antigen (HBsAg) and antihepatitis C virus (HCV) Ab were screened using routine operational testing procedures at the centres. Sensitivity of detection using RDTs was high for HIV Ab-positive samples, but low for intermediately HBsAg (51·5%) and HCV Ab (40·6%)-positive samples. These findings suggest that current RDT use in Africa could pose a hazard to blood safety. PMID:26646317

Seroepidemiologic survey for Chlamydia suis in wild boar (Sus scrofa) populations in Italy.

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Di Francesco, Antonietta; Donati, Manuela; Morandi, Federico; Renzi, Maria; Masia, Marco Antonio; Ostanello, Fabio; Salvatore, Daniela; Cevenini, Roberto; Baldelli, Raffaella

2011-07-01

We used serology to estimate the prevalence of exposure to chlamydiae in Italian populations of wild boars (Sus scrofa). Sera from 173 hunter-killed wild boars harvested during the 2006-2009 hunting seasons in three Italian regions were tested for antibodies to Chlamydia suis, Chlamydophila pecorum, Chlamydophila abortus, and Chlamydophila psittaci by the microimmunofluorescence test. Antibody titers to chlamydiae ≥ 1:32 were detected in 110 of the 173 samples tested (63.6%). Specific reactivity could be assessed only in 44 sera with antibody titers to C. suis that were two- to threefold higher than antibody titers against the other chlamydial species; the other 66 sera had similar reactivity against all the chlamydia species tested. Antibody to C. suis was detected in sera from wild boar populations with rare or no known contact with domestic pigs. These results suggest that the wild boar could be a chlamydia reservoir and may acquire chlamydiae independent of contacts with the domestic pig.

Effects of Different Sera Conditions on Olfactory Ensheathing Cells in Vitro

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Meng Lu

2014-12-01

Full Text Available Transplantation of olfactory ensheathing cells (OEC is a promising therapy in spinal cord injury (SCI treatment. However, the therapeutic efficacy of this method is unstable due to unknown reasons. Considering the alterations in the culture environment that occur during OEC preparation for transplantation, we hypothesize that these changes may cause variations in the curative effects of this method. In this study, we compared OEC cultured in medium containing different types and concentrations of serum. After purification and passage, the OEC were cultured for 7 days in different media containing 5%, 10%, 15% or 20% fetal bovine serum (FBS or rat serum (RS, or the cells were cultured in FBS-containing medium first, followed by medium containing RS. In another group, the OEC were first cultured in 10% FBS for 3 days and then cultured with rat spinal cord explants with 10% RS for another 4 days. An MTT assay and P75 neurotrophin receptor immunofluorescence staining were used to examine cell viability and OEC numbers, respectively. The concentration of neurotrophin-3 (NT-3, which is secreted by OEC into the culture supernatant, was detected using the enzyme-linked immunosorbent assay (ELISA. RT-PCR was applied to investigate the NT-3 gene expression in OEC according to different groups. Compared with FBS, RS reduced OEC proliferation in relation to OEC counts (χ2 = 166.279, df = 1, p < 0.01, the optical density (OD value in the MTT assay (χ2 = 34.730, df = 1, p < 0.01, and NT-3 concentration in the supernatant (χ2 = 242.997, df = 1, p < 0.01. OEC cultured with spinal cord explants secreted less NT-3 than OEC cultured alone (F = 9.611, df = 5.139, p < 0.01. Meanwhile, the order of application of different sera was not influential. There was statistically significant difference in NT-3 gene expression among different groups when the serum concentration was 15% (χ2 = 64.347, df = 1, p < 0.01. In conclusion, different serum conditions may be

Laparoscopy After Previous Laparotomy

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Zulfo Godinjak

2006-11-01

Full Text Available Following the abdominal surgery, extensive adhesions often occur and they can cause difficulties during laparoscopic operations. However, previous laparotomy is not considered to be a contraindication for laparoscopy. The aim of this study is to present that an insertion of Veres needle in the region of umbilicus is a safe method for creating a pneumoperitoneum for laparoscopic operations after previous laparotomy. In the last three years, we have performed 144 laparoscopic operations in patients that previously underwent one or two laparotomies. Pathology of digestive system, genital organs, Cesarean Section or abdominal war injuries were the most common causes of previouslaparotomy. During those operations or during entering into abdominal cavity we have not experienced any complications, while in 7 patients we performed conversion to laparotomy following the diagnostic laparoscopy. In all patients an insertion of Veres needle and trocar insertion in the umbilical region was performed, namely a technique of closed laparoscopy. Not even in one patient adhesions in the region of umbilicus were found, and no abdominal organs were injured.

Investigation of the response to the enterobacterial common antigen after typhoid vaccination

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Arlete M. Milhomem

1987-03-01

Full Text Available Antibodies against the Salmonella typhi enterobacterial common antigen (ECA and the O and H antigens were investigated in sera from healthy male subjects who had been previously vaccinated with the typhoid vaccine. No serological response to ECA was observed. Sera from subjects not previously vaccinated presented titers of ECA hemagglutinins which quantitatively were related to the presence ofH titers, but not to O agglutinins but with no statistical significance. The results are discussed in relation to the possible protective immunological mechanisms in typhoid fever.

Validation of high throughput screening of human sera for detection of anti-PA IgG by Enzyme-Linked Immunosorbent Assay (ELISA) as an emergency response to an anthrax incident

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Semenova, Vera A.; Steward-Clark, Evelene; Maniatis, Panagiotis; Epperson, Monica; Sabnis, Amit; Schiffer, Jarad

2017-01-01

To improve surge testing capability for a response to a release of Bacillus anthracis, the CDC anti-Protective Antigen (PA) IgG Enzyme-Linked Immunosorbent Assay (ELISA) was re-designed into a high throughput screening format. The following assay performance parameters were evaluated: goodness of fit (measured as the mean reference standard r2), accuracy (measured as percent error), precision (measured as coefficient of variance (CV)), lower limit of detection (LLOD), lower limit of quantification (LLOQ), dilutional linearity, diagnostic sensitivity (DSN) and diagnostic specificity (DSP). The paired sets of data for each sample were evaluated by Concordance Correlation Coefficient (CCC) analysis. The goodness of fit was 0.999; percent error between the expected and observed concentration for each sample ranged from −4.6% to 14.4%. The coefficient of variance ranged from 9.0% to 21.2%. The assay LLOQ was 2.6 μg/mL. The regression analysis results for dilutional linearity data were r2 = 0.952, slope = 1.02 and intercept = −0.03. CCC between assays was 0.974 for the median concentration of serum samples. The accuracy and precision components of CCC were 0.997 and 0.977, respectively. This high throughput screening assay is precise, accurate, sensitive and specific. Anti-PA IgG concentrations determined using two different assays proved high levels of agreement. The method will improve surge testing capability 18-fold from 4 to 72 sera per assay plate. PMID:27814939

Development and validation of an epitope-blocking ELISA using an anti-haemagglutinin monoclonal antibody for specific detection of antibodies in sheep and goat sera directed against peste des petits ruminants virus.

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Bodjo, Sanne Charles; Baziki, Jean-de-Dieu; Nwankpa, Nick; Chitsungo, Ethel; Koffi, Yao Mathurin; Couacy-Hymann, Emmanuel; Diop, Mariame; Gizaw, Daniel; Tajelser, Idris Badri Adam; Lelenta, Mamadou; Diallo, Adama; Tounkara, Karim

2018-07-01

Peste des petits ruminants (PPR) is a contagious and economically important disease affecting production of small ruminants (i.e., sheep and goats). Taking into consideration the lessons learnt from the Global Rinderpest Eradication Programme (GREP), PPR is now targeted by the international veterinary community as the next animal disease to be eradicated. To support the African continental programme for the control of PPR, the Pan African Veterinary Vaccine Centre of the African Union (AU-PANVAC) is developing diagnostics tools. Here, we describe the development of a blocking enzyme-linked immunosorbent assay (bELISA) that allows testing of a large number of samples for specific detection of antibodies directed against PPR virus in sheep and goat sera. The PPR bELISA uses an anti-haemagglutinin (H) monoclonal antibody (MAb) as a competitor antibody, and tests results are interpreted using the percentage of inhibition (PI) of MAb binding generated by the serum sample. PI values below or equal to 18% (PI ≤ 18%) are negative, PI values greater than or equal to 25% (PI ≥ 25%) are positive, and PI values greater than 18% and below 25% are doubtful. The diagnostic specificity (DSp) and diagnostic sensitivity (DSe) were found to be 100% and 93.74%, respectively. The H-based PPR-bELISA showed good correlation with the virus neutralization test (VNT), the gold standard test, with a kappa value of 0.947. The H-based PPR-bELISA is more specific than the commercial kit ID Screen® PPR Competition (N-based PPR-cELISA) from IDvet (France), but the commercial kit is slightly more sensitive than the H-based PPR-bELISA. The validation process also indicated good repeatability and reproducibility of the H-based PPR-bELISA, making this new test a suitable tool for the surveillance and sero-monitoring of the vaccination campaign.

Analysis of post-operative changes in serum protein expression profiles from colorectal cancer patients by MALDI-TOF mass spectrometry: a pilot methodological study

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Marsh Simon

2010-04-01

Full Text Available Abstract Background Mass spectrometry-based protein expression profiling of blood sera can be used to discriminate colorectal cancer (CRC patients from unaffected individuals. In a pilot methodological study, we have evaluated the changes in protein expression profiles of sera from CRC patients that occur following surgery to establish the potential of this approach for monitoring post-surgical response and possible early prediction of disease recurrence. Methods In this initial pilot study, serum specimens from 11 cancer patients taken immediately prior to surgery and at approximately 6 weeks following surgery were analysed alongside 10 normal control sera by matrix-assisted laser desorption ionisation time of-flight-mass spectrometry (MALDI-TOF MS. Using a two-sided t-test the top 20 ranked protein peaks that discriminate normal from pre-operative sera were identified. These were used to classify post-operative sera by hierarchical clustering analysis (Spearman's Rank correlation and, as an independent 'test' dataset, by k-nearest neighbour and weighted voting supervised learning algorithms. Results Hierarchical cluster analysis classified post-operative sera from all six early Dukes' stage (A and B patients as normal. The remaining five post-operative sera from more advanced Dukes' stages (C1 and C2 were classified as cancer. Analysis by supervised learning algorithms similarly grouped all advanced Dukes' stages as cancer, with four of the six post-operative sera from early Dukes' stages being classified as normal (P = 0.045; Fisher's exact test. Conclusions The results of this pilot methodological study illustrate the proof-of-concept of using protein expression profiling of post-surgical blood sera from individual patients to monitor disease course. Further validation on a larger patient cohort and using an independent post-operative sera dataset would be required to evaluate the potential clinical relevance of this approach. Prospective

Comparisons of fully automated syphilis tests with conventional VDRL and FTA-ABS tests.

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Choi, Seung Jun; Park, Yongjung; Lee, Eun Young; Kim, Sinyoung; Kim, Hyon-Suk

2013-06-01

Serologic tests are widely used for the diagnosis of syphilis. However, conventional methods require well-trained technicians to produce reliable results. We compared automated nontreponemal and treponemal tests with conventional methods. The HiSens Auto Rapid Plasma Reagin (AutoRPR) and Treponema Pallidum particle agglutination (AutoTPPA) tests, which utilize latex turbidimetric immunoassay, were assessed. A total of 504 sera were assayed by AutoRPR, AutoTPPA, conventional VDRL and FTA-ABS. Among them, 250 samples were also tested by conventional TPPA. The concordance rate between the results of VDRL and AutoRPR was 67.5%, and 164 discrepant cases were all VDRL reactive but AutoRPR negative. In the 164 cases, 133 showed FTA-ABS reactivity. Medical records of 106 among the 133 cases were reviewed, and 82 among 106 specimens were found to be collected from patients already treated for syphilis. The concordance rate between the results of AutoTPPA and FTA-ABS was 97.8%. The results of conventional TPPA and AutoTPPA for 250 samples were concordant in 241 cases (96.4%). AutoRPR showed higher specificity than that of VDRL, while VDRL demonstrated higher sensitivity than that of AutoRPR regardless of whether the patients had been already treated for syphilis or not. Both FTA-ABS and AutoTPPA showed high sensitivities and specificities greater than 98.0%. Automated RPR and TPPA tests could be alternatives to conventional syphilis tests, and AutoRPR would be particularly suitable in treatment monitoring, since results by AutoRPR in cases after treatment became negative more rapidly than by VDRL. Copyright © 2013. Published by Elsevier Inc.

Performance study of the automated immunoassay test anti-hepatitis C virus VIDAS® for the qualitative detection of antibodies anti-hepatitis C virus

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Sara Salvetti

2016-03-01

Full Text Available Background and aims: Hepatitis C virus (HCV represents a major worldwide public health problem requiring global action for the diagnosis, treatment and prevention of this infection. HCV is the leading cause of chronic liver disease, including chronic hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma and it is responsible for about 350,000 deaths yearly. Anti-HCV assays remain the first choice for screening HCV infection in most clinical laboratories. The anti-HCV VIDAS® (bioMérieux, Marcy L’Etoile, France test has been recently launched and we aimed to evaluate its performance compared with other widely used methods. Materials and methods: Routine anti-HCV screening of clinical samples was carried out on the ARCHITECT® i2000SR platform (Abbot Laboratories, Abbott Park, IL, USA; out of 8876 samples tested from October 2012 to May 2013, 70 sera with low positive anti-HCV results (1≤S/CO<8 were collected. These samples were tested for the presence of anti-HCV antibodies using anti-HCV VIDAS® and INNO-LIATM HCV score assays (Innogenetics NV, Ghent, Belgium. Results and conclusions: Positive anti-HCV results were obtained in 61.4% and 41.4% of sera tested with VIDAS® and INNO-LIATM, respectively. Concordance between methods was 63.2% for ARCHITECT® and VIDAS®, 42.6% for ARCHITECT® and INNO-LIATM, and 79.4% for VIDAS® and INNO-LIATM. Anti-HCV VIDAS® demonstrated superior specificity compared to the anti-HCV test ARCHITECT®; therefore, this assay has been introduced in our routine analysis as a second level screening test to select samples to be subjected to confirmatory anti- HCV immunoblot.

Brucella melitensis VirB12 recombinant protein is a potential marker for serodiagnosis of human brucellosis.

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Mirkalantari, Shiva; Zarnani, Amir-Hassan; Nazari, Mahboobeh; Irajian, Gholam Reza; Amirmozafari, Nour

2017-03-03

The numerous drawbacks of current serological tests for diagnosis of brucellosis which mainly results from cross reactivity with LPS from other gram-negative bacteria have generated an increasing interest to find more specific non-LPS antigens. Previous studies had indicated that Brucella VirB12 protein, a cell surface protein and component of type IV secretion system, induces antibody response during animal infection. However, this protein has not yet been tested as a serological diagnostic marker in human brucellosis. Recombinant VirB12 protein was prepared and evaluated the efficacy of it in an indirect enzyme-linked immunosorbent assay (ELISA) for brucellosis with sera collected from different region of Iran and the results were compared with a commercial ELISA kit. Sera from human brucellosis patients strongly reacted to the purified recombinant VirB12. The sensitivity, specificity, accuracy, negative predictive value and positive predictive value of recombinant VirB12-based ELISA related to the commercial-ELISA method were 87.8, 94, 90, 80 and 96.6% respectively. We concluded that antigenic VirB12 have a property value that can be considered as a candidate for using in serodiagnostic tests for human brucellosis.

Retrospective study of pestivirus infection in Pyrenean chamois (Rupicapra pyrenaica) and other ungulates in the Pyrenees (NE Spain).

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Marco, Ignasi; Cabezón, Oscar; Rosell, Rosa; Fernández-Sirera, Laura; Allepuz, Alberto; Lavín, Santiago

2011-04-21

In 2001 a new Pestivirus (Family Flaviviridae) was associated with an outbreak of a previously unreported disease in Pyrenean chamois (Rupicapra pyrenaica) in the Pyrenees (NE Spain). Molecular characterization assigned this virus to the Border Disease Virus (BDV) cluster, BDV-4 genotype. A retrospective study was performed in archived sera and spleen of 74 Pyrenean chamois and in archived sera of 28 mouflon (Ovis ammon), 56 red deer (Cervus elaphus), 43 roe deer (Capreolus capreolus) and 29 fallow deer (Dama dama) from the Pyrenees between the years 1990 and 2000. Thirty six of 74 (48.6%) sera of Pyrenean chamois, one of mouflon and one of red deer were positive by an ELISA antibody test. Comparative virus neutralization tests were performed on 26 seropositive chamois, one mouflon and one red deer, using five pestivirus strains. An ELISA antigen test was performed on 37 seronegative chamois and yielded positive results in one chamois and inconclusive result in two. RT-PCR and virus isolation performed on spleen samples from these three animals gave positive results in the positive and one inconclusive animal. Sequence analysis in the 5' unstranslated region revealed that they were grouped into the BDV-4 genotype. Virological and serological data of the present study indicate that BDV infection has been present in the chamois population since at least 1990, 11 years before the first outbreak of disease. Therefore, the emergence of the disease in 2001 is apparently due to other factors rather than the introduction of a new virus in the chamois population. Copyright © 2010 Elsevier B.V. All rights reserved.

Female Sexual Dysfunction in the Late Postpartum Period Among Women with Previous Gestational Diabetes Mellitus

International Nuclear Information System (INIS)

Sargin, M. A.; Yassa, M.; Taymur, B. D.; Akca, G.; Tug, N.; Taymur, B.

2017-01-01

Objective: To compare the status of female sexual dysfunction (FSD) between women with a history of previous gestational diabetes mellitus (GDM) and those with follow-up of a healthy pregnancy, using the female sexual function index (FSFI) questionnaire. Study Design: Cross-sectional study. Place and Duration of Study: Department of Obstetrics and Gynecology, Fatih Sultan Mehmet Training and Research Hospital, Istanbul, Turkey, from September to December 2015. Methodology: Healthy sexually active adult parous females were included. Participants were asked to complete the validated Turkish versions of the FSFI and Hospital Anxiety and Depression Scale (HADS) questionnaires. Student's t-test was used for two-group comparisons of normally distributed variables and quantitative data. Mann-Whitney U-test was used for two-group comparisons of non-normally distributed variables. Pearson's chi-squared test, the Fisher-Freeman-Halton test, Fisher's exact test, and Yates' continuity correction test were used for comparison of qualitative data. Results: The mean FSFI scores of the 179 participants was 23.50 +- 3.94. FSFI scores and scores of desire, arousal, lubrication, orgasm, satisfaction, and pain were not statistically significantly different (p>0.05), according to a history of GDM and types of FSD (none, mild, severe). HADS scores and anxiety and depression types did not statistically significantly differ according to the history of GDM (p>0.05). Conclusion: An association could not be found in FSFI scores between participants with both the history of previous GDM and with healthy pregnancy; subclinical sexual dysfunction may be observed in the late postpartum period among women with a history of previous GDM. This may adversely affect their sexual health. (author)

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.

Improving Seroreactivity-Based Detection of Glioma

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Nicole Ludwig

2009-12-01

Full Text Available Seroreactivity profiling emerges as valuable technique for minimal invasive cancer detection. Recently, we provided first evidence for the applicability of serum profiling of glioma using a limited number of immunogenic antigens. Here, we screened 57 glioma and 60 healthy sera for autoantibodies against 1827 Escherichia coli expressed clones, including 509 in-frame peptide sequences. By a linear support vector machine approach, we calculated mean specificity, sensitivity, and accuracy of 100 repetitive classifications. We were able to differentiate glioma sera from sera of the healthy controls with a specificity of 90.28%, a sensitivity of 87.31% and an accuracy of 88.84%. We were also able to differentiate World Health Organization grade IV glioma sera from healthy sera with a specificity of 98.45%, a sensitivity of 80.93%, and an accuracy of 92.88%. To rank the antigens according to their information content, we computed the area under the receiver operator characteristic curve value for each clone. Altogether, we found 46 immunogenic clones including 16 in-frame clones that were informative for the classification of glioma sera versus healthy sera. For the separation of glioblastoma versus healthy sera, we found 91 informative clones including 26 in-frame clones. The best-suited in-frame clone for the classification glioma sera versus healthy sera corresponded to the vimentin gene (VIM that was previously associated with glioma. In the future, autoantibody signatures in glioma not only may prove useful for diagnosis but also offer the prospect for a personalized immune-based therapy.

Immunochemical studies of infectious mononucleosis

International Nuclear Information System (INIS)

Fletcher, M.A.; Lo, T.M.; Levey, B.A.; Graves, W.R.

1977-01-01

The coated-tube method of solid-phase radioimmunoassay has been adapted to the detection of heterophile antibodies and antigens of infectious mononucleosis. Disposable plastic hemagglutination trays were coated with purified glycoprotein from horse erythrocytes and the subsequent uptake of antibody from test sera was detected by radio-iodinated horse erythrocyte glycoprotein. In a preliminary survey of sera from patients with infectious mononucleosis and sera from controls, the assay proved highly sensitive and specific. The test system was also useful in a competitive binding assay for immunochemical studies of glycoproteins from other heterophile antigen-positive species

Salivary IgA against sporozoite-specific embryogenesis-related protein (TgERP) in the study of horizontally transmitted toxoplasmosis via T. gondii oocysts in endemic settings

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The prevalence of toxoplasmosis was investigated in endemic settings in Brazil, and calculated by measuring antibodies in two ELISA systems: 1) IgG and IgM from sera tested by commercial conventional ELISA, and 2) IgA, from saliva, and IgG from sera samples tested against a sporozoite-specific prote...

Cloning of a cDNA encoding a surface antigen of Schistosoma mansoni schistosomula recognized by sera of vassinated mice

International Nuclear Information System (INIS)

Dalton, J.P.; Tom, T.D.; Strand, M.

1987-01-01

Spleen cells of mice vaccinated with radiation-attenuated Schistosoma mansoni cercariae were used to produce monoclonal antibodies directed against newly transformed schistosomular surface antigens. One of these monoclonal antibodies recognized a polypeptide of 18 kDa. Binding was measured by radioimmunoassay. This glycoprotein was purified by monoclonal antibody immunoaffinity chromatography and a polyclonal antiserum was prepared against it. Immunofluorescence assays showed that the polyclonal antiserum bound to the surface of newly transformed schistosomula and lung-stage organisms but not to the surface of liver-stage and adult worms. Using this polyclonal antiserum we isolated recombinant clones from an adult worm cDNA expression library constructed in λgt11. Clone 654.2 contained an insert of 0.52 kilobase and hybridized to a 1.2-kilobase mRNA species from adult worms. Most importantly, clone 654.2 produced a fusion protein of 125 kDa that was reactive with sera of vaccinated mice that are capable of transferring resistance. This result encourages future vaccination trials with the fusion protein

The in vitro protection of human decay accelerating factor and hDAF/heme oxygenase-1 transgenes in porcine aortic endothelial cells against sera of Formosan macaques.

Science.gov (United States)

Tu, C-F; Tai, H-C; Wu, C-P; Ho, L-L; Lin, Y-J; Hwang, C-S; Yang, T-S; Lee, J-M; Tseng, Y-L; Huang, C-C; Weng, C-N; Lee, P-H

2010-01-01

To mitigate hyperacute rejection, pigs have been generated with alpha-Gal transferase gene knockout and transgenic expression of human decay accelerating factor (hDAF), MCP, and CD59. Additionally, heme-oxygenase-1 (HO-1) has been suggested to defend endothelial cells. Sera (MS) (0%, 1%, 5%, 10%, and 15%) from Formosan macaques (Macaca cyclopis, MC), an Old World monkey wildly populated in Taiwan, was used to test the protective in vitro, effects of hDAF or hDAF/hHO-1 on porcine aortic endothelial cells (pAEC) derived from hDAF(+), hDAF(+)/hHO-1(+), and hDAF(+)/hHO-1(-) and 1 nontransgenic pAEC. Ten percent human serum (HS) served as a positive control. When MS addition increased to 10% or 15%, all transgenic pAEC exhibited a greater survival than nontransgenic pAEC. Noticeably, 15% MS reduced survived to 40% in nontransgenic and transgenic pAEC, respectively. These results revealed that hDAF exerted protective effects against MC complement activation. However, comparing with 10% MS and HS in pAEC of nontransgenic pigs, the survivability was higher in HS, suggesting that complement activation by MS was more toxic than that by HS. Furthermore, hDAF(+)/hHO-1(+) showed no further protection against effects of MS on transgenic pAEC. Copyright 2010 Elsevier Inc. All rights reserved.

Effect of ingested human antibodies induced by RTS, S/AS01 malaria vaccination in children on Plasmodium falciparum oocyst formation and sporogony in mosquitoes

DEFF Research Database (Denmark)

Miura, Kazutoyo; Jongert, Erik; Deng, Bingbing

2014-01-01

falciparum CS protein, but the ability of serum from vaccinated individuals to inhibit sporogony in mosquitoes has not been evaluated. METHODS: Previously a double-blind, randomized trial of RTS,S/AS01 vaccine, as compared with rabies vaccine, in five- to 17-month old children in Tanzania was conducted....... In this study, polyclonal human antibodies were purified from the pools of sera taken one month after the third vaccination. IgGs were purified from four pools of sera from 25 RTS,S/AS01 vaccinated children each, and two pools of sera from 25 children vaccinated with rabies vaccine each. The ability...

The use of a liquid phase blocking ELISA kit for detection of antibodies against foot-and-mouth disease virus in Colombia

International Nuclear Information System (INIS)

Sanchez Martinez, C.; Quintero, M.

1998-01-01

The objective of this study was to undertake an interlaboratory comparison of a liquid phase blocking ELISA for detection of antibodies to FMD virus. For that purpose sera from 120 vaccinated, 120 infected and 120 FMD negative cattle were tested. All sera were tested in a screening assay at a dilution of 1/32. Positive sera were tested in a titration assay (1/10, 1/50, 1/250, 1/1250). For serotype O 1 Cruzeiro 108 sera from the FMD-free group were classified as negatives giving a specificity of 90%. For the same serotype the group of infected/vaccinated cattle gave 114/115 positive results showing a sensitivity of 95% respectively 96%. For serotype A 24 Cruzeiro from the FMD-free group 85 sera were classified as negatives giving a specificity 71%. For the same serotype the group of infected/vaccinated cattle gave 90/99 positive results showing a sensitivity of 75% respectively 82%. The predictive value of the assay was good as results expected for the different serum categories were mainly confirmed in the test. Nevertheless a high number of plates were rejected due to 'outside limits' and further adjustments are necessary to obtain more reliable results. (author)

Influence of Vitamins on Secondary Reactive Oxygen Species Production in Sera of Patients with Resectable NSCLC

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Thierry Patrice

2016-07-01

Full Text Available Background: Singlet oxygen (1O2 oxidizes targets through the production of secondary reactive oxygen species (SOS. Cancers induce oxidative stress changing with progression, the resulting antioxidant status differing from one patient to the other. The aim of this study was to determine the oxidative status of patients with resectable Non-Small cell lung cancers (NSCLC and the potential influence of antioxidants, compared to sera from healthy donors. Materials and Methods: Serum samples from 10 women and 28 men, 19 adenocarcinomas (ADK, 15 patients N1 or M1 were submitted to a photoreaction producing 1O2. Then, samples were supplemented with vitamins (Vit C, Vit E, or glutathione (GSH. Results: Squamous cell carcinomas (SCC and metastatic SCCs induced a lower SOS rate. While Vit C increased SOS in controls as in patients with metastases, Vit E or the combination of Vit E and C strongly reduced SOS. GSH alone lightly decreased SOS in controls but had no effect in patients either alone or combined with Vit C. Conclusion: In “early” lung cancers, SOS are comparable or lower than for healthy persons. The role of Vitamins varies with gender, cancer type, and metastases. This suggests that an eventual supplementation should be performed on a per-patient basis to evidence any effect.

Antibodies against deamidated gliadin peptides identify adult coeliac disease patients negative for antibodies against endomysium and tissue transglutaminase.

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Dahle, C; Hagman, A; Ignatova, S; Ström, M

2010-07-01

This study was done to evaluate the diagnostic utility of antibodies against deamidated gliadin peptides compared to traditional markers for coeliac disease. To evaluate diagnostic utility of antibodies against deamidated gliadin peptide (DGP). Sera from 176 adults, referred for endoscopy without previous analysis of antibodies against tissue transglutaminase (tTG) or endomysium (EmA), were retrospectively analysed by ELISAs detecting IgA/IgG antibodies against DGP or a mixture of DGP and tTG, and compared with IgA-tTG and EmA. Seventy-nine individuals were diagnosed with coeliac disease. Receiver operating characteristic analyses verified the manufacturers' cut-off limits except for IgA/IgG-DGP/tTG. In sera without IgA deficiency, the sensitivity was higher for IgA/IgG-DGP (0.85-0.87) compared with IgA-tTg (0.76) and EmA (0.61). All tests showed high specificity (0.95-1.00). Eighteen coeliac disease-sera were negative regarding IgA-tTG, nine of which were positive for IgA/IgG-DGP. Sera from coeliac disease-patients >70 years were more often negative for IgA-tTG (50%) and IgA/IgG-DGP (36%) than younger patients (15% and 8% respectively) (P adult coeliac disease patients negative for antibodies against endomysium and tissue transglutaminase. Serology is often negative in elderly patients with coeliac disease; a small bowel biopsy should therefore be performed generously before coeliac disease is excluded.

Serological and microbial survey of Mycoplasma gallisepticum in wild turkeys (Meleagris gallopavo) from six western states.

Science.gov (United States)

Fritz, B A; Thomas, C B; Yuill, T M

1992-01-01

From 1986 to 1989, sera from wild turkeys (Meleagris gallopavo), including three subspecies (M. gallopavo intermedia, M. gallopavo merriami and M. gallopavo mexicana) trapped in six western states were tested for antibody to Mycoplasma gallisepticum (MG) (n = 724), M. synoviae (MS) (n = 461) and M. meleagridis (MM) (n = 354) using the rapid plate agglutination (RPA) assay. Subsamples of these sera were also evaluated using the hemagglutination inhibition (HI) assay for antibody to MG (n = 664) and MS (n = 403). Attempts were made to isolate mycoplasmas by swabbing the trachea and cloaca of 190 live wild turkeys and from various tissues (sinus, nasal turbinates, trachea, lung, ovaries and oviduct) from 76 turkeys at necropsy. Isolates were identified using an immunobinding assay. Seroprevalence of MG, MS and MM in the RPA test was highly variable among years and geographic sites, ranging from 0 to 85%, 0 to 87%, and 0 to 83%, respectively, for each mycoplasma species. Of the 724 wild turkey sera tested, 200 (28%) were positive using the RPA assay, while only 20 (3%) of 664 sera tested using the HI assay were positive (at a titer greater than/= 1:80) for antibody to MG. Of the 461 sera tested 178 (39%) were RPA positive for MS, whereas none of the 403 samples tested by HI were positive for MS. Antibody to MM was detected in 72 (20%) of 354 turkey sera tested by RPA. Mycoplasmas were cultured from 81 (30%) of 266 wild turkeys, including 48 that were sampled live and 33 that were examined by necropsy. Mycoplasmas were isolated from every population in which culture was attempted. M. gallopavonis (MGP) was isolated from 37 (46%) of 81 birds which yielded mycoplasma, representing seven of 12 populations sampled. MG was isolated from lower respiratory tissues of one Rio Grande wild turkey trapped in Texas. M. synoviae was isolated from five of 16 Merriam's wild turkeys trapped in Arizona. Sera of birds from which MG or MS was isolated were positive to the respective

Use of enzyme linked immunosorbent assay (ELISA) for the diagnosis of brucellosis in cattle in Yucatan, Mexico

International Nuclear Information System (INIS)

Dajer, A.; Gutierrez, E.; Zapata, D.

1998-01-01

Sera (247) from non-vaccinated brucellosis negative herds, 328 negative sera from Brucella abortus strain 19 vaccinated herds (brucellosis free), and 95 sera positive to the Rose Bengal test (RBT) and Complement Fixation test (CFT) from Brucella abortus-infected herds, were used to determine the relative sensitivity and specificity of a FAO/IAEA I-ELISA kit and the Rivanol Agglutination Test (RAT), using the CFT as a 'gold standard'. A threshold value for the I-ELISA was determined to be 37 PP using the mean plus 3 standard deviations of the negative sera from vaccinated animals. The I-ELISA showed a high relative sensitivity (100%) and a good relative specificity (92.5%), using the threshold determined for local conditions. The RAT gave a lower sensitivity value than the CFT (97.8%) and good specificity (99.3%). The I-ELISA could be used as a screening test under Yucatan conditions or as a confirmatory test in places where vaccination is not carried out. The RAT lacks sensitivity and is therefore not recommended for use in final stages of eradication programs but could be used in control programmes or early stages of eradication campaigns as a confirmatory test. (author)

Dual infection with hepatitis A and E viruses in outbreaks and in sporadic clinical cases: Cuba 1998-2003.

Science.gov (United States)

Rodríguez Lay, Licel de los Angeles; Quintana, Ariel; Villalba, María Caridad Montalvo; Lemos, Gilda; Corredor, Marité Bello; Moreno, Aidonis Gutiérrez; Prieto, Pablo Aguiar; Guzmán, María G; Anderson, David

2008-05-01

Viral hepatitis ranks as the fifth cause of morbidity for infectious diseases in Cuba. Epidemics are observed frequently in the population, the hepatitis A virus being the main agent responsible for such epidemics. Previous reports also confirmed the circulation of the hepatitis E virus. From 1998 to 2003, 258 serum samples were collected by the Reference Laboratory on Viral Hepatitis during 33 outbreaks of acute viral hepatitis as well as from 39 sporadic clinical cases. Sera were tested for anti-HAV and anti-HEV IgM by EIA. Overall of the 33 outbreaks studied sera from 12 (36.4%) were positive for anti-HAV IgM only, from 7 (21.2%) were positive for anti-HEV IgM only, and from 14 (42.4%) were positive for antibodies to both viruses. Individually of the 258 sera collected, 137 (53.1%) were positives for anti-HAV IgM, 20 (7.8%) were positives for anti-HEV IgM, 33 (12.8%) were positives for both markers and 68 (26.4%) were negative to both. Of the clinical cases, 4 (10.3%) were positives for anti-HAV IgM, 13 (33.3%) were positives for anti-HEV IgM and 5 (12.8%) were positives for both markers. Seventeen (43.6%) sera were negatives for all viral hepatitis markers available (A-E). A high positivity for HEV was found in outbreaks tested with the kit produced by CIGB. In particular HEV seems to infect individuals of all ages. The results demonstrate the co-circulation of and co-infection with two enterically transmitted viruses; however a higher positivity was observed for anti-HAV than to anti-HEV (53.1% vs. 7.8%) in outbreaks.

HIV risk perception and testing behaviours among men having sex with men (MSM reporting potential transmission risks in the previous 12 months from a large online sample of MSM living in Germany

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Ulrich Marcus

2016-10-01

Full Text Available Abstract Background HIV testing and serostatus awareness are essential to implement biomedical strategies (treatment as prevention; oral chemoprophylaxis, and for effective serostatus-based behaviours (HIV serosorting; strategic positioning. The analysis focuses on the associations between reported sexual risks, the perceived risk for HIV infection, and HIV testing behaviour in order to identify the most relevant barriers for HIV test uptake among MSM living in Germany. Methods MSM were recruited to a nationwide anonymous online-survey in 2013 on MSM social networking/dating sites. Questions covered testing behaviours, reasons for testing decisions, and HIV risk perception (5-point scale. Additional questions addressed arguments in favour of home/ home collection testing (HT. Using descriptive statistics and logistic regression we compared men reporting recent HIV testing (RT; previous 12 month with men never tested (NT in a subsample not previously diagnosed with HIV and reporting ≥2 episodes of condomless anal intercourse (CLAI with a non-steady partner of unknown HIV serostatus in the previous 12 months. Results The subsample consisted of 775 RT (13 % of RT and 396 NT (7 % of NT. The number of CLAI episodes in the last 12 months with non-steady partners of unknown HIV status did not differ significantly between the groups, but RT reported significantly higher numbers of partners (>5 AI partners: 65 vs. 44 %. While perceived risks regarding last AI were comparable between the groups, 49vs. 30 % NT were <30 years, lived more often in towns/villages <100,000 residents (60 vs. 39 %, were less out-particularly towards care providers-about being attracted to men (aOR 10.1; 6.9–14.8, more often identified as bisexual (aOR 3.5; 2.5–4.8, and reported lower testing intentions (aOR 0.08; 0.06–0.11. Perceived risks (67 % and routine testing (49 % were the most common testing reasons for RT, while the strong belief not to be infected

Immunodiagnosis of Echinococcus Infections: Confirmatory Testing and Species Differentiation by a New Commercial Western Blot

Science.gov (United States)

Liance, Martine; Janin, Veronique; Bresson-Hadni, Solange; Vuitton, Dominique-Angele; Houin, Rene; Piarroux, Renaud

2000-01-01

The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis-infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species-specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID:11015390

Evaluation of ELISA and haemagglutination inhibition as screening tests in serosurveillance for H5/H7 avian influenza in commercial chicken flocks

DEFF Research Database (Denmark)

Arnold, M E; Slomka, M J; Breed, A C

2018-01-01

with an ELISA, which detects antibodies to all subtypes. Sera (n = 1055) from 74 commercial chicken flocks were tested by both methods. A Bayesian approach served to estimate diagnostic test sensitivities and specificities, without assuming any 'gold standard'. Sensitivity and specificity of the ELISA was 97......% and 99.8%, and for H5/H7 HI 43% and 99.8%, respectively, although H5/H7 HI sensitivity varied considerably between infected flocks. ELISA therefore provides superior sensitivity for the screening of chicken flocks as part of an algorithm, which subsequently utilises H5/H7 HI to identify infection...

Sandwich immunoassay for alpha-fetoprotein in human sera using gold nanoparticle and magnetic bead labels along with resonance Rayleigh scattering readout

International Nuclear Information System (INIS)

Lu, Yao; Huang, Xiangyi; Ren, Jicun

2013-01-01

We describe a sensitive sandwich immunoassay for alpha-fetoprotein (AFP). It is making use of gold nanoparticles (GNPs) and magnetic beads (MBs) as labels, and of resonance Rayleigh scattering for detection. Two antibodies were labeled with GNPs and MBs, respectively, and MB-antigen-GNP complexes were formed in the presence of antigens. The MB labels also serve as solid phase carriers that can be used to magnetically separate the immuno complex. The GNP labels are used as optical probes, and Rayleigh scattering was used to determine the concentration of free GNPs-antibody after separation of the MB-antigen-GNP complexes. The concentration of AFP is related to the intensity of light scattered by free GNPs in the 13.6 pM to 436 pM concentration range, and the limit of detection is 13.6 pM. The method was applied to the determination of AFP in sera of cancer patients, and the results agree well with those obtained by conventional ELISA. (author)

Tumor Necrosis Factors, Interferons and Matrix Metalloproteinase-9 in Sera of Non-Hodgkin's Lymphoma Patients

International Nuclear Information System (INIS)

Abdel Malak, C.A.; Karawya, E.M.; Hammouda, G.A.; Zakhary, N.I.

2003-01-01

In the present study, the serum levels of some cytokines and the matrix metalloproteinase-9 (MMP-9) were studied in an attempt to find suitable markers for early diagnosis of non- Hodgkin's lymphoma (NHL) and to assess their role in differentiating between disseminated and non disseminated cases. The present study was conducted on 60 patients with non disseminated NHL, 14 patients with disseminated NHL, in addition to 10 healthy controls. Their sera were used to determine tumor necrosis factor-α (TNF--α), tumor necrosis factor--β (TNF-β), interferon---α), (IFN--α), interferon-γ (IFN--γ) and Matrix Metalloproteinase-9 (MMP-9) using the ELISA technique. The results showed that the serum level of TNF---α), and IFN---α), can be used to differentiate between the control group and the group of NHL patients. However, they could not differentiate between non disseminated NHL (nd- NHL) and disseminated NHL (d- NHL). On the other hand, the serum level of TNF-β) can be used to differentiate between nd- NHL and d- NHL, but not between the control group and nd-NHL. Each of [FN--γ and MMP-9 were not useful in discrimination between the control group and the diseased ones. Our data revealed no correlation between serum level of the parameters investigated and the gender of the patients. The present results revealed that TNF-α) and INF-α), can be used as diagnostic tools for NHL. On the other hand, TNF-β) is useful in the differentiation between nd-NHL and d-NHL

Validation of high throughput screening of human sera for detection of anti-PA IgG by Enzyme-Linked Immunosorbent Assay (ELISA) as an emergency response to an anthrax incident.

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Semenova, Vera A; Steward-Clark, Evelene; Maniatis, Panagiotis; Epperson, Monica; Sabnis, Amit; Schiffer, Jarad

2017-01-01

To improve surge testing capability for a response to a release of Bacillus anthracis, the CDC anti-Protective Antigen (PA) IgG Enzyme-Linked Immunosorbent Assay (ELISA) was re-designed into a high throughput screening format. The following assay performance parameters were evaluated: goodness of fit (measured as the mean reference standard r 2 ), accuracy (measured as percent error), precision (measured as coefficient of variance (CV)), lower limit of detection (LLOD), lower limit of quantification (LLOQ), dilutional linearity, diagnostic sensitivity (DSN) and diagnostic specificity (DSP). The paired sets of data for each sample were evaluated by Concordance Correlation Coefficient (CCC) analysis. The goodness of fit was 0.999; percent error between the expected and observed concentration for each sample ranged from -4.6% to 14.4%. The coefficient of variance ranged from 9.0% to 21.2%. The assay LLOQ was 2.6 μg/mL. The regression analysis results for dilutional linearity data were r 2  = 0.952, slope = 1.02 and intercept = -0.03. CCC between assays was 0.974 for the median concentration of serum samples. The accuracy and precision components of CCC were 0.997 and 0.977, respectively. This high throughput screening assay is precise, accurate, sensitive and specific. Anti-PA IgG concentrations determined using two different assays proved high levels of agreement. The method will improve surge testing capability 18-fold from 4 to 72 sera per assay plate. Published by Elsevier Ltd.

[Seroprevalence of tularemia in risk groups of humans and animals in Van, eastern Turkey].

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Bayram, Yasemin; Özkaçmaz, Ayşe; Parlak, Mehmet; Başbuğan, Yıldıray; Kılıç, Selçuk; Güdücüoğlu, Hüseyin

2015-10-01

Tularemia has become a re-emerging zoonotic disease in Turkey recently. The aims of this study were to determine the seroprevalence of tularemia in humans and their animals living in rural risky areas of our region and to investigate the risk factors. Between January and July 2012, people living in rural areas of Van province (located at eastern part of Turkey) and their domestic animals were included in the study. The sample size was determined by using cluster sampling method like in an event with known prevalence and planned as a cross-sectional epidemiological study. Proportional random sampling method was used to determine which individuals will be included in the study. Presence of tularemia antibodies in the sera of a total 495 voluntary persons (343 female, 152 male; age range: 18-79 years, mean age: 40.61) and their 171 animals (40 cattle, 124 sheep and 7 goats) were screened by microagglutination test using safranin O-stained F.tularensis antigen (Public Health Agency of Turkey). For the evaluation of cross-reactivity between Brucella spp., tularemia positive serum samples were also tested with brucella microagglutination test. Among human and animal samples, 11.9% (59/495) and 44% (76/171) yielded positive results with the titers of ≥ 1:20 in F.tularensis microagglutination test, respectively. However, 69.5% (41/59) of human sera and 78.9% (60/76) of animal sera demonstrated equal or higher titers in the brucella test, so those sera were considered as cross-reactive. After exclusion of these sera, the seroprevalence for F.tularensis were calculated as 3.6% (18/495) for humans and 9.4% (16/171) for animals. Among the 16 animals with positive results, 12 were sheep, three were cattle and one was goat. The difference between seropositivity rates among the domestic animal species was not statistically significant (p> 0.05). In addition, no statistically significant differences were found between risk factors including insect bite, tick bite, contact with

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

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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.

High levels of inactive thymidine kinase 1 polypeptide detected in sera from dogs with solid tumours by immunoaffinity methods: implications for in vitro diagnostics.

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Kiran Kumar, J; Sharif, H; Westberg, S; von Euler, H; Eriksson, S

2013-09-01

Determination of serum thymidine kinase 1 (STK1) activity has been used as a proliferation marker for neoplastic diseases in both human and veterinary medicine. The purpose of this study was to determine STK1 activity and enzyme levels in different dog tumours. Serum samples from three dogs with leukaemia, five with lymphoma, 21 with solid tumours and 18 healthy dogs were analyzed for STK1 activity, using an optimized [(3)H]-deoxythymidine (dThd) phosphorylation assay, and for STK1 protein levels using an immunoaffinity/western blot assay. STK1 activity in dogs with haematological tumours was significantly higher than in the solid tumour and healthy dog groups (mean ± standard deviation [SD] = 65 ± 79, 1.1 ± 0.5, and 1.0 ± 0.4 pmol/min/mL, respectively). Serum samples were analyzed after immunoaffinity isolation by western blot and the TK1 26 kDa band intensities quantified revealing that concentrations were significantly higher in dogs with haematological tumours and solid tumours compared to healthy dogs (mean ± SD=33 ± 12, 30 ± 13, and 10 ± 5 ng/mL, respectively). Pre-incubation with the reducing agent dithioerythritol (DTE) showed a decrease in STK1 activity and protein levels in most samples, but an increase of about 20% in sera from healthy dogs and from those with haematological malignancies. Compared to animals with solid tumours, the specific STK1 activity (nmol [(3)H]-deoxythymidine monophosphate (dTMP)/min/mg of TK1 protein of 26 kDa) was 30-fold higher in haematological malignancies and 2.5-fold higher in healthy dogs, respectively. The results demonstrate that there is a large fraction of inactive TK1 protein, particularly in sera from dogs with solid tumours. The findings are important in the use of STK1 as a biomarker. Copyright © 2013 Elsevier Ltd. All rights reserved.

Development and evaluation of an immunochromatographic strip test based on the recombinant UL51 protein for detecting antibody against duck enteritis virus

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Sun Tao

2010-10-01

Full Text Available Abstract Background Duck enteritis virus (DEV infection causes substantial economic losses to the worldwide duck-producing areas. The monitoring of DEV-specific antibodies is a key to evaluate the effect of DEV vaccine and develop rational immunization programs. Thus, in this study, an immunochromatographic strip (ICS test was developed for detecting DEV serum antibodies. Results The ICS test is based on membrane chromatography, and uses both the purified recombinant UL51 protein conjugated with colloidal gold and goat anti-rabbit IgG conjugated with colloidal gold as tracers, the purified recombinant UL51 protein as the capture reagent at the test line, and rabbit IgG as the capture reagent at the control line. The specificity of the ICS was evaluated by sera against DEV, Duck hepatitis virus (DHV, Riemerella anatipestifer (RA, Duck E. coli, Muscovy duck parvovirus (MPV, or Duck Influenza viruses (DIV. Only sera against DEV showed the strong positive results. In order to determine the sensitivity of the ICS, anti-DEV serum diluted serially was tested, and the minimum detection limit of 1:128 was obtained. The ICS components, which are provided in a sealed package, require no refrigeration and are stable for 12 months. To evaluate the effect of the ICS, 110 duck serum samples collected from several non-immune duck flocks were simultaneously tested by the ICS test, enzyme-linked immunosorbent assay (ELISA and neutralization test (NT. The results showed that the sensitivity of the ICS test was almost consistent with ELISA and much higher than NT, has low cost, and is rapid (15 min and easy to perform with no requirement of specialized equipment, reagent or technicians. Conclusions In this work, we successfully developed a simple and rapid ICS test for detecting DEV serum antibodies for the first time. The ICS test was high specific and sensitive for the rapid detection of anti-DEV antibodies, and has great potential to be used for the serological

Validation of a serum neutralization test for detection of antibodies specific to cyprinid herpesvirus 3 in infected common and koi carp (Cyprinus carpio)

DEFF Research Database (Denmark)

Cabon, J.; Louboutin, L.; Castric, J.

2017-01-01

healthy and naturally or experimentally CyHV-3-infected carp. French CyHV-3 isolate 07/108b was neutralized efficiently by sera from carp infected with European, American and Taiwanese CyHV-3 isolates, but no neutralization was observed using sera specific to other aquatic herpesviruses. Diagnostic...

Serological Evidence of Immune Priming by Group A Streptococci in Patients with Acute Rheumatic Fever

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Jeremy M Raynes

2016-07-01

Full Text Available Acute rheumatic fever (ARF is an autoimmune response to Group A Streptococcus (GAS infection. Repeated GAS exposures are proposed to ‘prime’ the immune system for autoimmunity. This notion of immune-priming by multiple GAS infections was first postulated in the 1960s, but direct experimental evidence to support the hypothesis has been lacking. Here we present novel methodology, based on antibody responses to GAS T‑antigens, that enables previous GAS exposures to be mapped in patient sera. T-antigens are surface expressed, type specific antigens and GAS strains fall into 18 major clades or T-types. A panel of recombinant T-antigens was generated and immunoassays were performed in parallel with serum depletion experiments allowing type-specific T‑antigen antibodies to be distinguished from cross-reactive antibodies. At least two distinct GAS exposures were detected in each of the ARF sera tested. Furthermore, no two sera had the same T-antigen reactivity profile suggesting that each patient was exposed to a unique series of GAS T‑types prior to developing ARF. The methods have provided much-needed experimental evidence to substantiate the immune-priming hypothesis, and will facilitate further serological profiling studies that explore the multifaceted interactions between GAS and the host.

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.

Antibodies to elastin peptides in sera of Belgian Draught horses with chronic progressive lymphoedema.

Science.gov (United States)

van Brantegem, L; de Cock, H E V; Affolter, V K; Duchateau, L; Hoogewijs, M K; Govaere, J; Ferraro, G L; Ducatelle, R

2007-09-01

Chronic progressive lymphoedema (CPL) is a recently recognised disease of the lymphatic system characterised by lesions in the skin of the lower legs in several draught horse breeds, including the Belgian Draught hourse. Clinical signs slowly progress and result in severe disfigurement of the limbs. Ideally, supportive treatment should be started early in the disease process. However early diagnosis and monitoring progression of CPL is still a challenge. Elastin changes, characterised by morphological alterations as well as increased desmosine levels, in the skin of the distal limbs of horses affected with CPL are probably associated with a marked release of elastin degradation products, which elicit production of circulating anti-elastin antibodies (AEAbs) in the serum. An enzyme-linked immunosorbent assay (ELISA) for detection of serum AEAbs may document elastin breakdown. An ELISA technique was used to evaluate levels of AEAbs in sera of 97 affected Belgian Draught horses that were clinically healthy except for possible skin lesions, associated with CPL in their distal limbs. The horses were divided into 5 groups according to the severity of these skin lesions: normal horses (Group 1, n = 36), horses with mild lesions (Group 2, n = 43), horses with moderate lesions (Group 3, n = 8), horses with severe lesions (Group 4, n = 10) and, as a control, healthy Warmblood horses, unaffected by the disease (Group 5, n = 83). Horses with clinical signs of CPL had significantly higher AEAb levels compared to clinically normal Belgian Draught horses and to healthy Warmblood horses. These levels correlated with severity of lesions. CPL in draught horses is associated with an increase of serum AEAbs. Evaluation of serum levels of AEAbs by ELISA might be a useful diagnostic aid for CPL. Pathological degradation of elastic fibres, resulting in deficient support of the distal lymphatics, is proposed as a contributing factor for CPL in Belgian Draught horses.

eCD4-Ig promotes ADCC activity of sera from HIV-1-infected patients.

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Meredith E Davis-Gardner

2017-12-01

Full Text Available Antibody-dependent cell-mediated cytotoxity (ADCC can eliminate HIV-1 infected cells, and may help reduce the reservoir of latent virus in infected patients. Sera of HIV-1 positive individuals include a number of antibodies that recognize epitopes usually occluded on HIV-1 envelope glycoprotein (Env trimers. We have recently described eCD4-Ig, a potent and exceptionally broad inhibitor of HIV-1 entry that can be used to protect rhesus macaques from multiple high-dose challenges with simian-human immunodeficiency virus AD8 (SHIV-AD8. Here we show that eCD4-Ig bearing an IgG1 Fc domain (eCD4-IgG1 can mediate efficient ADCC activity against HIV-1 isolates with differing tropisms, and that it does so at least 10-fold more efficiently than CD4-Ig, even when more CD4-Ig molecules bound cell surface-expressed Env. An ADCC-inactive IgG2 form of eCD4-Ig (eCD4-IgG2 exposes V3-loop and CD4-induced epitopes on cell-expressed trimers, and renders HIV-1-infected cells susceptible to ADCC mediated by antibodies of these classes. Moreover, eCD4-IgG2, but not IgG2 forms of the broadly neutralizing antibodies VRC01 and 10-1074, enhances the ADCC activities of serum antibodies from patients by 100-fold, and significantly enhanced killing of two latently infected T-cell lines reactivated by vorinostat or TNFα. Thus eCD4-Ig is qualitatively different from CD4-Ig or neutralizing antibodies in its ability to mediate ADCC, and it may be uniquely useful in treating HIV-1 infection or reducing the reservoir of latently infected cells.

A liquid phase blocking ELISA for the detection of antibodies against infectious bronchitis virus

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Cardoso T.C.

1999-01-01

Full Text Available A liquid phase blocking ELISA (LPB-ELISA was developed for the detection and measurement of antibodies against infectious bronchitis virus (IBV. The purified and nonpurified virus used as antigen, the capture and detector antibodies, and the chicken hyperimmune sera were prepared and standardized for this purpose. A total of 156 sera from vaccinated and 100 from specific pathogen-free chickens with no recorded contact with the virus were tested. The respective serum titers obtained in the serum neutralization test (SNT were compared with those obtained in the LPB-ELISA. There was a high correlation (r2 = 0.8926 between the two tests. The LPB-ELISA represents a single test suitable for the rapid detection of antibodies against bronchitis virus in chicken sera, with good sensitivity (88%, specificity (100% and agreement (95.31%.

Teste de ELISA indireto para o diagnóstico sorológico de pitiose Indirect ELISA for the serodiagnostic of pythiosis

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Janio M. Santurio

2006-03-01

Full Text Available A pitiose, doença granulomatosa de eqüinos causada pelo oomiceto Pythium insidiosum, tem como característica a evolução rápida seguida de morte dos animais. Estas mortes muitas vezes são causadas por diagnósticos errôneos ou demorados quando os doentes já não respondem ao tratamento. Este trabalho teve por objetivo a padronização do ensaio imunoenzimático indireto (ELISA para diagnóstico sorológico de pitiose em eqüinos e coelhos, visando a diminuição de erros e de tempo necessário para o diagnóstico. Para o desenvolvimento e validação do teste foram utilizadas 72 amostras de soro de eqüinos saudáveis e 44 soros de eqüinos com pitiose confirmada. Os resultados da validação do ELISA para eqüinos foram: sensibilidade 97,72%, especificidade 90,27%, valor preditivo positivo 86%, valor preditivo negativo 98,4% e eficiência de 93,1%. Para coelhos, o teste foi padronizado com 48 amostras de soro de animais saudáveis e 24 amostras de coelhos imunizados com antígenos de P. insidiosum. Os resultados foram: sensibilidade 91,66%, especificidade 95,83%, valor preditivo positivo 91,66%, valor preditivo negativo 95,83% e eficiência de 94,44%. Os resultados deste trabalho demonstram que o ensaio imunoenzimático indireto é um método seguro e eficaz para o diagnóstico sorológico da pitiose.Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum that affects humans and animals, especially horses. Deaths are very often the consequence of incorrect or late diagnosis when animals no longer respond to treatment. This study aimed standardization of the ELISA assay for the serodiagnostic of pythiosis in horses and rabbits, in order to minimize errors and delays in the diagnosis of the disease. Sera of 72 healthy and 44 of by pythiosis affected horses were used for development and evaluation of the test. The ELISA for equine diagnostic showed 97.72% sensitivity, 90.27% specificity, 86% positive predictive value

Immune response to a mammary adenocarcinoma. V. Sera from tumor-bearing rats contain multiple factors blocking cell-mediated cytotoxicity.

Science.gov (United States)

Huber, S A; Lucas, Z J

1978-12-01

Sera from Fischer rats 3 to 13 days after i.p. injection of syngeneic 13762A mammary adenocarcinoma contain three factors specifically blocking cell-mediated cytotoxicity (CMC). The major blocking factor is a 160,000-dalton IgG that combines specifically to cytolytic lymphocytes but not to tumor cells or tumor antigen, and that is not dissociated after treatment with 8 M urea. The other factors have been putatively identified as tumor antigen (less than 70,000 daltons) and as soluble antigen-antibody complexes (greater than 200,000 daltons). Injecting the tumor antigen into tumor-free rats induced spleen cells specifically cytotoxic to the 13762A tumor and provided partial protection to challenge with live tumor cells. Treating soluble antigen-antibody complexes with 8 M urea decreased the size of the blocking activity from greater than 200,000 to less than 70,000 daltons. Although the IgG fraction dissociated from the complex did not block CMC, it did recombine with the tumor antigen fraction to transfer activity to the greater than 200,000-dalton fraction. In contrast, mixing tumor antigen with the IgG fraction that did block CMC did not alter the size of the blocking activities.

Prevalence of antibody titers to leptospira spp. in Minnesota white-tailed deer

Science.gov (United States)

Goyal, S.M.; Mech, L.D.; Nelson, M.E.

1992-01-01

Serum samples (n = 204) from 124 white-tailed deer (Odocoileus virginianus) in northeastern Minnesota (USA) were collected from 1984 through 1989 and tested for antibodies to six serovars of Leptospira interrogans (bratislava, canicola, grippotyphosa, hardjo, icterohemorrhagiae, and pomona) using a microtiter agglutination test. Eighty-eight (43%) sera were positive at greater than or equal to 1:100 for antibodies against serovars pomona and/or bratislava; none was positive for any of the other four serovars. None of the 31 sera collected in 1984-85 was positive, whereas all 54 sera collected from 1986 through 1988 had titers of greater than or equal to 1:100. During 1989, only 34 (29%) of 119 sera had titers of greater than or equal to 1:100. Based on these results, we believe there to be wide variability in exposure of Minnesota deer to Leptospira interrogans.

Screening test for neutralizing antibodies against yellow fever virus, based on a flavivirus pseudotype.

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Séverine Mercier-Delarue

Full Text Available Given the possibility of yellow fever virus reintroduction in epidemiologically receptive geographic areas, the risk of vaccine supply disruption is a serious issue. New strategies to reduce the doses of injected vaccines should be evaluated very carefully in terms of immunogenicity. The plaque reduction test for the determination of neutralizing antibodies (PRNT is particularly time-consuming and requires the use of a confinement laboratory. We have developed a new test based on the use of a non-infectious pseudovirus (WN/YF17D. The presence of a reporter gene allows sensitive determination of neutralizing antibodies by flow cytometry. This WN/YF17D test was as sensitive as PRNT for the follow-up of yellow fever vaccinees. Both tests lacked specificity with sera from patients hospitalized for acute Dengue virus infection. Conversely, both assays were strictly negative in adults never exposed to flavivirus infection or vaccination, and in patients sampled some time after acute Dengue infection. This WN/YF17D test will be particularly useful for large epidemiological studies and for screening for neutralizing antibodies against yellow fever virus.

Reactividad inmunoquímica de sueros anti- Caiman yacare y Caiman latirostris frente a sueros de diferentes especies

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de Roodt, Adolfo Rafael

2010-06-01

Full Text Available Se estudió la reactividad inmunoquímica entre los sueros de distintas especies de reptiles frente a sueros hiperinmunes experimentales anti-suero de Caiman yacare y anti-suero de Caiman latirostris. Los sueros que se probaron fueron los homólogos de Caiman yacare, Caiman latirostris y los heterólogos de Alligator missisipiensis, Tupinambis merinae, Tupinambis rufescens, Chelonoidis chilensis, Clelia rustica, Waglerophis merremii, Lystrophys dorbignyi, Phyton molurus, Boa constrictor occidentalis, Eunectes notaeus, Crotalus durissus terrificus, Bothrops alternatus, Bothrops diporus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Pitangus sulphuratus y Gallus gallus. La reactividad inmunoquímica se determinó mediante las técnicas de doble inmunodifusión y ELISA, mostrándose importante entre los sueros de los crocodrílidos y baja entre estos y los de las otras especies de reptiles estudiadas. Se observó mayor reactividad entre los antisueros anti-Caiman respecto a los sueros de Caiman latirostris y Caiman yacare que frente al suero de Alligator missisipiensis. Además, se encontró una fuerte reactividad entre ambos sueros anti-Caiman y el de Gallus gallus poniendo en evidencia la fuerte reactividad entre los sueros de arcosaurios. In order to study the immunochemical reactivity among sera from different species of reptiles regarding sera from Caiman, the immunoreactivity of sera from reptiles against antisera to Caiman yacare or anti-Caiman latirostris sera was studied. These hiperimmune sera were tested against sera from Alligator missisipiensis, Tupinambis merinae, Tupinambis rufescens, Chelonoidis chilensis, Clelia rustica, Waglerophis merremii, Lystrophys dorbignyi, Phyton molurus, Boa constrictor occidentalis, Eunectes notaeus, Crotalus durissus terrificus, Bothrops alternatus, Bothrops neuwiedii, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Pitangus sulphuratus and Gallus gallus. The immunochemical

Patch tests in children: a review of 13 years of experience in comparison with previous data.

Science.gov (United States)

Milingou, Maria; Tagka, Anna; Armenaka, Melina; Kimpouri, Konstantina; Kouimintzis, Dimitris; Katsarou, Alexandra

2010-01-01

The true prevalence of allergic contact dermatitis (ACD) in children remains unknown. Our aim was to compare the results of patch tests in children with suspected ACD between two different periods of time and identify possible changes in emerging allergens. We compared contact allergens, gender, age distribution, and personal history of atopic dermatitis (AD), in correlation with the positivity of patch tests, between two equal periods of time (232 children tested during 1980-1993, period A, and 255 children during 1994-2007, period B) in the same region and in the same institution. Patch test positivity was 47.8% in period A, and 60% in period B (p = 0.083). The most common allergens in period A were: nickel sulfate (16.3%), cobalt chloride (8.6%), fragrance mix (7.3%), potassium dichromate (4.3%), and thimerosal only (1.7%). In period B, the allergen distribution was as follows: nickel sulfate (21.56%), thimerosal (18.03%), cobalt chloride (12.9%), potassium dichromate (9.4%), and fragrance mix (4.7%). Girls were more likely to have a positive patch test compared with boys, with reactions in 53% of girls and 39% of boys in period A (p = 0.003), and 61% of girls and 58% of boys in period B (p = 0.691). Twenty-nine per cent of patients with positive results had a personal history of AD in period A and 44% in period B (p = 0.015). Differences in the positivity of allergens between different time periods reflect changes in habits, of allergens exposure or preventive measures.

A Study for Brucellosis Seroprevelance in Agri

Directory of Open Access Journals (Sweden)

Duran Tok

2009-12-01

Full Text Available AIM: We evaluated retrospectively laboratory test results of 520 patient who has brucellosis suspect between 2002-2004 years. METHOD: We use to Rose-Bengal test, Wright agglutination test and the other laboratory results and demographic properties for diagnosis. RESULTS: Rose-Bengal test was positive in 39 patients (11.3 % sera. Wright agglutination test was found positive for 1/160 or higher titers in 18 (3.4% sera. CONCLUSION: Wright agglutination test gave higher positive results in summer and autumn months. [TAF Prev Med Bull 2009; 8(6.000: 485-488

Validation of basophil histamine release against the autologous serum skin test and outcome of serum-induced basophil histamine release studies in a large population of chronic urticaria patients

DEFF Research Database (Denmark)

Platzer, M H; Grattan, C E H; Poulsen, Lars K.

2005-01-01

the immunoglobulin E (IgE) or the high affinity IgE receptor (FcepsilonRI) and serum-induced histamine release (HR) from basophils and mast cells. We have examined the correlation between the ASST and a new basophil histamine-releasing assay (the HR-Urtikaria test) in a group of well-characterized CU patients...... and subsequently determined the frequency of HR-Urticaria-positive sera from a larger population of CU patients....

Comparative Test Case Specification

DEFF Research Database (Denmark)

Kalyanova, Olena; Heiselberg, Per

 This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....... This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....

Validation of a serum neutralization test for detection of antibodies specific to cyprinid herpesvirus 3 in infected common and koi carp (Cyprinus carpio).

Science.gov (United States)

Cabon, J; Louboutin, L; Castric, J; Bergmann, S; Bovo, G; Matras, M; Haenen, O; Olesen, N J; Morin, T

2017-05-01

Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of a serious infective, notifiable disease affecting common carp and varieties. In survivors, infection is generally characterized by a subclinical latency phase with restricted viral replication. The CyHV-3 genome is difficult to detect in such carrier fish that represent a potential source of dissemination if viral reactivation occurs. In this study, the analytical and diagnostic performance of an alternative serum neutralization (SN) method based on the detection of CyHV-3-specific antibodies was assessed using 151 serum or plasma samples from healthy and naturally or experimentally CyHV-3-infected carp. French CyHV-3 isolate 07/108b was neutralized efficiently by sera from carp infected with European, American and Taiwanese CyHV-3 isolates, but no neutralization was observed using sera specific to other aquatic herpesviruses. Diagnostic sensitivity, diagnostic specificity and repeatability of 95.9%, 99.0% and 99.3%, respectively, were obtained, as well as a compliance rate of 89.9% in reproducibility testing. Neutralizing antibodies were steadily detected in infected carp subjected to restrictive or permissive temperature variations over more than 25 months post-infection. The results suggest that this non-lethal diagnostic test could be used in the future to improve the epidemiological surveillance and control of CyHV-3 disease. © 2016 John Wiley & Sons Ltd.

Specificities of anti-neutrophil autoantibodies in patients with rheumatoid arthritis (RA)

DEFF Research Database (Denmark)

Brimnes, J; Halberg, P; Jacobsen, Søren

1997-01-01

The objective of this study was to characterize antigens recognized by neutrophil-specific autoantibodies from patients with RA. Sera from 62 RA patients were screened by indirect immunofluorescence (IIF). Positive sera were further tested by ELISAs for antibodies against various granule proteins...

Application of 131I- and 125I-thyroxine in research of thyroid activity control

International Nuclear Information System (INIS)

Michajlovskij, N.; Langer, P.; Stastna, M.; Gschwendtova, L.; Sadlon, J.

1974-01-01

A report is presented on determining the normal levels of free thyroxine and triiodothyronine in the blood sera of man and of animals using tracer techniques with 125 I as a tracer. A method was elaborated of determining thyroxine by equilibrium dialysis and conditions were found of the quantitative chemical analysis of the blood sera of man and of rats for triiodothyronine. A nodification of the method made possible microanalysis of samples smaller than 2 ml, thus permitting the application of the procedure in determining the components in the blood serum of individuals in contrast to the previous practice when only mixtures of blood sera taken from groups of animals could be analyzed. The effect was also studied of certain significant factors, such as anaesthetics and others on the level of free thyroxine in the blood sera of man and of rats. (L.O.)

Inhibition of the mixed leukocyte reaction by alloantisera in man

International Nuclear Information System (INIS)

Jonker, M.; Leeuwen, A. van; Rood, J.J. van

1977-01-01

The incidence of MLC(mixed leukocyte culture)-inhibiting antibodies was determined in 42 pregnancy sera. MLC's were carried out between the cells from the serum donor and her husband in the presence of nonimmune AB serum and the test serum. Fifty per cent of the sera reduced the MLC response to less than 40% of the control values. Only four sera had lymphocytotoxic activity. The inhibition was strong against the specific immunizor, less strong against random unrelated cells and weak against cells which were SD(serologically defined)-identical with the serum donor. Absorptions with lymphocytes and platelets were carried out. Lymphocytes removed activity in three of the four sera tested. Platelets removed activity from one serum. It was concluded that both anti-LD(lymphocyte defined) and anti-SD antibodies were able to inhibit the MLR (mixed leukocyte reaction) at the stimulator cell level. (author)

Humoral immunity through immunoglobulin M protects mice from an experimental actinomycetoma infection by Nocardia brasiliensis.

Science.gov (United States)

Salinas-Carmona, Mario C; Pérez-Rivera, Isabel

2004-10-01

An experimental model of infection with Nocardia brasiliensis, used as an example of a facultative intracellular pathogen, was tested. N. brasiliensis was injected into the rear foot pads of BALB/c mice to establish an infection. Within 30 days, infected animals developed a chronic actinomycetoma infection. Batch cultures of N. brasiliensis were used to purify P61, P38, and P24 antigens; P61 is a catalase, and P38 is a protease with strong caseinolytic activity. Active and passive immunizations of BALB/c mice with these three purified soluble antigens were studied. Protection was demonstrated for actively immunized mice. However, immunity lasted only 30 days. Other groups of immunized mice were bled at different times, and their sera were passively transferred to naive recipients that were then infected with N. brasiliensis. Sera collected 5, 6, and 7 days after donor immunization conferred complete, long-lasting protection. The protective effect of passive immunity decreased when sera were collected 2 weeks after donor immunization. However, neither the early sera (1-, 2-, and 3-day sera) nor the later sera (30- or 45-day sera) prevented the infection. Hyperimmune sera with the highest levels of immunoglobulin G (IgG) to N. brasiliensis antigens did not protect at all. The antigens tested induced two IgM peaks. The first peak was present 3 days after immunization but was not antigen specific and did not transfer protection. The second peak was evident 7 days after immunization, was an IgM response, was antigen specific, and conferred protection. This results clearly demonstrate that IgM antibodies protect the host against a facultative intracellular bacterium.

Incorporation of the ELISA technique to determine antibody levels against foot-and-mouth disease

International Nuclear Information System (INIS)

Vergara, N.N.; Caballero, P.H.; Santiago Gonzalez Patino, S.; Orue, P.M.

1998-01-01

Two groups of sera were evaluated by a liquid phase blocking ELISA (LPBE) for the detection and quantification of foot-and-mouth disease (FMD) antibodies to serotypes O, A and C to assess the sensitivity and specificity of the assay. The first group consisted of 120 sera from non-infected and non-vaccinated cattle, which were tested by a screening assay at a fix dilution of 1/32. The second group consisted of 120 sera from cattle vaccinated with a trivalent (O, A and C) vaccine. Sera from this group were titrated in a five fold dilution range: 1/10, 1/50, 1/250 and 1/1250. (author)

Synthetic Nanovaccines Against Respiratory Pathogens (SYNARP). Addendum

Science.gov (United States)

2014-09-01

laboratory-based influenza surveillance manual (5).To inactivate non-specific inhibitors, sera were treated with receptor destroying enzyme ( RDE ) (Denka...Seiken, Tokyo, Japan) prior to being tested. Briefly, three parts RDE was added to one part sera and incubated overnight at 37°C. RDE was...inactivated by incubation at 56°C for ~60 min. RDE -treated sera were two-fold serially diluted in v-bottom microtiter plates. An equal volume of virus iVN04

IgE antibodies of fish allergic patients cross-react with frog parvalbumin.

Science.gov (United States)

Hilger, C; Thill, L; Grigioni, F; Lehners, C; Falagiani, P; Ferrara, A; Romano, C; Stevens, W; Hentges, F

2004-06-01

The major allergens in fish are parvalbumins. Important immunoglobulin (Ig)E cross-recognition of parvalbumins from different fish species has been shown. Recently frog parvalbumin alpha has been found to be responsible for a case of IgE-mediated anaphylaxis triggered by the ingestion of frog meat. The aim of this study was to investigate whether IgE antibodies of fish allergic persons cross-react with frog parvalbumin and to appreciate its clinical relevance. The sera of 15 fish allergic patients and one fish and frog allergic patient were tested by IgE-immunoblotting against frog muscle extract. Sera were tested against recombinant parvalbumin alpha and beta from Rana esculenta. Skin prick tests were performed in selected patients with recombinant frog parvalbumin. Ca(2+) depletion experiments and inhibition studies with purified cod and frog recombinant parvalbumin were done to characterize the cross-reactive pattern. Fourteen of the sera tested had IgE antibodies recognizing low molecular weight components in frog muscle extract. Calcium depletion experiments or inhibition of patient sera with purified cod parvalbumin led to a significant or complete decrease in IgE binding. When tested against recombinant parvalbumins, three of 13 sera reacted with alpha parvalbumin and 11 of 12 reacted with beta parvalbumin from R. esculenta. Skin prick tests performed with recombinant frog parvalbumin were positive in fish allergic patients. Inhibition studies showed that a fish and frog allergic patient was primarily sensitized to fish parvalbumin. Cod parvalbumin, a major cross-reactive allergen among different fish species, shares IgE binding epitopes with frog parvalbumin. This in vitro cross-reactivity seems to be also clinically relevant. Parvalbumins probably represent a new family of cross-reactive allergens.

Serological studies on the infection of dogs in Ontario with Borrelia burgdorferi, the etiological agent of Lyme disease

Science.gov (United States)

Artsob, Harvey; Barker, Ian K.; Fister, Richard; Sephton, Gregory; Dick, Daryl; Lynch, John A.; Key, Doug

1993-01-01

A serological study was undertaken to determine whether dogs in Ontario are being exposed to Borrelia burgdorferi, the etiological agent of Lyme disease. This study consisted of a survey of randomly selected dogs and testing of diagnostic submissions from candidate Lyme disease cases. The survey of 1,095 dogs, bled between January 1988 and August 1989, revealed a total of 65 (5.9%) enzyme-linked immunosorbent assay (ELISA) reactors, of which 22 had immuno-fluorescent antibody assay (IFA) titers ≥1:32. All but one of the IFA-positive and 10 of the ELISA-positive, IFA-negative sera were further tested by western blot. Eight western blot positive and three equivocal reactors were obtained. Three of the eight confirmed reactors had visited areas known to be endemic for Lyme disease, leaving five reactors that might have been infected in previously undocumented areas for B. burgdorferi activity in Ontario. Diagnostic submissions of sera from 223 dogs were received between August 1987 and February 1992. Test results revealed 21 (9.4%) IFA reactors, of which only six had significant titers (≥1:256) and were reactive by an immunodot Borrelia test. All six dogs had travelled to known Lyme endemic areas. Based on results obtained from this study, it seems likely that the agent of Lyme disease is not widespread in Ontario. PMID:17424284

Previous 60-Co radiation from Paratrygon aiereba mucus induces the production of highly responsive antibodies and a better immune response in mice

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Thomazi, Gabriela Ortega Coelho; Alves, Glaucie Jussilane; Turíbio, Thompson de Oliveira; Rocha, André Moreira; Aires, Raquel da Silva; Jácome, Larissa Barros Silvestre; Spencer, Patrick Jack, E-mail: gabiortegacoelho@usp.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil). Centro de Biotecnologia; Costa, Andrea da; Rodrigues, Jaqueline Pollizeli; Galisteo Júnior, Andrés Jimenez; Andrade Júnior, Heitor Franco de, E-mail: hfandrad@usp.br, E-mail: raquelaires@itpacporto.com.br [Universidade de São Paulo (USP), São Paulo, SP (Brazil). Laboratório de Protozoologia; Seibert, Carla Simone, E-mail: seibertcs@uft.edu.br [Universidade Federal do Tocantins (UFT), Porto Nacional, TO (Brazil)

2017-07-01

Wounds from stinging freshwater stingrays are painful, difficult to heal and cause extensive necrosis and systemic phenomena. The treatment is symptomatic, of low efficiency and there is no therapy, which causes more suffering to the injured. This study aimed to evaluate the immune response induced by the native or irradiated by 60-Co gamma from Paratrygon aiereba mucus. IPEN’s Committee on Ethics in the Use of Animals (n.º126/2013) and lanes captured under license from the Chico Mendes Institute for Biodiversity Conservation (n.º6781-1/2014) approved this research. For the assays, sera from Swiss mice previously immunized against native or irradiated mucus were used. The proliferation of splenic B cells in response to mucus was evaluated by the In Vitro Induced Antibody Production method and serum and splenic cytokines were also quantified. Our data demonstrate that the irradiated mucus of P. aiereba induces greater production of antibodies and more immunological memory in the mice. Spleen cells from animals immunized against irradiated mucus produced IFN-γ, TNF-α and IL-10, and serum TNF-α (immunized group against irradiated mucus) and IL-6 and IL-17 (immunized group against native mucus). The results corroborate the use of ionizing radiation, with production of highly responsive antibodies and better immune response, besides proving that Paratrygon aiereba mucus is capable of stimulating cellular and humoral adaptive immune response, contributing to the continuity of associated investigations. (author)

Previous 60-Co radiation from Paratrygon aiereba mucus induces the production of highly responsive antibodies and a better immune response in mice

International Nuclear Information System (INIS)

Thomazi, Gabriela Ortega Coelho; Alves, Glaucie Jussilane; Turíbio, Thompson de Oliveira; Rocha, André Moreira; Aires, Raquel da Silva; Jácome, Larissa Barros Silvestre; Spencer, Patrick Jack

2017-01-01

Wounds from stinging freshwater stingrays are painful, difficult to heal and cause extensive necrosis and systemic phenomena. The treatment is symptomatic, of low efficiency and there is no therapy, which causes more suffering to the injured. This study aimed to evaluate the immune response induced by the native or irradiated by 60-Co gamma from Paratrygon aiereba mucus. IPEN’s Committee on Ethics in the Use of Animals (n.º126/2013) and lanes captured under license from the Chico Mendes Institute for Biodiversity Conservation (n.º6781-1/2014) approved this research. For the assays, sera from Swiss mice previously immunized against native or irradiated mucus were used. The proliferation of splenic B cells in response to mucus was evaluated by the In Vitro Induced Antibody Production method and serum and splenic cytokines were also quantified. Our data demonstrate that the irradiated mucus of P. aiereba induces greater production of antibodies and more immunological memory in the mice. Spleen cells from animals immunized against irradiated mucus produced IFN-γ, TNF-α and IL-10, and serum TNF-α (immunized group against irradiated mucus) and IL-6 and IL-17 (immunized group against native mucus). The results corroborate the use of ionizing radiation, with production of highly responsive antibodies and better immune response, besides proving that Paratrygon aiereba mucus is capable of stimulating cellular and humoral adaptive immune response, contributing to the continuity of associated investigations. (author)

Neuronal antibodies in patients with suspected or confirmed sporadic Creutzfeldt-Jakob disease.

Science.gov (United States)

Rossi, Meghan; Mead, Simon; Collinge, John; Rudge, Peter; Vincent, Angela

2015-06-01

There have been reports of patients with antibodies to neuronal antigens misdiagnosed as sporadic Creutzfeldt-Jakob disease (sCJD). Conversely, low levels of antibodies to neuronal proteins have been reported in patients with sCJD. However, the frequency of misdiagnoses, or of antibodies in patients with subsequently confirmed sCJD, is not clear. We reviewed 256 consecutive cases of sCJD seen in the National Prion Clinic, of whom 150 had sera previously referred for selected antibody tests. Eighty-two available samples were retested for antibodies to N-methyl-d-aspartate receptor (NMDAR), the glycine receptor (GlyR), voltage-gated potassium channel (VGKC)-complex and the associated proteins, leucine-rich glioma inactivated 1 (LGI1) and contactin-associated protein 2 (CASPR2). Four of the initial 150 sera referred were positive; two had antibodies to NMDAR, and two to the VGKC-complex, one of which was also positive for GlyR antibodies. Of the 82 sCJD sera retested, one had VGKC-complex antibodies confirming the previous result, two had CASPR2 and GlyR antibodies and one had CASPR2 and NMDAR antibodies; all antibodies were at low levels. Over the same period three patients with autoimmune encephalitis and high VGKC-complex antibodies were initially referred as sCJD. This study indicates that VGKC-complex/LGI1 antibodies. Low titres of neuronal antibodies occur only rarely in suspected patients with sCJD and when present should be interpreted with caution. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Phospholipids in sera of horses with summer eczema: lipid analysis of the autoserum preparation used in therapy.

Science.gov (United States)

Hallamaa, R E; Batchu, K C; Tallberg, T

2014-05-01

Equine summer eczema, also known as insect bite hypersensitivity, affects horses recurrently during summer months. The treatment of this allergic pruritus is difficult and therefore there is a need for efficacious treatments. Autoserum therapy, based on the use of autogenous serum that is specifically prepared for oral administration and given when the animal shows clinical signs has been introduced recently. Lipids are thought to be responsible for the effect of this therapy. The main aim of this study was to analyse the phospholipid content of autogenous serum preparations and to further assess whether these preparations have different lipid profiles depending on the clinical status of the horse. The hypothesis is that the major serum phospholipids typical of the horse are present in the autoserum preparation. Descriptive controlled clinical study. Sera were collected from 10 affected and 6 healthy horses, prepared in a similar fashion and the lipids contained in the resulting autoserum preparations were analysed by electrospray ionisation mass spectrometry. The major phospholipid classes detected were phosphatidylcholine, sphingomyelin, phosphatidic acid and traces of lysophosphatidylcholine. Horses with summer eczema had significantly abundant concentrations of phosphatidylcholine (P = 0.042) and sphingomyelin (P = 0.0017) in comparison with healthy horses, while the concentration of phosphatidic acid was significantly higher in healthy horses (P = 0.0075). The autoserum preparation contains minute amounts of the main serum phospholipids in differing concentrations in healthy horses and horses with an allergic skin disease. © 2013 EVJ Ltd.

Assessment of reactivity of three treponemal tests in non-treponemal non-reactive cases from sexually transmitted diseases clinic, antenatal clinic, integrated counselling and testing centre, other different outdoor patient departments/indoor patients of a tertiary care centre and peripheral health clinic attendees

Directory of Open Access Journals (Sweden)

M Bala

2013-01-01

Full Text Available In India, many state reference centres for sexually transmitted infections perform only a single screening assay for syphilis diagnosis. In this study, Treponema pallidum haemagglutination (TPHA was performed on 1115 Venereal Disease Research Laboratory (VDRL/rapid plasma regain (RPR non-reactive and 107 reactive sera out of 10,489 tested by VDRL/RPR according to the National AIDS Control Organisation syphilis testing protocol. A total of 47 Specimens reactive in TPHA and non-reactive with VDRL test were subjected to fluorescent treponemal antibody absorption and enzyme-immunoassay. Seroprevalence considering both VDRL and TPHA positivity was highest (4.4% in sexually transmitted diseases clinic attendees than in other subject groups. Positivity by two treponemal tests in 24 (2.2% cases non-reactive by VDRL/RPR was representative of the fully treated patients or latent or late syphilis cases. The findings highlight that a suitable treponemal confirmatory test should be performed in all the diagnostic laboratories.

Interference from previous distraction disrupts older adults' memory.

Science.gov (United States)

Biss, Renée K; Campbell, Karen L; Hasher, Lynn

2013-07-01

Previously relevant information can disrupt the ability of older adults to remember new information. Here, the researchers examined whether prior irrelevant information, or distraction, can also interfere with older adults' memory for new information. Younger and older adults first completed a 1-back task on pictures that were superimposed with distracting words. After a delay, participants learned picture-word paired associates and memory was tested using picture-cued recall. In 1 condition (high interference), some pairs included pictures from the 1-back task now paired with new words. In a low-interference condition, the transfer list used all new items. Older adults had substantially lower cued-recall performance in the high- compared with the low-interference condition. In contrast, younger adults' performance did not vary across conditions. These findings suggest that even never-relevant information from the past can disrupt older adults' memory for new associations.

Detection of anti neutrophil antibodies by radio-iodinated protein A binding test

International Nuclear Information System (INIS)

Cartron, J.; Muller, J.Y.; Tchernia, G.; Paule, B.; Varet, B.

1983-01-01

The granulocyte associated IgG in normal and neutropenic subjects has been determined by a direct quantitative assay using radiolabeled staphylococcal protein A. This assay allows to postulate an immunological mechanism to explain the neutropenia in 19 cases of neutropenias associated with malfunctions of the immune system and in 4 cases of idiopathic neutropenias. Discussed in this report is the possible interaction of immune complexes bound in vivo to the granulocytes. By an immunofluorescence test, it has been possible to detect IgG or IgM antibodies in only 52% of patients with a positive direct assay. The determination of granulocyte-associated IgG is therefore a better indicator for defining an auto-immune neutropenia than the detection of free antibodies in the sera [fr

Differential recognition of terminal extracellular Plasmodium falciparum VAR2CSA domains by sera from multigravid, malaria-exposed Malian women.

Science.gov (United States)

Travassos, Mark A; Coulibaly, Drissa; Bailey, Jason A; Niangaly, Amadou; Adams, Matthew; Nyunt, Myaing M; Ouattara, Amed; Lyke, Kirsten E; Laurens, Matthew B; Pablo, Jozelyn; Jasinskas, Algis; Nakajima, Rie; Berry, Andrea A; Takala-Harrison, Shannon; Kone, Abdoulaye K; Kouriba, Bourema; Rowe, J Alexandra; Doumbo, Ogobara K; Thera, Mahamadou A; Laufer, Miriam K; Felgner, Philip L; Plowe, Christopher V

2015-06-01

The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family mediates parasite sequestration in small capillaries through tissue-specific cytoadherence. The best characterized of these proteins is VAR2CSA, which is expressed on the surface of infected erythrocytes that bind to chondroitin sulfate in the placental matrix. Antibodies to VAR2CSA prevent placental cytoadherence and protect against placental malaria. The size and complexity of the VAR2CSA protein pose challenges for vaccine development, but smaller constitutive domains may be suitable for subunit vaccine development. A protein microarray was printed to include five overlapping fragments of the 3D7 VAR2CSA extracellular region. Malian women with a history of at least one pregnancy had antibody recognition of four of these fragments and had stronger reactivity against the two distal fragments than did nulliparous women, children, and men from Mali, suggesting that the C-terminal extracellular VAR2CSA domains are a potential focus of protective immunity. With carefully chosen sera from longitudinal studies of pregnant women, this approach has the potential to identify seroreactive VAR2CSA domains associated with protective immunity against pregnancy-associated malaria. © The American Society of Tropical Medicine and Hygiene.

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

Seroprevalence of tick-borne-encephalitis virus in wild game in Mecklenburg-Western Pomerania (north-eastern Germany).

Science.gov (United States)

Frimmel, Silvius; Leister, Matthias; Löbermann, Micha; Feldhusen, Frerk; Seelmann, Matthias; Süss, Jochen; Reisinger, Emil Christian

2016-10-01

Mecklenburg-Western Pomerania, a federal state in the north east of Germany, has never been a risk area for TBEV infection, but a few autochthonous cases, along with TBEV-RNA detection in ticks, have shown a low level of activity in natural foci of the virus in the past. As wild game and domestic animals have been shown to be useful sentinels for TBEV we examined sera from wild game shot in Mecklenburg-Western Pomerania for the prevalence of TBEV antibodies. A total of 359 sera from wild game were investigated. All animals were shot in Mecklenburg-Western Pomerania in 2012. Thirteen of 359 sera tested positive or borderline for anti-TBEV-IgG with ELISA and four samples tested positive using NT. The four TBEV-positive sera confirmed by NT constitute the first detection of TBEV-antibodies in sera of wild game in Mecklenburg-Western Pomerania since 1986-1989. This underlines that the serological examination of wild game can be a useful tool in defining areas of possible TBEV infection, especially in areas of low TBEV-endemicity. Copyright © 2016 Elsevier GmbH. All rights reserved.

Measurement of anti-IgA antibodies by a two-site immunoradiometric assay

International Nuclear Information System (INIS)

Homburger, H.A.; Smith, J.R.; Jacob, G.L.; Laschinger, C.; Naylor, D.H.; Pineda, A.A.

1981-01-01

To enable the detection of IgG class, anti-IgA antibodies and to investigate the possible occurrence of IgE class, anti-IgA antibodies, we developed a solid phase immunoradiometric assay, which uses purified IgA coupled covalently to microcrystalline cellulose as an immunosorbent. Radiolabeled, Fc specific anti-IgG and anti-IgE antibodies were used to detect specific aIgA after incubation of test sera or controls with the immunosorbent. IgG-aIgA were detected by the IRA in 100 and 67% of control sera with class specific and limited specificity aIgA. The IRA was sensitive to approximately two ng of class specific IgG-aIgA. IgG-aIgA also were detected by IRA in 7.9% of sera from patients with urticarial transfusion reactions and 73% of sera from patients with ataxia telangiectasia and IgA deficiency. Sera from 50 normal blood donors did not have detectable IgG-aIgA. Tests for IgE-aIgA were negative in all cases, including control sera with class specific IgG-aIgA. We conclude that the IRA is a sensitive and reproducible method for detection of class specific and limited specificity IgG-aIgA, and that IgE-aIgA do not mediate urticarial transfusion reactions

Radioimmunoassay in the detection of the hepatitis Be antigen/antibody system in asymptomatic carriers of hepatitis B surface antigen

International Nuclear Information System (INIS)

Pastore, G.; Dentico, P.; Angarano, G.; Schiraldi, O.; Zanetti, A.R.; Ferroni, P.

1980-01-01

A radioimmunoassay for hepatitis e antigen (HBeAg) and antibody to e (anti-HBe) was developed and sera of 71 asymptomatic chronic carriers of hepatitis B surface antigen (HBsAg), in 44 of whom liver biopsy was obtained, were tested. In addition, testing for Dane particle associated DNA polymerase activity was performed in all sera. HBeAg was detected in 14 subjects (19.7%) and anti-HBe in 46 (64.8%). The highest proportion of HBeAg positivity (40%) was found among carriers with histological evidence of chronic hepatitis, whereas anti-HBe was present in 80% of carriers with normal liver histology, in 58% of carriers with non-specific reactive hepatitis and in 60% of carriers with chronic liver lesions. DNA polymerase activity was present in 92.8% of sera positive for HBeAg, in 13% of sera positive for anti HBe, and in 9% of sera negative for both markers. Our results demonstrate that not all HBsAg carriers reactive to HBeAg show evidence of chronic hepatitis nor, conversely, that anti-HBe is invariably associated with the healthy carrier state of HBsAg. Finally, circulating Dane particles, as revealed by the presence of serum specific DNA polymerase activity, may also be present in anti-HBe positive sera other than those of some HBsAg carriers lacking both HBeAg and anti-HBe. (orig.) [de

Assessing drivers of the IgG4 antibody reactivity to recombinant antigen Bm14 in Wuchereria bancrofti endemic populations in East Africa

DEFF Research Database (Denmark)

Hansen, Johanne Damgaard; Meyrowitsch, Dan W.; Rwegoshora, Rwehumbiza T.

2016-01-01

to the recombinant filarial antigen Bm14 in Wuchereria bancrofti endemic populations in East Africa. Sera collected during previous studies from 395 well characterized individuals with regard to age, sex, mf, CFA, household vector biting and household exposure to infective filarial larvae were tested for IgG4......A high proportion of the human population in lymphatic filariasis (LF) endemic areas is positive for filarial specific IgG4 antibodies, including many individuals without microfilariae (mf; circulating larvae in the human blood) or circulating filarial antigens (CFA; marker of adult worm infection...

Evaluation of recombinant porin (rOmp2a) protein as a potential antigen candidate for serodiagnosis of Human Brucellosis.

Science.gov (United States)

Pathak, Prachi; Kumar, Ashu; Thavaselvam, Duraipandian

2017-07-11

Brucellosis is an important zoonotic disease caused by different Brucella species and human brucellosis is commonly prevalent in different states of India. Among various Brucella species, B. melitensis is most pathogenic to human and included as category B biothreat which can cause infection through aerosol, cut, wounds in skin and contact with infected animals. The diagnosis of human brucellosis is very important for proper treatment and management of disease as there is no vaccine available for human use. The present study was designed to clone, express and purify immunodominant recombinant omp2a (rOmp2a) porin protein of B. melitensis and to evaluate this new antigen candidate for specific serodiagnosis of human brucellosis by highly sensitive iELISA (indirect enzyme linked immunosorbent assay). Omp2a gene of B. melitensis 16 M strain was cloned and expressed in pET-SUMO expression system. The recombinant protein was purified under denaturing conditions using 8 M urea. The purified recombinant protein was confirmed by western blotting by reacting with anti-HIS antibody. The sero-reactivity of the recombinant protein was also checked by reacting with antisera of experimentally infected mice with B. melitensis 16 M at different time points. Serodiagnostic potential of recombinant porin antigen was tested against 185 clinical serum samples collected from regions endemic to brucellosis in southern part of India by iELISA. The samples were grouped into five groups. Group 1 contained cultured confirmed positive serum samples of brucellosis (n = 15), group 2 contained sera samples from positive cases of brucellosis previously tested by conventional methods of RBPT (n = 28) and STAT (n = 26), group 3 contained sera samples negative by RBPT(n = 36) and STAT (n = 32), group 4 contained sera samples of other febrile illness and PUO case (n = 35) and group 5 contained confirmed negative sera samples from healthy donors (n = 23). The rOmp2a was found to be

Borrelia burgdorferi antibodies in dogs from Cotia county, São Paulo State, Brazil

Directory of Open Access Journals (Sweden)

JOPPERT Adriana Marques

2001-01-01

Full Text Available Dogs sera samples collected from Cotia County, São Paulo were tested using indirect immunoenzymatic test (ELISA in order to study Lyme disease serology in dogs. ELISA method was standardized and G39/40 North American strain of Borrelia burgdorferi was used as antigen. Positive results were confirmed employing the Western blotting technique. Because of the possibility of cross-reactions, sera were also tested for different serological strains of Leptospira interrogans and L. biflexa using microscopic sera agglutination test. Twenty-three of 237 (9.7% serum samples were positive in the ELISA; 20 of them (86.9% were confirmed by the Western blotting, what suggests that Cotia may be a risk area for Lyme disease. Although 4 samples (1.7% were positive for Lyme disease and leptospirosis, no correlation was found between the results (X² = 0.725; p = 0.394 what suggests absence of serological cross reactivity.

Cross reactivities of rabbit anti-chicken horse radish peroxidase ...

African Journals Online (AJOL)

The cross reactivities of rabbit anti chicken horse radish peroxidase (conjugate) was tested with sera of Chicken, Ducks, Geese, Guinea fowl, Hawks, Pigeons and Turkeys in indirect enzyme linked immunosorbent assay (ELISA) technique. Sera from mammalian species (Bat, Equine and swine) were used as negative ...

Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus

DEFF Research Database (Denmark)

Brocchi, E.; Bergmann, I.E.; Dekker, A.

2006-01-01

To validate the use of serology in substantiating freedom from infection after foot-and-mouth disease (FMD) outbreaks have been controlled by measures that include vaccination, 3551 sera were tested with six assays that detect antibodies to the non-structural proteins of FMD virus. The sera came...

Immune Infertility Should Be Positively Diagnosed Using an Accurate Method by Monitoring the Level of Anti-ACTL7a Antibody

Science.gov (United States)

Fu, Jun; Yao, Rongyan; Luo, Yanyun; Yang, Dantong; Cao, Yang; Qiu, Yi; Song, Wei; Miao, Shiying; Gu, Yiqun; Wang, Linfang

2016-01-01

Infertility is currently a major public health problem. Anti-sperm antibodies (ASAs) markedly reduce sperm quality, which can subsequently lead to male and/or female infertility. The accurate detection of ASAs derived from specific spermatozoa is, therefore, clinically useful. We have focused on the spermatozoa-specific expression protein ACTL7a for many years and have developed an enzyme-linked immunosorbent assay (ELISA) to detect the concentration of anti-ACTL7a antibodies in fertile sera (n = 267) and infertile sera (n = 193). Infertile sera were collected from the positive sera of tray agglutination tests (TAT), which is a routine ASA screening methodology. We found that the concentration of anti-ACTL7a antibodies was significantly higher in the infertile sera (than in the fertile sera, P infertile sera. The ELISA was much better for male sera detection (AUC = 0.9899). If we set the standard at a strongly positive value (calculated by ROC curve), the positive predictive value of the antibody detection reached 100 percent, with a false positive rate of zero. The developed ELISA method for anti-ACTL7a antibody detection is therefore sensitive, accurate, and easy to perform, making it an excellent potential tool for future clinical use. PMID:26957350

Evaluation of the Leptospira species microscopic agglutination test in experimentally vaccinated cats and Leptospira species seropositivity in aged azotemic client-owned cats.

Science.gov (United States)

Shropshire, Sarah B; Veir, Julia K; Morris, Arianne K; Lappin, Michael R

2016-10-01

The objectives of this study were to validate the microscopic agglutination test (MAT) using feline sera, determine cross-reactivity of Borrelia burgdorferi antibodies in the MAT, and evaluate if there is an association between Leptospira species seropositivity in aged (⩾10 years) client-owned cats with and without azotemia (creatinine >2 g/dl). A four-serovar canine leptospiral vaccine was administered to two specific pathogen-free (SPF) cats on days 0 and 14. The MAT was performed intermittently until day 42 for the serovars Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Bratislava, with a cut-off value of ⩾1:100. Five purpose-bred cats were infested with wild-caught Ixodes scapularis adults with an average B burgdorferi infection rate of 50%, and tested for antibodies against B burgdorferi C6 peptide and DNA in skin biopsies, as well as by MAT. Sera from 66 azotemic and 75 non-azotemic cats ⩾10 years of age were tested for Leptospira species antibodies using the MAT and results were compared by the χ(2) test. Both SPF cats seroconverted by week 3 and formed antibodies against at least one serovar. There was no cross-reactivity in the MAT using samples from cats with antibodies to B burgdorferi. MAT results were positive for 4/66 azotemic cats and 8/75 non-azotemic cats; these results were not statistically different. The MAT can be interpreted using feline serum and does not appear to cross-react in cats with B burgdorferi antibodies. There was no association between Leptospira species MAT results and azotemia in this group of aged client-owned cats but further studies are needed to determine if leptospirosis contributes to feline chronic kidney disease. © The Author(s) 2015.

Gaze Stabilization Test Asymmetry Score as an Indicator of Previous Concussion in a Cohort of Collegiate Football Players.

Science.gov (United States)

Honaker, Julie A; Criter, Robin E; Patterson, Jessie N; Jones, Sherri M

2015-07-01

Vestibular dysfunction may lead to decreased visual acuity with head movements, which may impede athletic performance and result in injury. The purpose of this study was to test the hypothesis that athletes with history of concussion would have differences in gaze stabilization test (GST) as compared with those without a history of concussion. Cross-sectional, descriptive. University Athletic Medicine Facility. Fifteen collegiate football players with a history of concussion, 25 collegiate football players without a history of concussion. Participants completed the dizziness handicap inventory (DHI), static visual acuity, perception time test, active yaw plane GST, stability evaluation test (SET), and a bedside oculomotor examination. Independent samples t test was used to compare GST, SET, and DHI scores per group, with Bonferroni-adjusted alpha at P history of concussion. The results support further research on the use of GST for sport-related concussion evaluation and monitoring. Inclusion of objective vestibular tests in the concussion protocol may reveal the presence of peripheral vestibular or visual-vestibular deficits. Therefore, the GST may add an important perspective on the effects of concussion.