Sample records for antinuclear antibody screen

  1. [Antinuclear antibodies]. (United States)

    Cabiedes, Javier; Núñez-Álvarez, Carlos A


    Anti-nuclear antibodies (ANA) are immunoglobulin directed against autologous cell nuclear and cytoplasmic components. Besides the autoimmune ANA there are other ANA that can be detected in circulation, like natural and infectious ANA. Because of its high sensibility, detection of the ANA must be done by indirect immunofluorescence (IIF) as screening test and all of those positive samples are convenient to confirm its specificity by ELISA, western blot or other techniques. Positive ANA detected by IIF must be evaluated taking in to account the pattern and titer. The following recommended step is the specificity characterization (reactivity against extractable nuclear antigens [ENA], dsDNA, etc.) which is useful for the diagnosis and follow up of patients with autoimmune diseases, and by such reasoning, its detection must be performed in an orderly and reasonable way using guides or strategies focused to the good use and interpretation of the autoantibodies. The objective of this review is to present a compilation of the literature and our experience in the detection and study of the ANA.

  2. Antinuclear antibody panel (United States)

    ... page: // Antinuclear antibody panel To use the sharing features on this page, please enable JavaScript. The antinuclear antibody panel is a blood test that looks at ...

  3. Evaluation of the LIAISON ANA screen assay for antinuclear antibody testing in autoimmune diseases. (United States)

    Ghillani, P; Rouquette, A M; Desgruelles, C; Hauguel, N; Le Pendeven, C; Piette, J C; Musset, L


    Antinuclear antibodies (ANA) are widely detected by immunofluorescence on HEp-2 cells in patients with connective tissue diseases and other pathological conditions. We evaluated the first-automated chemiluminescence immunoassay for the detection of ANA (LIAISON ANA screen, DiaSorin). This study was carried out simultaneously in two laboratories by testing 327 patient samples with clinically defined connective diseases, 273 routine samples for ANA screening, and 300 blood donors. A total of 268 out of 337 IIF-positive sera were positive with LIAISON ANA screen (79.5% of agreement) and 240 out of 263 IIF-negative sera were negative with LIAISON ANA screen (91.2% of agreement). After resolution of discrepant results, the concordance reached, respectively, 94.9% and 98.8%. The specificity was 99.3% and the sensitivity was 94%. Unlike results obtained by other ANA screening assays, we observed acceptable sensitivity and specificity. Despite the presence of HEp-2 cell extract, we failed to detect some antibodies as antinucleolar, antinuclear envelope, and antiproliferating cell nuclear antigen. This automated assay allows quick process to results and exhibits satisfactory sensitivity for the detection of the main ANA specificities of connective tissue diseases.

  4. Evaluation of the BioPlex 2200 ANA screen for the detection of antinuclear antibodies and comparison with conventional methods. (United States)

    Desplat-Jego, Sophie; Bardin, Nathalie; Larida, Bruno; Sanmarco, Marielle


    BioPlex 2200 multiplexed assays system is an automatic method allowing detection of antinuclear antibodies (ANA). The aim of our study was to evaluate the determination of 13 autoantibodies against chromatinic and nonchromatinic nuclear antigens by the BioPlex 2200 system and to compare the results achieved by this method to those obtained with our routinely used immunoassays. One thousand and four serum samples consecutively sent for ANA detection were routinely tested by indirect immunofluorescence (IIF) on HEp2 cells. Among them, 321 were also analyzed by dsDNA enzyme immunoassay (EliA) test and 657 by double immunodiffusion (DID) for extractable nuclear antigen (ENA) antibodies. All the sera were evaluated by the BioPlex 2200 ANA screen kit allowing simultaneous detection of antibodies against the following antigens: dsDNA, chromatin, SSA-52 kDa, SSA-60 kDa, SSB, Sm, Sm/RNP, RNP-A, RNP-68 kDa, Scl70, centromere B, Jo-1, and P ribosomal proteins. The kappa coefficient between BioPlex 2200 and routine tests for detection of ANA on HEp2 cells, anti-dsDNA, and anti-ENA antibodies was, respectively, 0.31, 0.66, and 0.61. The comparison with our routine tests showed numerous discrepancies between IIF ANA screening and BioPlex but a good concordance for detection of anti-dsDNA and anti-ENA specificities. BioPlex 2200 system is a rapid and sensitive method for simultaneous quantitative detection of several autoantibodies. It is perfectly well adapted to determine ANA antigenic specificities of samples found positive using initial IIF screening. The capability of this multiplexed technology to analyze simultaneously 13 ANA autoantibodies leads to the rapid availability of an "autoimmune connective tissue disease serologic profile."

  5. Systemic sclerosis without antinuclear antibodies or Raynaud's phenomenon

    DEFF Research Database (Denmark)

    Schneeberger, D.; Tyndall, A.; Walker, U.A.;


    Objective: To assess patients with SSc who present without circulating antinuclear antibodies (ANA) or Raynaud’s phenomenon (RP). Methods: 5390 patients who fulfilled the ACR criteria for SSc and were enrolled in the EULAR Scleroderma Trials And Research (EUSTAR) database were screened...

  6. Antinuclear antibodies in scleroderma, mixed connective tissue disease and "primary" Raynaud's phenomenon. (United States)

    Cruz, M; Mejia, G; Lavalle, C; Cortes, J J; Reyes, P A


    The diversity of antibodies in patients with scleroderma, mixed connective tissue disease or "primary" Raynaud's phenomenon could be used as a laboratory aid in the clinical diagnosis. In serum samples of 75 patients we screened for antinuclear antibodies (HEp 2 cells), anti DNA, soluble nucleoprotein and extractable nuclear antigens (Sm, rRNP, U1-nRNP, SSA/Ro, SSB/La and Scl-70). Distinctive antinuclear antibodies pattern was identified in each group of patients. This immunologic profile is valuable for clinical diagnosis and the preferential association of certain autoantibodies with some diseases and not with others, suggest an antigen-driven stimulus for its production.

  7. Diagnostic accuracy of ELISA methods as an alternative screening test to indirect immunofluorescence for the detection of antinuclear antibodies. Evaluation of five commercial kits. (United States)

    Tonuttia, Elio; Bassetti, Danila; Piazza, Anna; Visentini, Daniela; Poletto, Monica; Bassetto, Franca; Caciagli, Patrizio; Villalta, Danilo; Tozzoli, Renato; Bizzaro, Nicola


    Detection of antinuclear antibodies (ANA) is a fundamental laboratory test for diagnosing systemic autoimmune diseases. Currently, the method of choice is indirect immunofluorescence (IIF) on a HEp-2 cell substrate. The goal of this study was to evaluate the diagnostic accuracy of five commercially available enzyme immunoassay (EIA) kits for ANA detection and to verify the possibility of using them as an alternative to the IIF method. The study involved 1513 patients, 315 of whom were diagnosed with a systemic autoimmune disease and 1198 in whom an autoimmune disorder was excluded. For all sera, ANA detection was performed via IIF and with five different EIA kits. The results were evaluated in relation to clinical diagnosis and the presence of possible specific autoantibodies (anti-ENA or anti-dsDNA); lastly, they were compared with the results obtained using ANA-IIF as the method of reference. The positive rate of the ANA-IIF test in subjects with systemic autoimmune diseases was 92%, whereas in the five ANA-EIA kits there was broad diversity in terms of response, with positive rates ranging from 74 to 94%. All the EIA kits correctly detected the presence of antibodies (anti-dsDNA, anti-RNP, anti-Ro/SSA) responsible for homogeneous and speckled fluorescence pattern, but at the same time they showed substantial inaccuracy with the nucleolar pattern, with a mean sensitivity of approximately 50% in this case. Instead, there was a large kit-to-kit difference in terms of identification of anti-Scl70 and centromere patterns, for which sensitivities ranged between 45 and 91%, and between 49 and 100%, respectively. The results of the study demonstrate that the commercially available ANA-EIA kits show different levels of sensitivity and specificity. Some of them have a diagnostic accuracy that is comparable and, in some cases, even higher than the IIF method. Consequently, these could be used as an alternative screening test to IIE. However, others do not ensure acceptable

  8. 21 CFR 866.5100 - Antinuclear antibody immunological test system. (United States)


    ... autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antinuclear antibody immunological test system. 866.5100 Section 866.5100 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  9. 抗核抗体筛查试验与特异性抗体确认试验的相关性研究%Correlation between screening assay of antinuclear antibody and confirmatory assay of specific antibody

    Institute of Scientific and Technical Information of China (English)

    周仁芳; 胡朝军; 张蜀澜; 白伊娜; 宋宁; 李丽君; 李永哲


    Objective To investigate the correlation between screening assay of antinuclear antibody ( ANA ) and confirmatory assay of specific antibody. Methods Five hundred clinical patients' serum samples were detected by indirect immunofluorescence ( I1F) for screening assay of ANAs and line immunoassay ( LIA) for confirmatory assay of specific ANAs. According to the results of IIF-ANA and LIA-ANAs, 500 samples were divided into 4 groups; IIF-ANA ~+/LIA-ANAs~+ group, IIF-ANA~+/LIA-ANAs~-group, IIF-ANA~-/LIA-ANAs~+ group and IIF-ANA~-LIA-ANAs~- group. The samples in IIF-ANA~-VLIA-ANAs~+ group were collected and rechecked by ELISA and DID for detection of specific ANAs. Results Five hundred clinical samples were identified including 247 cases of IIF-ANA~+/LJA-ANAs~+86 cases of IIF-ANA ~+/LIA-ANAs~- , 152 cases of IIF-ANA~-/LIA-ANAs~- and 15 cases of IIF-ANA~- /LIA-ANAs~+, In IIF-ANA ~+/LIA-ANAs~+ group, 48. 2% ,23. 9% and 27. 9% patients had the titre of IIF-ANA 1:1 280, 1: 80-1:160 and 1:320-1:640. Fifteen cases of LJA-ANAs~+ were identified in 167 cases of HF-ANA(9. 0% ). Among 15 cases of IIF-ANA~-/LIA-ANAs~+ 11 positive cases were identified as anti-SSA and anti-Ro52 positive, which were rechecked by ELISA and DID, and the results were consistent with the prior detection. Other positive cases were identified including 1 case of anti-Histone, 1 case of anti-Scl-70, 1 case of anti-Jo-1 and 1 case of anti-AMA-M2. The positive consistent rates between LJA-ANAs and ELJSA, DID were 100% , 69% respectively. And 8 cases of IIF-ANA~-/LJA-ANAs~+ patients were diagnosed with autoimmune disease after clinical data analysis. Among 86 cases of IIF-ANA ~+ /LJA-ANAs~- , 50.0% and 36.1% patients had the titre of 1: 80-1: 160 and homogeneous fluorescence pattern respectively. There was a significant difference in IIF-ANA~+/LIA-ANAs~- group and IIF-ANA ~+/LIA-ANAs~+ (χ~2 =20.47,12.42, P<0. 05). Conclusions It is recommend that specific antinuclear antibodies can be used once screening

  10. Antinuclear antibody-negative systemic lupus erythematosus-how common?



    A review of five years' DNA-binding antibody results in a routine service laboratory revealed 38 patients who had high DNA-binding capacity (DNA-bc) but no antinuclear antibodies (ANA). On retrospective case note analysis, 22 patients (58%) were thought to have systemic lupus erythematosus (SLE), although only six (16%) fulfilled the preliminary classification criteria of the American Rheumatism Association (ARA). Our findings indicate that ANA-negative SLE is commoner than generally realised...

  11. Antinuclear Antibody-Positive Ticlopidine-Induced Hepatitis

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    Sander Jo Veldhuyzen van Zanten


    Full Text Available Ticlopidine hydrochloride has been shown to reduce the risk of first or recurrent stroke in patients who have experienced a transient ischemic attack, reversible ischemic neurological deficit, recurrent stroke or first stroke. Severe liver dysfunction is a contraindication for its use. Increase in liver enzymes has been reported with use of this drug, but jaundice is rare. A case of severe ticlopidine-induced hepatitis that was associated with a marked increase in antinuclear antibody (ANA levels is reported. Physicians prescribing ticlopidine hydrochloride should be aware that a potentially severe acute hepatitis associated with ANA positivity can occur. The drug should be discontinued if signs of liver dysfunction occur.

  12. Clinical utility of antinuclear antibody tests in children

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    Kickingbird Lauren M


    Full Text Available Abstract Background Antinuclear antibody (ANA tests are frequently used to screen children for chronic inflammatory diseases such as systemic lupus erythematosus (SLE. However, the diagnostic utility of this test is limited because of the large number of healthy children who have low-titer positive tests. We sought to determine the clinical utility of ANA tests in screening children for rheumatic disease and to determine whether there are specific signs or symptoms that enhance the clinical utility of ANA tests in children. Methods We undertook a retrospective analysis of 509 new patient referrals. Charts of patients referred because of results of ANA testing were selected for further analysis. Children with JRA, SLE, and other conditions were compared using demographic data, chief complaints at the time of presentation, and ANA titers. Results One hundred ten patients were referred because of an ANA test interpreted as positive. Ten patients were subsequently diagnosed with SLE. In addition, we identified one patient with mixed connective tissue disease, and an additional child with idiopathic Raynaud's phenomenon. Eighteen children of the children referred for a positive ANA test had juvenile rheumatoid arthritis (JRA. Another 80 children with positive ANA tests were identified, the majority of whom (n = 39, 49% had musculoskeletal pain syndromes. Neither the presence nor the titer of ANA served to distinguish children with JRA from children with other musculoskeletal conditions. Children with JRA were readily identified on the basis of the history and physical examination. Children with SLE were therefore compared with children with positive ANA tests who did not have JRA, designated the "comparison group." Non-urticarial rash was more common in children with SLE than in children without chronic inflammatory disease (p = 0.007. Children with SLE were also older (mean ± sd = 14.2 ± 2.5 years than the comparison group (11.0 ± 3.6 years; p

  13. High Prevalence of Antinuclear Antibodies in Children with Thyroid Autoimmunity

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    Maria Segni


    Full Text Available Background. Antinuclear antibodies (ANA are a hallmark of many autoimmune diseases and can be detected many years before disease onset. Autoimmune thyroid diseases (AITD are frequently associated with other organ- and non-organ-specific autoimmune disorders. Objectives. To assess the prevalence of ANA in pediatric patients with AITD and their clinical correlations. Methods. Ninety-three consecutive pediatric patients with AITD were enrolled (86 children with chronic lymphocytic thyroiditis and 7 with Graves’ disease. ANA, anti-double DNA (anti-dsDNA antibodies, anti-extractable nuclear antigen (anti-ENA, anti-cyclic citrullinated peptide antibodies (anti-CCP, and rheumatoid factor (RF was obtained. Signs and symptoms potentially related to rheumatic diseases in children were investigated by a questionnaire. Results. ANA positivity was found in 66/93 children (71%, anti-ENA in 4/93 (4.3%, anti-dsDNA in 1/93 (1.1%, RF in 3/93 (3.2%, and anti-CCP in none. No significant differences were found between the ANA-positive and ANA-negative groups with respect to age, sex, L-thyroxine treatment, or prevalence of other autoimmune diseases. Overall, parental autoimmunity was found in 23%. Conclusions. ANA positivity was demonstrated in 71% of children with AITD. ANA positivity was not related to overt immune-rheumatic diseases. However, because the positivity of ANA can occur even many years before the onset of systemic autoimmune diseases, prospective studies are warranted.

  14. Autoimmune thyroiditis in antinuclear antibody positive children without rheumatologic disease

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    Arkachaisri Thaschawee


    Full Text Available Abstract Background Children are commonly referred to a pediatric rheumatology center for the laboratory finding of an Anti-nuclear antibody (ANA of undetermined significance. Previous studies regarding adult rheumatology patients have supported an association between ANA and anti-thyroid antibodies, with the prevalence of thyroid antibodies being significantly higher in patients referred to a rheumatology center for an ANA without evidence of connective tissue disease compared to the general population. The purpose of the present study was to determine the frequency of thyroid antibodies in children referred to a pediatric rheumatology center for a positive ANA without evidence of a connective tissue disease. Methods A retrospective chart review was performed on children who were referred to our pediatric rheumatology center between August 2003 and March 2007 for positive ANA with concurrent thyroid antibody and thyroid function tests performed who did not fulfill criteria for a specific connective tissue disease. Laboratory and clinical features were recorded and analyzed. Mean and standard deviation were used to describe continuous data. Chi-square or Fisher's exact tests were used to compare proportions between variables. Results One-hundred and four ANA-positive patients with concurrent thyroid studies were evaluated (88% female, 93% Caucasian, mean age 11.9 ± 4.0 years. Half of patients had an ANA titer ≥ 1:320. The ANA pattern was speckled in 60% of the patients. Thyroid antibodies were detected in 30% of the patients. Anti-Thyroglobulin (ATG was detected in 29% and Anti-thyroid peroxidase (ATPO in 21% of the patients; of these children, 14% had hypothyroidism. ANA pattern and titer were not associated with anti-thyroid antibody positivity. Conclusion Thyroid antibodies associated with chronic lymphocytic thyroiditis, ATG and ATPO, were detected significantly higher in ANA-positive children without a rheumatologic condition (30% as

  15. A case of metabolic syndrome, positive antinuclear antibodies and cirrhosis.

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    Alessandro Grasso


    Full Text Available Metabolic syndrome is a common clinical condition often associated with hepatic steatosis and sometimes with mild transaminases and gamma-glutamyl transpeptidase increase. We report a case of a 45-years-old female, with mild increase of ALT, who was followed-up in our Department. A liver biopsy was then performed when an atorvastatin-induced ALT flare, as well as a positive antinuclear antibody appearance, occurred. A picture of steatohepatitis with sever fibrosis and cirrhosis was found. A caloric restriction with physical activity as well as the use of metformin were recommended. In the following 6 years the patient develops a decompensated cirrhosis and was referred to a Transplant Unit. This case suggests that non alcoholic steatohepatitis and cirrhosis can develop in patients with metabolic syndrome, especially when other cofactors of liver damage, as alcohol, may be superimposed. In non alcoholic steatohepatitis liver biopsy is mandatory to stage the disease. Moreover it is always recommended when different causes may concur to liver damage in the same patient.

  16. Leucocyte specific antinuclear antibodies. Preparation and fluorescence study. (United States)

    Lampert, I A; Evans, D J; Chesterton, C J


    Tissue and species specific anti-nuclear sera detectable by immunofluorescence were produced by immunising rabbits with D.N.A.-non-histone protein extract from human leucocytes. This is a potentially valuable method for the morphological study of differentiation.

  17. Pre-treatment antinuclear antibody positivity, therapeutic efficacy and persistence of biologics in rheumatoid arthritis

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    Codreanu Cătălin


    Full Text Available Introduction: Rheumatoid factor (RF and anti-citrullinated protein antibodies (ACPA are poor prognostic factors in rheumatoid arthritis (RA. The therapeutic implication of antinuclear antibody (ANA positivity in RA is still debated. The study aims to evaluate ANA positivity as a prognostic factor for the therapeutic response to biologics in RA.

  18. Anti-Nuclear Antibodies in Daily Clinical Practice: Prevalence in Primary, Secondary, and Tertiary Care

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    Thomas Y. Avery


    Full Text Available For the diagnosis of systemic autoimmune rheumatic diseases (SARD, patients are screened for anti-nuclear antibodies (ANA. ANA, as assessed by indirect immunofluorescence (IIF, have a poor specificity. This hampers interpretation of positive results in clinical settings with low pretest probability of SARD. We hypothesized that the utility of positive ANA IIF results increases from primary to tertiary care. We retrospectively determined ANA, anti-ENA, and anti-dsDNA antibody prevalence in patient cohorts from primary (n=1453, secondary (n=1621, and tertiary (n=1168 care settings. Results reveal that from primary care to tertiary care, ANA prevalence increases (6.2, 10.8, and 16.0%, resp.. Moreover, in primary care low titres (70% versus 51% and 52% in secondary and tertiary care, resp. are more frequent and anti-ENA/dsDNA reactivities are less prevalent (21% versus 39% in secondary care. Typically, in tertiary care the prevalence of anti-ENA/dsDNA reactivities (21% is lower than expected. From this descriptive study we conclude that positive ANA IIF results are more prone to false interpretation in clinical settings with low pretest probabilities for SARD, as in primary care. Whether alternative approaches, that is, immunoadsorption of anti-DFS70 antibodies or implementation of anti-ENA screen assays, perform better, needs to be determined.

  19. Comparison of enzyme linked immunosorbent assay (ELISA with indirect immunofluorescence for detection of anti-nuclear antibody

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    G.L.S. Sumanth Kumar


    Full Text Available Background: Detection of antinuclear antibody (ANA is used as one of the diagnostic criteria for autoimmune rheumatic diseases (ARD. Both indirect immunofluorescence (IIF and enzyme linked immunosorbant assay (ELISA methods are used for this purpose. However, there are lack of data comparing these two tests from India. Methods: We prospectively studed 294 patients clinically suspected to be having ARD between April 2012 and September 2013. They were tested for ANA by IIF and ELISA methods. Representative samples positive by both the tests were processed again by a line immunoassay test to detect the specific antinuclear antibodies. Considering the IIF results as the ‘gold standard’, the utility of ELISA for ANA detection was analyzed. Results: Of the 294 samples processed, 181 (61.5% were from female patients. By IIF 30% of samples in males and 40.3% sample in females tested positive. We found ELISA to have a poor sensitivity (45.8% but good specificity (99.5%. The positive predictive value for ELISA were 98% and negative predictive value 76.2% respectively. Forty four samples positive by both IIF and ELISA were tested by Western blot to detect individual autoantibodies. Of these, only 24 samples showed the presence of one or more bands, while the remaining 20 (45.4% were negative by line immunoassay. In our study anti-nuclear ribonucleoprotein/Smith was the most common ANA detected. Conclusions: The poor sensitivity raises concerns regarding the practice of initial screening for ANA by ELISA

  20. Antinuclear antibodies are not increased in the early phase of Borrelia infection.

    NARCIS (Netherlands)

    Spiewak, R.W.; Stojek, NM; Chmielewska-Badora, J


    In the literature, there are case reports suggesting that Borrelia burgdorferi infection may induce autoimmune diseases dependent on antinuclear antibodies (ANA). The present study was undertaken in order to verify this possibility in a prospective manner. The study group comprised 78 consecutive pa

  1. Correlation of antinuclear antibody immunofluorescence patterns with immune profile using line immunoassay in the Indian scenario

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    Sebastian Wendy


    Full Text Available Background: Immunity status, individual response to disease and types of antibodies produced are well known to vary from person to person, place to place and probably from population to population. A broad spectrum of specific auto antibodies that have so far been associated with specific rheumatic diseases, as noted in Western literature, has been well taken as a reference standard all over the world. There is neither research work nor any data correlating the auto antibodies and their antinuclear antibody (ANA patterns with the immunoprofile in the Indian population to date. Aims: To understand a definite association between ANA patterns and specific antibodies in the serum in the Indian study population and to document similarities / differences with the West. Settings and Design: This prospective and retrospective double blind study was undertaken on the South Indian population referred for ANA testing by Indirect Immunofluorescence method and by immunoline methods. Materials and Methods: Serum samples of patients from a random South Indian population who sought medical help for rheumatic disease were subjected for ANA testing by indirect immunofluorescence (IIF method and line immunoassay during the study period of 27 months. Serum samples were processed in dilution of 1:100 using HEp - 2010 / liver biochip (Monkey (EUROIMMUN AG. The serum samples which were further processed for line immunoassay were treated in 1:100 dilution on nylon strips coated with recombinant and purified antigens as discrete lines with plastic backing (EUROIMMUN AG coated with antigens nRNP / Sm, Sm, SSA, Ro-52, SSB, Scl-70, PM-Scl, PCNA, Jo-1, CENP-B, dsDNA, nucleosomes, histones, ribosomal protein-P, anti-mitochondrial antibodies (AMA-M2 along with a control band. The analysis was done by comparing the intensity of the reaction with positive control line by image analysis. Results: The antinuclear antibody indirect immunofluorescence (ANA - IIF patterns obtained

  2. Hydralazine Induced Lupus Syndrome Presenting with Recurrent Pericardial Effusion and a Negative Antinuclear Antibody


    Praneet Iyer; Ahmed Dirweesh; Ritika Zijoo


    Drug induced lupus erythematosus (DIL or DILE) is an autoimmune disorder caused by chronic use of certain drugs. We report a unique case of hydralazine induced lupus syndrome (HILS) with a negative antinuclear antibody in a female patient who was on hydralazine for a period of 1.5–2 years and developed recurrent pericardial effusion as a result of it. Initially her condition was managed with a pericardial window. The recurrence of a massive pericardial effusion necessitated a right hemiperica...

  3. Prevalencia de anticuerpos anti envoltura nuclear y sus isotipos en sueros positivos para anticuerpos antinucleares Prevalence of antinuclear envelope antibodies and their isotypes in sera positive for antinuclear antibodies

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    Miriam Arcavi


    Full Text Available Los anticuerpos antinucleares detectados por inmunofluorescencia indirecta en células HEp-2 presentan una gran variedad de imágenes, entre ellas el patrón de envoltura nuclear que suele ser un hallazgo poco frecuente. Se procesaron 2594 sueros en los cuales se detectó un 37.6% de anticuerpos antinucleares. La prevalencia de anticuerpos anti-envoltura nuclear (ANEA fue del 1.2% presentando una alta asociación con hepatopatías autoinmunes (83% y baja con lupus eritematoso sistémico. En los 21 sueros de los pacientes que presentaron ANEA no se detectaron anticuerpos anti-ADNn hallándose 28.6% de anticuerpos anti-músculo liso y 19% de anticuerpos anti-mitocondriales. El corte triple de tejido de rata mostró ser un sustrato menos sensible que HEp-2 para la detección de ANEA. Al utilizar conjugados dirigidos contra diferentes isotipos de anticuerpos para la detección de ANEA, se encontró: 90.5% de IgG, 66.6% de IgA y 9.5% de IgM. Dos de los pacientes presentaron ANEA-IgA a altos títulos (³1:160 en ausencia de ANEA-IgG. En este trabajo se destaca la importancia de realizar pruebas complementarias que detecten anticuerpos anti-músculo liso, anti-mitocondriales y anti-ADNn, para orientar el diagnóstico clínico de los pacientes que presentan ANEA. Además, sostiene la postura de utilizar como conjugado para IFI-HEp2 anticuerpos anti-inmunoglobulinas totales en lugar de anti-IgG hasta tanto se clarifique el rol que juegan los anticuerpos IgA en estas enfermedades autoimunes.Antinuclear antibodies detected in HEp-2 cells by indirect immunofluorescence assay display a great variety of images, including the nuclear envelope pattern. This is quite a less frequent finding. Two thousand five hundred and ninety-four sera were processed, and 37.6% of ANA were detected. The prevalence of anti-nuclear envelope antibodies (ANEA was of 1.2%, with a high association with autoimmune liver diseases (83% and a low association with systemic lupus

  4. Evaluation of antinuclear antibodies by indirect immunofluorescence and line immunoassay methods': four years' data from Turkey. (United States)

    Sener, Asli Gamze; Afsar, Ilhan; Demirci, Mustafa


    The presence of antinuclear antibodies (ANAs), directed against intracellular antigens, is a hallmark of systemic autoimmune rheumatic diseases. The indirect immunofluorescence (IIF) assay is among the most commonly used routine methods for ANA detection as the screening test. The objective of the study was to evaluate ANA patterns in a 4-year period retrospectively. All 19 996 serum samples that were sent to the Laboratory of Medical Microbiology of the tertiary Hospital by any hospital department between 1 January 2009 and 1 January 2013 with a request to test for ANA, anti-ENA or both were included in the study. Of these samples, 4375 (21.9%) were ANA-IIF-positive and 15621 (78.1%) were ANA-IIF-negative. The presented ANA-positive samples consisted of 2392 (54.67%) homogenous, 818 (18.70%) speckled, 396 (9.05%) centromere, 242 (5.53%) nucleolar, 213 (4.87%) nuclear dots, 178 (4.07%) cytoplasmic (except for actin and golgi), 24 (0.55%) actin, 9 (0.21%) golgi, 53 (1.21%) nuclear membrane and 50 (1.14%) mixed pattern. Totally 7800 samples were examined by LIA. Of these samples, 3440 were positive and 4307 were negative with IIF and LIA. In addition, 22 samples were detected as IIF-positive but LIA-negative, whereas the rest 31 samples were IIF-negative but LIA-positive. ANA patterns in 22 IIF-positive samples were homogenous (9), speckled (5), golgi (4), cytoplasmic (3) and nucleolar (1). SSA/Ro-52, SSB/La and Scl-70 positivity were detected in 31 IIF-negative/LIA-positive samples by LIA. The present study comes forward with its overall scope, which covers 4-year data obtained in tertiary hospital located in the western part of Turkey.

  5. Pathogenic effects of maternal antinuclear antibodies during pregnancy in women with lupus

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    Rafael Herrera-Esparza


    Full Text Available Lupus is an autoimmune disease that primarily affects young women of childbearing age. Fertility rates in lupus patients depend on various factors, including disease activity, nephritis, and the presence of antiphospholipid antibodies; however, after lupus patients become pregnant, different factors may affect the course of pregnancy, such as the production of autoantibodies, pre-existing renal disease, and eclampsia, among others. The placenta is a temporary hemochorial organ that prevents immunological conflict due to exposure to alloantigens at the maternal-fetal interface; placental regulatory T cells play a major role in maternal-fetal tolerance. Typically, significant amounts of maternal IgG class antibodies cross the placenta and enter the fetal circulation. This transition depends on the distribution of Fc receptors along the syncytiotrophoblast. The production of antinuclear antibodies (ANA is a hallmark of lupus, and these autoantibodies can form immune complexes that are typically trapped in the placenta during gestation. However, the entry of ANA into the fetal circulation depends on the IgG-ANA concentration and the FcR placental density. Maternal antinuclear antibodies with anti-Ro or anti-La specificity might be pathogenic to the fetus if transfused by the placental pathway and could induce neonatal pathologies, such as neonatal lupus and congenital heart block. Here, we review the experimental and clinical data supporting a pathogenic role for maternal autoantibodies transmitted to the fetus

  6. Hydralazine Induced Lupus Syndrome Presenting with Recurrent Pericardial Effusion and a Negative Antinuclear Antibody

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    Praneet Iyer


    Full Text Available Drug induced lupus erythematosus (DIL or DILE is an autoimmune disorder caused by chronic use of certain drugs. We report a unique case of hydralazine induced lupus syndrome (HILS with a negative antinuclear antibody in a female patient who was on hydralazine for a period of 1.5–2 years and developed recurrent pericardial effusion as a result of it. Initially her condition was managed with a pericardial window. The recurrence of a massive pericardial effusion necessitated a right hemipericardiectomy. After hydralazine was stopped, she never had any further episodes of pericardial effusion or tamponade.

  7. Detection of antinuclear and antilaminin antibodies in autistic children who received thimerosal-containing vaccines. (United States)

    Singh, Vijendra K; Rivas, Wyatt H


    Autism, a neurodevelopmental disorder, may involve autoimmune pathogenesis. Since mercury is potentially a risk factor for autoimmunity, we conducted a study of mercury-induced antinuclear and antilaminin antibodies in autistic and normal children who had been pre-administered with thimerosal-containing vaccines. Laboratory analysis by different immunoassays showed that the serum level of these two autoimmune markers did not significantly differ between autistic and normal children. This finding suggests that the mercury as in thimerosal-containing vaccines is likely not related to autoimmune phenomenon in autism.

  8. Hydralazine Induced Lupus Syndrome Presenting with Recurrent Pericardial Effusion and a Negative Antinuclear Antibody (United States)

    Iyer, Praneet; Zijoo, Ritika


    Drug induced lupus erythematosus (DIL or DILE) is an autoimmune disorder caused by chronic use of certain drugs. We report a unique case of hydralazine induced lupus syndrome (HILS) with a negative antinuclear antibody in a female patient who was on hydralazine for a period of 1.5–2 years and developed recurrent pericardial effusion as a result of it. Initially her condition was managed with a pericardial window. The recurrence of a massive pericardial effusion necessitated a right hemipericardiectomy. After hydralazine was stopped, she never had any further episodes of pericardial effusion or tamponade. PMID:28194293

  9. Cost-effective detection of non-antidouble-stranded DNA antinuclear antibody specificities in daily clinical practice

    NARCIS (Netherlands)

    Vos, PAJM; Bast, EJEG; Derksen, RHWM


    Objectives. To compare the utility of indirect immunofluorescence for the detection of antinuclear antibodies (ANA-IIF) and a fully automated test (ELiA Symphony (TM)) that detects antibodies against a mixture of nuclear and cytoplasmic antigens (ENA), to select sera that should be tested for non-an

  10. Complement-fixing properties of antinuclear antibodies distinguish drug-induced lupus from systemic lupus erythematosus. (United States)

    Rubin, R L; Teodorescu, M; Beutner, E H; Plunkett, R W


    The immunofluorescence antinuclear antibody (ANA) test has been widely used to monitor autoimmune disease, but its value for diagnostic purposes is compromised by low specificity and high prevalence in disease-free individuals. The capacity of autoantibodies to fix serum complement proteins when bound to antigen is an important effector function because this property is associated with acute and chronic inflammatory processes. The current study evaluates the complement-fixing properties of antinuclear antibodies (CANA) in three well-defined and clinically-related patient groups: systemic lupus erythematosus (SLE), drug-induced lupus (DIL) and drug-induced autoimmunity (DIA). Of 20 patients diagnosed with SLE, 90% displayed complement-fixing ANA while this feature was present in only two of 18 patients with DIL and no patients with DIA without associated disease even though the mean ANA titres were similar among these patient groups. CANA was significantly correlated with anti-Sm activity. Because SLE but not DIL or DIA can be a life-threatening disease associated with complement consumption in vivo, these results demonstrate that measurement of CANA is a diagnostically useful tool and may have immunopathologic implications.

  11. Antinuclear antibodies in rheumatic disease: a proposal for a function-based classification. (United States)

    Pisetsky, D S


    Antinuclear antibodies (ANAs) are a diverse group of autoantibodies that bind macromolecular components of the cell nucleus. While some ANAs occur in normal individuals, others are expressed almost exclusively in patients with rheumatic disease and serve as markers for diagnosis and prognosis. Despite the clinical associations of ANAs, the relationship of these antibodies to specific disease manifestations is often unknown because the target antigens are intracellular molecules that are ubiquitously expressed. In systemic lupus erythematosus, the role of ANAs in disease manifestations is better understood, especially for antibodies to DNA and related nucleosomal antigens. These antibodies can promote nephritis by the formation of immune complexes that are deposited in the kidney. In addition, anti-DNA, along with antibodies to RNA-binding proteins such as anti-Sm, can induce non-specific immune abnormalities based on the induction of type interferon 1 by plasmacytoid dendritic cells. Despite ANA expression in rheumatic disease, studies in animal models of inflammation and tissue injury indicate that antibodies to certain nuclear molecules such as HMGB1 have protective effects. Together, these considerations suggest a function-based classification of ANAs based on their expression in normal and autoimmune individuals as well as their capacity to induce or attenuate immunological disturbances. This classification provides a framework to elucidate the serological features of rheumatic disease and the often uncertain relationship between ANA expression and disease manifestations.

  12. Human peripheral blood monocytes display surface antigens recognized by monoclonal antinuclear antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Holers, V.M.; Kotzin, B.L.


    The authors used monoclonal anti-nuclear autoantibodies and indirect immunofluorescence to examine normal human peripheral blood mononuclear leukocytes for the presence of cell surface nuclear antigens. Only one monoclonal anti-histone antibody (MH-2) was found to bind to freshly isolated PBL, staining approximately 10% of large cells. However, after cells were placed into culture for 16-24 h, a high percentage (up to 60%) of large-sized cells were recognized by an anti-DNA (BWD-1) and several different antihistone monoclonal antibodies (BWH-1, MH-1, and MH-2). These antibodies recognize separate antigenic determinants on chromatin and histones extracted from chromatin. The histone antigen-positive cells were viable, and the monoclonal antibodies could be shown to be binding to the cell surface and not to the nucleus. Using monoclonal antibodies specific for monocytes and T cells, and complement-mediated cytotoxicity, the cells bearing histone antigens were shown to be primarily monocytes. The appearance of histone and DNA antigen-positive cells was nearly completely inhibited by the addition of low concentrations of cycloheximide at initiation of the cultures. In contrast, little effect on the percentage of positive cells was detected if cells were exposed to high doses of gamma irradiation before culture. These data further support the existence of cell surface nuclear antigens on selected cell subsets, which may provide insight into the immunopathogenesis of systemic lupus erythematosus and related autoimmune diseases.

  13. International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies

    NARCIS (Netherlands)

    Agmon-Levin, Nancy; Damoiseaux, Jan; Kallenberg, Cees; Sack, Ulrich; Witte, Torsten; Herold, Manfred; Bossuyt, Xavier; Musset, Lucille; Cervera, Ricard; Plaza-Lopez, Aresio; Dias, Carlos; Sousa, Maria Jose; Radice, Antonella; Eriksson, Catharina; Hultgren, Olof; Viander, Markku; Khamashta, Munther; Regenass, Stephan; Coelho Andrade, Luis Eduardo; Wiik, Allan; Tincani, Angela; Ronnelid, Johan; Bloch, Donald B.; Fritzler, Marvin J.; Chan, Edward K. L.; Garcia-De la Torre, I.; Konstantinov, Konstantin N.; Lahita, Robert; Wilson, Merlin; Vainio, Olli; Fabien, Nicole; Sinico, Renato Alberto; Meroni, Pierluigi; Shoenfeld, Yehuda


    Anti-nuclear antibodies (ANA) are fundamental for the diagnosis of autoimmune diseases, and have been determined by indirect immunofluorescence assay (IIFA) for decades. As the demand for ANA testing increased, alternative techniques were developed challenging the classic IIFA. These alternative pla

  14. Pneumatosis cystoides intestinalis in patients with antinuclear antibody negative systemic lupus erythematosus and dermatomyositis: report of two cases

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Soo Yeon; Cho, On Koo; Koh, Byung Hee; Kim, Yong Soo; Song, Soon Young [College of Medicine, Hanyang University, Seoul (Korea, Republic of)


    Pneumatosis cystoides intestinalis (PCI) occurring in association with collagen vascular disease is an unusual combination that presents with intramural gas in the gastrointestinal tract. We report two cases of PCI, one with antinuclear antibody (ANA) negative SLE and the other with dermatomyositis, with a review of the relevant literature.

  15. A Comparison of Anti-Nuclear Antibody Quantification Using Automated Enzyme Immunoassays and Immunofluorescence Assays

    DEFF Research Database (Denmark)

    Baronaite, Renata; Engelhart, Merete; Mørk Hansen, Troels;


    Anti-nuclear antibodies (ANA) have traditionally been evaluated using indirect fluorescence assays (IFA) with HEp-2 cells. Quantitative immunoassays (EIA) have replaced the use of HEp-2 cells in some laboratories. Here, we evaluated ANA in 400 consecutive and unselected routinely referred patients...... patients were compared. The majority of the results were the same between the two methods (n = 325, 84%); however, 8% (n = 30) yielded equivocal results (equivocal-negative and equivocal-positive) and 8% (n = 31) yielded divergent results (positive-negative). The results showed fairly good agreement......, with Cohen's kappa value of 0.30 (95% confidence interval (CI) = 0.14-0.46), which decreased to 0.23 (95% CI = 0.06-0.40) when the results for dsDNA were omitted. The EIA method was less reliable for assessing nuclear and speckled reactivity patterns, whereas the IFA method presented difficulties detecting...

  16. Antinuclear antibody-negative, drug-induced lupus caused by lisinopril. (United States)

    Carter, J D; Valeriano-Marcet, J; Kanik, K S; Vasey, F B


    The clinical symptoms of drug-induced lupus (DIL) are similar to those of idiopathic systemic lupus erythematosus. The literature indicates that in patients with DIL, sera generally contain antinuclear antibodies (ANAs); however, ANA-negative DIL has been reported. The list of medications implicated as etiologic agents in DIL continues to grow. This list includes two different types of angiotensin-converting enzyme inhibitors--captopril and enalapril. We report the first case of DIL caused by lisinopril. Our patient had negative results on ANA testing and had histone antibodies (IgG anti-[H2A-H2B]-DNA) mirroring the disease course. Antibodies to the (H2A-H2B)-DNA complex are seen in more than 90% of patients with active DIL, excluding those with DIL due to hydralazine. Thus, it is important to recognize the clinical significance of IgG anti-(H2A-H2B)-DNA antibodies and that negative ANA test results do not preclude the diagnosis of DIL.

  17. A case of anti-nuclear matrix protein 2 antibody positive myopathy associated with lung cancer. (United States)

    Ohta, Shin; Unoda, Ki-Ichi; Nakajima, Hideto; Ikeda, Soichiro; Hamaguchi, Yasuhito; Kimura, Fumiharu


    Myositis-specific autoantibodies (MSAs) are associated with myositis. Anti-nuclear matrix protein 2 (NXP-2) antibody was recently identified as a major MSA and was observed mostly in juvenile dermatomyositis. We report the case of a 44-year-old man who presented with myopathy with anti-NXP-2 antibody and large cell carcinoma of the lung. He was hospitalized because of myalgia and edema of limbs. Neurological examination revealed mild proximal-dominant weakness in all four extremities, and laboratory studies showed elevated creatine kinase level (6,432 IU/l). Needle electromyography showed myogenic patterns. MRI of the lower limbs demonstrated inflammatory lesions in the thighs. Biopsied specimen from the left quadriceps femoris muscle showed mild mononuclear inflammatory infiltrate surrounding muscle fibres but no fiber necrosis. He was diagnosed with myopathy based on neurological examinations and clinical symptoms. His chest X-ray and CT showed tumor shadow on the right upper lung field, but CT didn't indicate the findings of interstitial lung disease. This was surgically removed, and a histological diagnosis of non-small cell lung cancer was suspected. He was also treated with definitive chemoradiotherapy before and after operation. His symptoms of myopathy promptly remitted with the preoperative chemotherapy. His serum analysis was positive for the anti-NXP-2. Further investigation and experience of MSAs are necessary to evaluate the therapeutic strategy against cancer-associated myopathy/myositis.

  18. Vulvovaginal-gingival Lichen Planus: Association with Lichen Planopilaris and Stratified Epithelium-specific Antinuclear Antibodies. (United States)

    Olszewska, Malgorzata; Banka-Wrona, Agnieszka; Skrok, Anna; Rakowska, Adriana; Górska, Renata; Solomon, Lynn W; Rudnicka, Lidia


    Vulvovaginal-gingival lichen planus (VVG-LP) consists of a triad of symptoms: vulval, vaginal and gingival lichen planus lesions. The aim of this study was to analyse the prevalence of lesions in various anatomical locations in patients with VVG-LP. The study included 126 consecutive patients with lichen planus. Sixteen (12.7%) patients fulfilled the criteria of VVG-LP. In 12/16 (75%) patients with VVG-LP scalp lesions were also observed. Stratified epithelium-specific antinuclear antibodies (SES-ANA) and anti-ΔNp.3α antibodies were detected in 10/16 (75%) patients with VVG-LP and in 15/110 (13.6%) patients with other forms of lichen planus (p lichen planopilaris. The new entity may be termed "vulvovaginal-gingival-pilar lichen planus" and our study indicates that SES-ANA is a marker of this type of lichen planus with extensive, severe and refractory-to-therapy involvement of the mucous membranes, skin and scalp.

  19. Evaluation of the Canadian Rheumatology Association Choosing Wisely recommendation concerning anti-nuclear antibody (ANA) testing. (United States)

    Ferrari, Robert


    The objective of this study was to evaluate the Canadian Rheumatology Association Choosing Wisely recommendation concerning anti-nuclear antibody (ANA) testing. Patients with joint pain/stiffness/swelling were assessed to determine if ANA testing was indicated. An a priori threshold was set before ANA testing would be considered. Those who did not have ANA testing ordered were followed for 1 year to determine if any of them went on to have a diagnosis of systemic lupus erythematosus (SLE) or other connective tissue disease. A parallel study was conducted with a similar a priori threshold for the use of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) antibody testing in the diagnosis of rheumatoid arthritis (RA), and again, patients were followed for 1 year. A total of 866 subjects were examined, 509 females (58.8 %) and 357 males (41.2 %). The mean age of the group was 47.5 ± 16.8 years. The mean duration of symptoms was 12.0 ± 5.6 weeks. Of the 866 subjects, 68 met an a priori threshold for ordering ANA, RF, and anti-CCP testing. Of these 68, there was a newly diagnosed case of SLE, 4 newly diagnosed cases of RA, and 3 cases of polymyalgia rheumatica. The remaining 798 subjects were followed for approximately 1 year and none developed evidence of SLE, RA, or other connective tissue disease. In the evaluation of non-specific musculoskeletal symptoms, setting an a priori threshold for ordering serology in keeping with the spirit of the Canadian Rheumatology Association Choosing Wisely recommendation for antibody testing results in a very low risk of missing a case of systemic lupus erythematosus or rheumatoid arthritis.

  20. Usefulness of pleural effusion antinuclear antibodies in the diagnosis of lupus pleuritis. (United States)

    Toworakul, C; Kasitanon, N; Sukitawut, W; Wichinun, R; Louthrenoo, W


    We performed this study to determine sensitivity and specificity of pleural effusion antinuclear antibodies (ANA) at a titer of ≥1 : 160, and the ratio of pleural effusion to serum ANA of ≥1, to distinguish between pleural fluid from lupus pleuritis and other causes. A prospective study of 54 patients with pleural effusion (12 lupus pleuritis, seven parapneumonic effusion, 26 malignancy-associated pleural effusions, nine transudative effusions) was performed. ANA at a titer of ≥1 : 160 were found in 11 of 12 lupus pleuritis samples, and in four of 42 pleural effusions from non-systemic lupus erythematosus (SLE) patients. The pleural effusion ANA at a titer of ≥1 : 160 gave a sensitivity of 91.67% for lupus pleuritis, with a specificity of 83.33% when compared with all other pleural effusions, 90.91% when compared with exudative effusion (parapneumonic effusion and malignancy-associated effusion) and 55.56% when compared with the transudative pleural effusion group. Using the ratio of pleural effusion to serum ANA of ≥1, the sensitivity and the specificity decreased to 75.00% and 78.57%, respectively. This study provides further evidence that the pleural effusion ANA at a titer of ≥1 : 160 is a sensitive and specific diagnostic biomarker for lupus pleuritis in patients with lupus. However, pleural effusion ANA can occasionally be found in other conditions.

  1. Antinuclear antibody testing in pleural fluid for the diagnosis of lupus pleuritis. (United States)

    Porcel, J M; Ordi-Ros, J; Esquerda, A; Vives, M; Madroñero, A B; Bielsa, S; Vilardell-Tarrés, M; Light, R W


    We sought to determine whether measuring antinuclear antibodies (ANA) and their specificities [dsDNA, extractable nuclear antigens (ENA)] on pleural fluid may contribute to the differential diagnosis of pleural effusions. ANA were tested by indirect immunofluorescence on Hep-2 cells in the pleural fluid of 266 patients with effusions of different etiologies, including 15 lupus pleuritis. The cutoff value for diagnostic use was set at 1:160. Pleural fluid analysis of specific autoantibodies, such as anti-dsDNA and anti-ENA, was also performed if a positive ANA test was obtained. All patients with lupus pleurisy and 16 of 251 (6.4%) patients with pleural effusions secondary to other causes were ANA positive. Fifty-six percent of the positive ANAs in non-lupus pleural fluids were due to neoplasms. The pleural fluid ANA titers were low (pleuritis from non-lupus etiologies, the absence of pleural fluid anti-dsDNA or anti-ENA favored the latter. ANAs in pleural fluid provided no additional diagnostic information beyond that obtained by the measurement in serum and, therefore, these tests need not be routinely performed on pleural fluid samples. However, in patients with SLE and a pleural effusion of uncertain etiology, lack of ANAs or specific autoantibodies in pleural fluid argues against the diagnosis of lupus pleuritis.

  2. Choosing wisely: Review and commentary on anti-nuclear antibody (ANA) testing. (United States)

    Fritzler, Marvin J


    Choosing Wisely®: Next Steps in Improving Healthcare Value is an initiative of the American Board of Internal Medicine (ABIM) Foundation. The driving forces for the Choosing Wisely (CW) campaign include rising and unstainable health care expenditures and evidence that there is lack of fiscal stewardship of health care resources. The American College of Rheumatology and the Canadian Rheumatology Association published their top five Choosing Wisely recommendations, the first of which pertained to antinuclear antibodies (ANA) and ANA subserology testing. Concerns about the wasteful use of these tests prompted an analysis of the expenditures attributable to ANA testing as a proportion of total health care expenditures and based on a financial model was in the range of 0.00125%. It is suggested that if the sole use of ANA testing is to add evidence to support a diagnosis when the pre-test probability is high, then the ANA test has limited clinical value. Accordingly, the goal of ANA testing needs to be reconsidered and expanded beyond an approach to simply confirming a diagnosis with 'intention to treat' to a goal of case finding of 'pre- or early disease' with an 'intent to prevent' disease. This an area where more significant inroads can be made in preventing end organ disease and thereby reducing health care expenditures HCE. One CW recommendation that bears emphasizing is that, with a few possible exceptions, repeat ANA or ANA subserology testing has little clinical value in monitoring disease activity or predicting a flare.

  3. Comparison of an enzyme immunoassay to an indirect fluorescent immunoassay for the detection of antinuclear antibodies. (United States)

    Reisner, B S; DiBlasi, J; Goel, N


    The standard method for detecting antinuclear antibodies (ANAs) is by immunofluorescence assay (IFA), a method that is labor intensive and subjective. In an attempt to overcome these limitations, several commercial enzyme immunoassays (EIAs) have been developed. We report the results of our evaluation of the ANA Microplate EIA (Sanofi Diagnostics Pasteur, Chaska, MN). For the evaluation, 808 serum samples were tested by EIA and IFA; 52 specimens were positive by both assays, 561 were negative by both assays, 91 were positive by EIA only, and 3 were positive by IFA only. Borderline results (not positive or negative) were obtained for 101 specimens, which were excluded when calculating the sensitivity, specificity, and positive and negative predictive values of this assay, which were 94.6%, 86.0%, 36.4%, and 99.5%, respectively. Because of its high negative predictive value, this assay can be used reliably to detect ANA-negative samples; however, the low positive predictive value indicates that EIA-positive specimens should be retested by an IFA to determine the final result.

  4. Antinuclear, Cytoskeletal, Antineuronal Antibodies in the Serum Samples of Children with Tic Disorders and Obsessive Compulsive Disorders

    Directory of Open Access Journals (Sweden)

    Işık Görker


    Full Text Available streptococcus infections in the development of tic and obsessive compulsive disorders (OCD is controversial. The autoimmune hypothesis states that during infection, formation of autoantibodies leads to an autoimmune disorder, which in turn results in movement disorders, tic disorders and/or OCD. In order to test this hypothesis, we assayed these antibodies in children and adolescents diagnosed with tic disorders and/or OCD.Material and Methods: Children and adolescents who were diagnosed with either tic disorders or OCD according to DSM-IV criteria (n=28, were compared with healthy controls (n=15 having similar age and gender characteristics. Regardless of a streptococcus infection history, serum samples of all patients and controls underwent antinuclear, cytoskeletal, and antineuronal antibody assay using indirect immunofluorescence.Results: The rates of antinuclear antibody positivity were 21% and 20% in the patient and control groups respectively (p>0.05. Antineuronal antibody was positive in 2 (7% of 28 patients versus in 1 (6% of 15 controls (p>0.05.Conclusion: These results suggest that such antibodies may not be involved in the pathogenesis of tic disorders/OCD.

  5. Evaluation of Anti-Nuclear antibody test results in clinical practice

    Directory of Open Access Journals (Sweden)

    Nevreste Çelikbilek


    Full Text Available Objective: Aim of this study is to evaluate anti-nuclear antibody (ANA test results obtained between 2009 and 2011. Methods: Of a totally 5068 cases tested for ANA by indirect immunofluorescence method (IIFA, randomly chosen 982 ANA-positive cases were reviewed in terms of gender, level and pattern of fluorescence, anti-dsDNA (anti-double stranded DNA and anti-extractable nuclear antigen (ENA profile. Anti-dsDNA levels and anti-ENA profiles were determined by enzyme linked immune assay (ELISA and immune-blotting (IB, respectively. Results: Sex distribution of ANA positive patients was determined as 756 (77% females and 226 (23% males. Fifty per cent of the cases were from rheumatology department, 20% from gastroenterology and 30% from other units. Fluorescence levels were considered borderline or weak positive in 62.6% of the samples. The most frequent patterns were homogeneous (23%, speckled (22%, homogeneous-speckled (15.5% and nucleolar (13.5%. Anti-dsDNA were studied in 759 ANA positive patients and 66 (8.7% samples were found positive, being 44 of them (68.8% with homogeneous pattern and the rest with speckled, nucleolar, nuclear dots, centromeric or midbody patterns. Totally 131 (31.6% of 414 samples studied for anti-ENA profile were found positive. The first four frequent profiles were SSA (34.4%, SSA-SSB (16.8%, Scl70 (16% and Sm/RNP (9.2%. Conclusion: Our results are similar with the current related literature. It is known that autoantibodies can be detectable before clinical symptoms being apparent, especially in SLE. Therefore, borderline or weak fluorescence levels should also be reported and the patients having them should be followed-up carefully. J Microbiol Infect Dis 2015;5(2: 63-68

  6. A positive anti-nuclear antibody result does not contraindicate living kidney donation. (United States)

    Lim, C C; Goh, A


    Autoantibodies may precede the clinical onset of disease in up to two-thirds of patients with systemic lupus erythematosus (SLE). There is thus concern that antinuclear antibody (ANA) positivity in living kidney donors may eventuate in development of lupus nephritis post-uninephrectomy. Post-uninephrectomy, we routinely examine living kidney donors at 6 weeks, 6 months, 12 months, and annually thereafter. We performed a retrospective review of living kidney donors who had undergone uninephrectomy between July 1, 1999, and December 1, 2008. Data were collected for pre- and post-uninephrectomy renal function, proteinuria, hematuria, ANA, and anti-double stranded deoxyribonucleic acid (anti-dsDNA) antibodies. Among 68 living donor uninephrectomies performed during the study period, we excluded patients with no pre- uninephrectomy ANA or defaulted postoperative follow-up (n = 2). Twelve (18.2%) living donors were ANA-positive (M:F = 4:8) with a median titer of 1:100 (range, 1:100-1:800); 1 was transiently anti-dsDNA-positive. There were no significant differences between ANA-positive and ANA-negative donors in baseline demographics or pre-uninephrectomy creatinine clearance (median 108.0, range 79-179 mL/min vs. 106, range 58-172 mL/min, P = NS). Only 1 ANA-positive patient displayed transient microscopic hematuria (8 red blood cells); none had proteinuria pre-nephrectomy. Median follow-up was 63.5 (range, 21.2-139.3) months. At last clinic review, none of the ANA-positive donors had developed clinical symptoms or signs of SLE. Comparing ANA-positive and ANA-negative live donors at 5 years post-uninephrectomy, there were no significant differences in creatinine clearance (median 86.5, range 67-112 mL/min vs. 85.0, range 35-154 mL/min, P = NS), total urine protein (median 0, range 0-0.196 g/24 h vs 0, range 0-0.190 g/24 h, P = NS), or presence of hematuria (16.7% vs 33.3%, P = NS). Isolated ANA positivity does not contraindicate living kidney donation. Up to 5 years

  7. Analysis of 15 patients with systemic lupus erythematosus manifesting with negative immunofluorescence anti-nuclear antibodies after treatment. (United States)

    Song, L-J; Ding, F; Liu, H-X; Shu, Q; Yu, X; Li, J; Li, X-F


    The purpose of this study was to investigate the clinical and laboratorial characteristics of patients with systemic lupus erythematosus (SLE) manifesting with negative immunofluorescence anti-nuclear antibodies (IFANA) after treatment for the better understanding of negative conversion of IFANA. Demographic characteristics, clinical and laboratory data of hospitalized SLE patients between March 2006 and May 2011 were retrospectively reviewed. Fifteen cases with negative IFANA were identified in 960 patients. All of the 15 patients were severe, 11 patients manifested with nephritic range proteinuria and hypoalbuminemia, 8 patients were complicated with severe infection and all of the patients had been treated with glucocorticoid and immunosuppressant. Anti-ENA antibodies were positive in 4 of 15 patients. Eight patients died after average 1-year follow-up. Collectively, negative IFANA is mainly attributed to nephritic-range proteinuria; and large-dose glucocorticoid, immunosuppressant and severe infection are also important factors for negative IFANA. Antinuclear antibody can be detected in some SLE patients with negative IFANA by changing the detection method and titer. Negative conversion of IFANA often indicates unfavorable prognosis for severe patients.

  8. [Seroprevalence of antinuclear antibodies in blood donors in the Yaqui Valley]. (United States)

    Serrano-Osuna, Ricardo; López-López, Rocío Milagro; Brito-Zurita, Olga Rosa; Sabag-Ruiz, Enrique; Pérez-Fernández, Héctor; Ornelas-Aguirre, José Manuel


    Antecedentes: los anticuerpos antinucleares son inmunoglobulinas que reconocen componentes celulares nucleares y citoplasmáticos autólogos. En personas sanas no se relacionan con alguna enfermedad autoinmune; sin embargo, pueden estar vinculados con un fenotipo inmunológico de riesgo que no ha sido suficientemente estudiado. Objetivo: examinar la existencia de anticuerpos antinucleares en el suero de donadores de sangre. Material y métodos: estudio transversal y analítico en búsqueda de anticuerpos antinucleares en suero mediante inmunofluorescencia indirecta en células HEp-2. La prevalencia y patrón de expresión se contrastaron con la edad, el género y los antecedentes de enfermedad reumática o tiroidea. Resultados: se estudiaron 379 donadores de sangre con límites de edad entre 18 y 65 años. La prevalencia de anticuerpos antinucleares en la población estudiada fue de 13%. La mayoría de los sujetos positivos tenía entre 21 y 40 años de edad. El género masculino expresó mayor proporción de positividad (11%) en comparación con las mujeres (2%). De la misma forma, 82% de los hombres tenía títulos bajos (1:80) y en 66% eran de tipo nucleolar (RM = 10.66 [1.83 a 62.18]; p = 0.007). Conclusiones: en individuos sanos, la existencia de anticuerpos antinucleares a títulos bajos puede no significar un estado de autoinmunidad; sin embargo, podría ser el reflejo de una exposición a factores ambientales que no han sido lo suficientemente estudiados. Es necesario realizar nuevos estudios en población sana que permitan explicar la asociación entre estos anticuerpos y los factores tóxicoambientales, así como sus efectos en la salud.

  9. Gemcitabine-induced hemolytic uremic syndrome mimicking scleroderma renal crisis presenting with Raynaud's phenomenon, positive antinuclear antibodies and hypertensive emergency. (United States)

    Yamada, Yuichiro; Suzuki, Keisuke; Nobata, Hironobu; Kawai, Hirohisa; Wakamatsu, Ryo; Miura, Naoto; Banno, Shogo; Imai, Hirokazu


    A 58-year-old woman who received gemcitabine for advanced gallbladder cancer developed an impaired renal function, thrombocytopenia, Raynaud's phenomenon, digital ischemic changes, a high antinuclear antibody titer and hypertensive emergency that mimicked a scleroderma renal crisis. A kidney biopsy specimen demonstrated onion-skin lesions in the arterioles and small arteries along with ischemic changes in the glomeruli, compatible with a diagnosis of hypertensive emergency (malignant hypertension). The intravenous administration of a calcium channel blocker, the oral administration of an angiotensin-converting enzyme inhibitor and angiotensin II receptor blocker and the transfusion of fresh frozen plasma were effective for treating the thrombocytopenia and progressive kidney dysfunction. Gemcitabine induces hemolytic uremic syndrome with accelerated hypertension and Raynaud's phenomenon, mimicking scleroderma renal crisis.

  10. Review for the generalist: The antinuclear antibody test in children - When to use it and what to do with a positive titer

    Directory of Open Access Journals (Sweden)

    Sailer-Hoeck Michaela


    Full Text Available Abstract The antinuclear antibody test (ANA is a much overused test in pediatrics. The ANA does have a role in serologic testing but it should be a very limited one. It is often ordered as a screening test for rheumatic illnesses in a primary care setting. However, since it has low specificity and sensitivity for most rheumatic and musculoskeletal illnesses in children, it should not be ordered as a screening test for non-specific complaints such as musculoskeletal pain. It should only be used as a diagnostic test for children with probable Systemic Lupus Erythematosus (SLE or Mixed Connective Tissue Disease, (MCTD and other possible overlap-like illnesses. Such children should have developed definite signs and symptoms of a disease before the ANA is ordered. This review presents data supporting these conclusions and a review of the ANA literature in adults and children. By limiting ANA testing, primary care providers can avoid needless venipuncture pain, unnecessary referrals, extra medical expenses, and most importantly, significant parental anxieties. It is best not to do the ANA test in most children but if it ordered and is positive in a low titer (

  11. RBC Antibody Screen (United States)

    ... test also may be used to help diagnose autoimmune-related hemolytic anemia in conjunction with a DAT. This condition may be caused when a person produces antibodies against his or her own RBC antigens. This can happen with some autoimmune disorders , such as lupus , with diseases such as ...

  12. The effect of TNFalpha blockade on the antinuclear antibody profile in patients with chronic arthritis: biological and clinical implications. (United States)

    De Rycke, L; Baeten, D; Kruithof, E; Van den Bosch, F; Veys, E M; De Keyser, F


    Since the first proof of efficacy of TNFalpha blockade, both the number of patients treated worldwide and the number of indications for treatment with TNFalpha blockers have grown steadily. Surprisingly, the profound immunomodulation induced by anti-TNFalpha therapy is associated with a relatively low incidence of immune-related complications such as lupus-like syndromes and demyelinating disease. This contrasts sharply with the prominent induction of autoantibodies such as antinuclear antibodies (ANA) and anti-dsDNA antibodies during TNFalpha blockade. Although this phenomenon has been recognized for several years, the clinical and biological implications are not yet fully understood. In this review, recent studies analysing the effect of TNFalpha blockade (infliximab and etanercept) on the ANA profile in autoimmune arthritis will be discussed. Taken together, these reports indicate that the prominent ANA and anti-dsDNA autoantibody response is 1) not a pure class effect of TNFalpha blockers, 2) independent of the disease background, 3) largely restricted to the induction of short-term IgM anti-dsDNA antibodies, and 4) not associated with other serological or clinically relevant signs of lupus. Nevertheless, a careful follow-up of patients treated with TNFalpha blockers remains mandatory, including monitoring for lupus-like characteristics.

  13. The effects of affinity-purified anti-DNA antibodies from patients with systemic lupus erythematosus on the fluorescent antinuclear antibody assay using HEp-2 cells. (United States)

    Suzuki, Kimihiro; Kawamura, Masahide; Mineo, Midori; Shinohara, Tadashi; Kataharada, Koji; Okada, Makoto; Takada, Kunio; Miyawaki, Shoji; Ohsuzu, Fumitaka


    The aim of this study was to clarify the effects of anti-dsDNA antibodies on the titer and the nuclear staining pattern(s) in a fluorescent antinuclear antibody (FANA) assay using HEp-2 cells. Anti-dsDNA derived from 14 patients with systemic lupus erythematosus (SLE) was individually affinity-purified. The anti-dsDNA titer of the purified anti-dsDNA solution was measured by radioimmunoassay (RIA) or by enzyme-linked immunosorbent assay (ELISA). In the FANA assay, the anti-dsDNA solution was diluted in a stepwise manner and its titer was expressed by the endpoint dilution. The nuclear staining pattern in the anti-dsDNA solution was examined at the 1:5 and 1:20 dilutions and at the endpoint dilution. The anti-dsDNA titers of the affinity-purified anti-dsDNA solution were high enough (13 to 126 IU/ml) to be measured by RIA. However, the antinuclear antibody (ANA) titers of this solution were relatively low: 1:20 to 1:320. In the study of nuclear staining the peripheral pattern was observed in nine of the 14 cases at a 1:5 dilution. However, at the endpoint dilution, all cases exhibited the homogeneous pattern. These findings indicate that in the FANA assay using HEp-2 cells, 1) although serum samples show high anti-dsDNA titers by RIA or by ELISA, the antibodies' direct contribution to ANA titers is limited, and 2) when samples reveal a homogeneous staining pattern at the endpoint dilution, this suggests the presence of anti-dsDNA.

  14. Apolipoprotein E-knockout mice show increased titers of serum anti-nuclear and anti-dsDNA antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yuehai [Cardiovascular Department, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Huang, Ziyang, E-mail: [Cardiovascular Department, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Lu, Huixia [Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University, Qilu Hospital, Jinan, Shandong 250012 (China); Lin, Huili; Wang, Zhenhua [Cardiovascular Department, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Chen, Xiaoqing [Department of Rheumatism and Immunology, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Ouyang, Qiufang [Cardiovascular Department, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Tang, Mengxiong; Hao, Panpan [Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University, Qilu Hospital, Jinan, Shandong 250012 (China); Ni, Jingqin [Cardiovascular Department, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Xu, Dongming [Department of Rheumatism and Immunology, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); Zhang, Mingxiang; Zhang, Qunye [Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University, Qilu Hospital, Jinan, Shandong 250012 (China); Lin, Ling [Department of Rheumatism and Immunology, Second Clinical Medical College, Fujian Medical University, Quanzhou, Fujian 362000 (China); and others


    Highlights: Black-Right-Pointing-Pointer Titers of ANA and anti-dsDNA antibodies were higher in ApoE{sup -/-} than C57B6/L mice. Black-Right-Pointing-Pointer Spleen was greater and splenocyte apoptosis lower in ApoE{sup -/-} than B6 mice. Black-Right-Pointing-Pointer Level of TLR4 was lower in spleen tissue of ApoE{sup -/-} than B6 mice. Black-Right-Pointing-Pointer The TLR4 pathway may participate in maintaining the balance of splenocyte apoptosis. Black-Right-Pointing-Pointer The TLR4 pathway may participate in antibody production in spleen tissue. -- Abstract: Apolipoprotein E-knockout (ApoE{sup -/-}) mice, atherosclerosis-prone mice, show an autoimmune response, but the pathogenesis is not fully understood. We investigated the pathogenesis in female and male ApoE{sup -/-} mice. The spleens of all ApoE{sup -/-} and C57BL/6 (B6) mice were weighed. The serum IgG level and titers of anti-nuclear antibody (ANA) and anti-double-stranded DNA (anti-dsDNA) antibody were assayed by ELISA. Apoptosis of spleen tissue was evaluated by TUNEL. TLR4 level in spleen tissue was tested by immunohistochemistry and Western blot analysis. Levels of MyD88, p38, phosphorylated p38 (pp38), interferon regulatory factor 3 (IRF3) and Bcl-2-associated X protein (Bax) in spleen tissue were detected by Western blot analysis. We also survey the changes of serum autoantibodies, spleen weight, splenocyte apoptosis and the expressions of TLR4, MyD88, pp38, IRF3 and Bax in spleen tissue in male ApoE{sup -/-} mice after 4 weeks of lipopolysaccharide (LPS), Toll-like receptor 4 ligand, administration. ApoE{sup -/-} mice showed splenomegaly and significantly increased serum level of IgG and titers of ANA and anti-dsDNA antibody as compared with B6 mice. Splenocyte apoptosis and the expression of TLR4, MyD88, pp38, IRF3 and Bax in spleen tissue were significantly lower in ApoE{sup -/-} than B6 mice. The expression of TLR4, MyD88, IRF3, pp38, and Bax differed by sex in ApoE{sup -/-} spleen tissue. The

  15. Simultaneous identification of various antinuclear antibodies using an automated multiparameter line immunoassay system. (United States)

    López-Longo, F J; Rodríguez-Mahou, M; Escalona-Monge, M; González, C M; Monteagudo, I; Carreño-Pérez, L


    The objective was to determine the sensitivity and specificity of an automated multiparameter line immunoassay system compared with other techniques for the identification of autoantibodies in rheumatic diseases. We studied sera from 90 patients. Anti-U1RNP, anti-Sm, anti-Ro/SS-A, anti-La/SS-B, anti-Jo 1 and anti-Scl 70 antibodies were identified by counterimmunoelectrophoresis (CIE) techniques, enzyme-linked immunosorbent assay (ELISA), immunoblotting (IB) using extracts of rabbit thymus and human placenta, and an automated multiparameter line immunoassay system (INNO-LIA ANA UPDATE K-1090) that detects nine different antibodies simultaneously (anti-U1RNP, anti-Sm, anti-Ro/SS-A, anti-La/SS-B, anti-Scl 70, anti-Jo 1, anticentromere, antihistone, and antiribosomal P protein). The line immunoassay system equaled or surpassed the other techniques in the identification of anti-Sm, anti-La/SS-B, anti-Jo 1 and anti-Scl 70 antibodies (sensitivity 100%, specificity 94-100%) and was similarly effective in the case of anti-U1RNP (sensitivity 87.5%, specificity 93.9%) and anti-Ro/SS-A (sensitivity 91.4%, specificity 87.2%) antibodies. In addition, this technique detected more 52 and 60 kD anti-Ro/SS-A sera than IB. Nine antibodies can be detected with this method at a cost of 25.38 Euros per serum sample. In five hours, 19 sera can be studied. The approximate cost of detecting these nine antibodies with an automated ELISA system would be 28.93 Euros, which allows 10 sera to be studied in four hours. In conclusion, the automated multiparameter line immunoassay system is a valid method for the detection of autoantibodies in rheumatic diseases. Its most notable advantages are automated simultaneous detection of several autoantibodies in the same serum and its lower cost compared with ELISA techniques.

  16. Anti-nuclear antibodies positive serum from systemic lupus erythematosus patients promotes cardiovascular manifestations and the presence of human antibody in the brain

    Directory of Open Access Journals (Sweden)

    Marie Kelly-Worden


    Full Text Available Background: Systemic lupus erythematosus (SLE is characterized by the presence of anti-nuclear antibodies (ANAs in the serum of patients. These antibodies may cross over into the brain resulting in the development of neuropsychiatric symptoms and result in abnormal pathology in other organs such as the heart and kidneys. Objective: The objective of this study was to determine if SLE pathology could be detected in the hearts and brains of rats injected with positive human ANA serum. Materials and Methods: Lewis rats (n = 31 were selected for this study due to documented research already performed with this strain in the investigation of serum sickness, encephalitis and autoimmune related carditis. Rats were injected once a week with either ANA positive or negative control serum or saline. Hearts were examined for initial signs of heart disease including the presence of lipid deposits, vegetation, increased ventricular thickness and a change in heart weight. Brains were examined for the presence of human antibody and necrotic lesions. Animals were observed for outward signs of neuropathy as well. Blood samples were taken in order to determine final circulating concentrations of IgG and monitor histamine levels. Results: Animals injected with ANA were significantly higher for lipid deposits in the heart and an increased ventricular thickness was noted. One animal even displayed Libman-Sacks endocarditis. Brains were positive for the presence of human IgG and diffuse internal lesions occurred in 80% of the ANA positive serum injected animals examined. Blood histamine levels were not significantly different, but actually lower than controls by the end of the experiment. Conclusion: Since human antibodies were detected in the brain, further studies will have to identify which antibody cross reactions are occurring within the brain, examine cell infiltration as well as characterize the antibodies associated with more destructive consequences such as

  17. Phenotypic screening: the future of antibody discovery. (United States)

    Gonzalez-Munoz, Andrea L; Minter, Ralph R; Rust, Steven J


    Most antibody therapeutics have been isolated from high throughput target-based screening. However, as the number of validated targets diminishes and the target space becomes increasingly competitive, alternative strategies, such as phenotypic screening, are gaining momentum. Here, we review successful phenotypic screens, including those used to isolate antibodies against cancer and infectious agents. We also consider exciting advances in the expression and phenotypic screening of antibody repertoires in single cell autocrine systems. As technologies continue to develop, we believe that antibody phenotypic screening will increase further in popularity and has the potential to provide the next generation of therapeutic antibodies.

  18. Prevalence of antinuclear and anti-liver-kidney-microsome type-1 antibodies in patients with chronic hepatitis C in China

    Institute of Scientific and Technical Information of China (English)

    BAI Li; FENG Zhen-ru; LU Hai-ying; LI Wen-gang; YU Min; XU Xiao-yuan


    Background Hepatitis C virus (HCV) infection may induce autoimmune response and autoantibodies can be detected in chronic hepatitis C (CHC) patients. However, the reported positive rate of autoantibodies in CHC patients in China varies considerably. In this study, we investigated the prevalence of antinuclear antibodies (ANA) and anti-liver-kidney-microsome type 1 autoantibodies (anti-LKM-1) in a large cohort of CHC patients, and analyzed the factors related to the presence of the autoantibodies. Methods A total of 360 CHC patients were enrolled in this study. Serum ANA and anti-LKM-1 were detected by indirect immunofluorescence and enzyme-linked immunosorbent assay, respectively. Clinical analysis was performed to disclose the related factors to autoantibody production. Results The prevalence of ANA and anti-LKM-1 in CHC patients was 12.5% (45/360) and 2.5% (9/360), respectively. Women had a higher prevalence than men (18.9% vs 11.4%, P=0.046). Patients with positive autoantibodies had lower HCV RNA levels (1.2x107 copies/L vs 7.2x107 copies/L, P <0.05). Positive ANA was associated with higher serum globulin (P<0.05). Stratified analysis showed that there were no significant differences in age, HCV genotype, disease course, clinical stage, prevalence of cirrhosis and interferon therapy between autoantibody-positive and-negative subgroups. Conclusion Autoantibodies can be induced in the course of CHC, and some CHC patients can even develop autoimmune hepatitis.

  19. Aging-dependent decline of IL-10 producing B cells coincides with production of antinuclear antibodies but not rheumatoid factors. (United States)

    van der Geest, Kornelis S M; Lorencetti, Pedro G; Abdulahad, Wayel H; Horst, Gerda; Huitema, Minke; Roozendaal, Caroline; Kroesen, Bart-Jan; Brouwer, Elisabeth; Boots, Annemieke M H


    Aging is associated with development of autoimmunity. Loss of B cell tolerance in the elderly is suggested by an increased prevalence of anti-nuclear antibodies (ANAs) and rheumatoid factors (RFs). Accumulating evidence indicates that B cells also impact autoimmunity via secretion of cytokines. So far, few studies have directly assessed the effect of aging on the latter B cell function. Here, we determined if and how human aging influences the production of cytokines by B cells. In a cross-sectional study, we found that absolute numbers of circulating B cells were similar in 31 young (ages 19-39) and 73 old (age ≥ 60) individuals. Numbers of transitional B cells (CD19(+)CD27(-)CD38(High)CD24(High)) were decreased in old individuals, whereas numbers of naive and memory B cell subsets were comparable in young and old individuals. Short-term in vitro stimulation of whole blood samples revealed that numbers of B cells capable of producing TNF-α were similar in young and old individuals. In contrast, B cells capable of IL-10 production were decreased in old subjects. This decline of IL-10(+) B cells was observed in old individuals that were ANA positive, and in those that were negative for both ANAs and RFs. However, IL-10(+) B cells were remarkably well retained in the circulation of old subjects that were RF positive. Thus, pro-inflammatory TNF-α(+) B cells are retained in the elderly, whereas IL-10(+) B cells generally decline. In addition, our findings indicate that IL-10(+) B cells may differentially impact the development of ANAs and RFs in the elderly.

  20. Antinuclear movement

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Young Hee; Im, Jaeg Yeong


    This book is for antinuclear movement. So, this book introduces many articles on nuclear issues of Asia and the pacific area. The titles of articles are the crusades of Reagan by Werner Plaha, contending between super powers in Europe by Alva Reimer Myrdal, claims of resistance by Daniel Ellsberg, nuclear and the Korean Peninsula by Go, Seung Woo, Liberation but of belief of nuclear weapon by Lee, Young Hee and nuclear weapon in Korea by peter Haze.

  1. 抗核抗体核型与抗核抗体谱检测结果对比分析%Comparative analysis of antinuclear antibody karyotypes and antinuclear antibody spectrum

    Institute of Scientific and Technical Information of China (English)

    陈邁琼; 赵迪; 邓晓刚; 任方; 任圣洁; 杨文杰; 李智


    目的:分析本实验室抗核抗体(ANA)与抗核抗体谱(ANAs)17项联合检测的情况,并探讨两者之间的相关性。方法分别用间接免疫荧光法和免疫印迹法检测患者 ANA 和 ANAs,判断其 ANA 核型和 ANAs 结果,并对各荧光核型和各种特异性抗核抗体结果进行统计分析。结果340例 ANA 阳性患者标本中,颗粒型214例,着丝点型51例,胞浆颗粒型28例,核仁型25例,均质型17例,周边型5例,颗粒型是检出的主要核型。ANAs 17项抗体阳性率为82.94%。214例荧光颗粒型标本中,常见的抗体为抗 SSA/Ro52kd(31.78%)、抗 SSA/Ro60k(d30.84%)和抗 SSB/La(19.16%)抗体;17例荧光均质型标本中,常见的抗体为抗核小体(47.06%)和抗组蛋白(41.18%)抗体;抗 CenpB 对着丝点核型的诊断具有较高的特异性,阳性率为92.16%(47/51)。结论不同荧光核型的 ANA 与 ANAs 抗体之间有一定的关系,但并无绝对的规律可寻,两种 ANA 的检测方法,间接免疫荧光法和免疫印迹法联合应用,有助于自身免疫性疾病(AID)的诊断和预后观察。%Objective To analyze the results of the 17-index combined detection of antinuclear antibodies (ANA)karyotypes and antinuclear antibody spectrum (ANAs)in our laboratory and to investigate the correlation between them.Methods ANA and the specific ANAs were detected by the indirect immunofluorescence(IIF)and the immunoblotting respectively.The results of the ANA karyotypes and the specific ANAs were statistically analyzed.Results Results Among 340 samples of ANA positive,214 cases were the speckled pattern,51 cases were the centromere pattern,28 cases were the cytoplasmic pattern,25 cases were the nucleolar pattern,17 samples were the homogeneous pattern and 5 samples were the peripheral pattern,the speckled pattern was the main de-tected pattern.The positive rate of 17-index ANAs was 82.94%.In

  2. Correlation of antinuclear antibody immunofluorescence patterns with immune profile using line immunoassay%抗核抗体核型与条带免疫抗体谱相关性分析

    Institute of Scientific and Technical Information of China (English)

    郭大文; 张英辉; 单娜; 王冰; 孟冬娅


    目的 分析抗核抗体(antinuclear antibody,ANA)与抗核抗体谱(antinuclear antibodies,ANAs)的相关性.方法 2500例疑似或已确诊自身免疫性风湿性疾病患者,采用间接免疫荧光法检测ANA,其中826例同时应用条带免疫分析法行ANAs分析.结果 ANA阳性1 300例,同时行ANAs的826例患者中ANA(+)/ANAs(+) 182例(22.03%),ANA(-)/ANAs(-)529例(64.04%),ANA(+ )/ANAs(-)111例(13.43%),ANA(-)/ANAs(+)4例(0.50%);ANA阳性者中以颗粒型(62.2%)、胞质颗粒型(13.3%)、均质型(9.7%)、核仁型(8.9%)多见,颗粒型多见SSA/Ro52抗体,均质型多见dsDNA抗体,胞质颗粒型多见AMA-M2;ANAs的阳性率随ANA滴度增大而升高;当滴度为1∶800以上时,>20~40岁组ANA阳性率明显高于其他各年龄组(P<0.01).结论 ANA-间接免疫荧光法是良好的筛查实验,ANA核型、滴度与ANAs相结合对自身免疫性风湿性疾病的诊断与鉴别诊断有重要意义.%Objective To investigate the correlation between antinuclear antibody (ANA) patterns and specific autoantibodies in antinuclear antibodies (ANAs). Methods Serum samples from 2 500 patients with suspected or diagnosed autoimmune rheumatic diseases (ARD) were subjected for ANA testing with indirect immunofluorescence (IIF), in which 826 patients were further processed line immunoassay (LIA) for ANAs. Results Among the analyzed samples(?=2 500), 1 300 were positive for ANA. Among the samples submitted both for ANA-IIF and ANAs-LIA testing (n=826), 182 patients (22. 03%) were observed ANA(+)/ ANAs(+), 529 patients (64.04%) were observed ANA(-)/ ANAs(-), 111 patients (13.43%) were observed ANA(+)/ANAs(-) and only 4 patients (0.05%) were observed ANA(-)/ ANAs( + ). Nuclear speckled (62. 2%), cytoplasmic speckled (13. 3%), homogenous (9.1%) and nucleolar (8.9%) pattern were common in 1 300 positive ANA-IIF findings. Anti-SSA/Ro52 was the most frequent antibody with a nuclear speckled pattern, AMA-M2 was the most frequent

  3. Identification of specific antinuclear antibodies in dogs using a line immunoassay and enzyme-linked immunosorbent assay. (United States)

    Bremer, Hanna D; Lattwein, Erik; Renneker, Stefanie; Lilliehöök, Inger; Rönnelid, Johan; Hansson-Hamlin, Helene


    Circulating antinuclear antibodies (ANA) are commonly present in the systemic autoimmune disease Systemic Lupus Erythematosus (SLE) and in other systemic rheumatic diseases, in humans as well as in dogs. The indirect immunofluorescence (IIF)-ANA test is the standard method for detecting ANA. Further testing for specific ANA with immunoblot techniques or ELISAs is routinely performed in humans to aid in the diagnosis and monitoring of disease. Several specific ANA identified in humans have been identified also in suspected canine SLE but, in contrast to humans, investigation of autoantibodies in canine SLE is mainly restricted to the IIF-ANA test. Our aim was to identify both known and novel specific ANA in dogs and to investigate if different IIF-ANA patterns are associated with different specific ANA in dogs. Sera from 240 dogs with suspicion of autoimmune disease (210 IIF-ANA positive (ANA(pos)) and 30 IIF-ANA negative (ANA(neg))) as well as sera from 27 healthy controls were included. The samples were analysed with a line immunoassay, LIA (Euroline ANA Profile 5, Euroimmun, Lübeck, Germany) and four different ELISAs (Euroimmun). The ANA(pos) dogs were divided in two groups depending on the type of IIF-ANA pattern. Of the 210 ANA(pos) samples 68 were classified as ANA homogenous (ANA(H)) and 141 as ANA speckled (ANA(S)), one sample was not possible to classify. Dogs in the ANA(H) group had, compared to the other groups, most frequently high levels of anti-double stranded deoxyribonucleic acid (dsDNA) and anti-nucleosome ANA. Anti-dsDNA antibodies were confirmed in some dogs with the Crithidia luciliae indirect immunofluorescence test (CLIFT). The frequency of ANA(H) dogs with values above those observed in the healthy group was significantly higher compared to ANA(S) dogs for anti-dsDNA, anti-nucleosome, and anti-histone reactivity. Dogs in the ANA(S) group had, compared to the other groups, most frequently high levels of anti-ribonucleoproteins (RNP) and

  4. Antinuclear Antibodies (ANA) (United States)

    ... Pharyngitis, Adenitis Syndrome (Juvenile) Polymyalgia Rheumatica Psoriatic Arthritis Raynaud's Phenomenon Reactive Arthritis Rheumatoid Arthritis Scleroderma Sjogren's Syndrome Spinal Stenosis Spondyloarthritis Systemic Lupus Erythematosus (Juvenile) Takayasu's ...

  5. ANA (Antinuclear Antibody Test) (United States)

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  6. 抗核抗体与抗核抗体谱联合检测在自身免疫性疾病诊断上的应用%Amplification of the combinational detection of antinuclear antibody and antinuclear antibodies spectrum in the diagnosis of autoimmune diseases

    Institute of Scientific and Technical Information of China (English)

    林益强; 曾燕丽; 周建锋; 陈美珺


    目的 探讨抗核抗体(antinuclear antibody,ANA)和抗核抗体谱(antinuclear antibodies spectrum,ANAs)联合检测在自身免疫性疾病(autoimmune diseases,AID)诊断上的应用方案及价值,从而降低现症及潜在AID患者的漏检率.方法 对1231份本院门诊和住院疑似AID患者的血清标本分别采用间接免疫荧光法(indirect immunofluorescence,IIF)和免疫印迹法(immumoblot test,IBT)检测ANA和ANAs,对其中IIF-ANA1∶100(-)/IBT-ANAs(+)标本分别进行1∶10及1∶32稀释后采用IIF检测ANA,最后对上述检测结果 进行统计分析.结果 1231份血清标本中ANA阳性414份,占33.63%,ANAs阳性429份,占34.85%.按不同检测方案进行分组,发现同时检测IIF-ANA与IBT-ANAs的方案对确诊/疑似AID病人检出率最高(43.05%),分别与应用IIF-ANA进行AID的初筛的方案、仅检测IBT-ANAs的方案相比均有统计学意义(x2=23.12,P< 0.05,x2=17.42,P< 0.05).对116份(9.42%)IIF-ANA 1∶100(-)/IBT-ANAs(+)的标本分别进行1∶32及1∶10稀释后再采用IIF检测ANA,发现其中103份标本ANA荧光结果 ≥1∶32,10份标本ANA荧光结果 <1∶32而≥1∶10,3份标本ANA荧光结果 <1∶10;将标本增加1∶32和1∶10稀释度可将AID确诊/疑似病例检出率分别提高8.37%(x2=20.84,P< 0.05)与9.18%(X2=24.70,P<0.05),但二者之间相比则无统计学意义(X2=0.17,P>0.5).结论 IIF-ANA可应用于大范围AID病人初筛以减轻病人经济负担及实验室工作量压力.但是仅检测IIF-ANA或IBT-ANAs均可导致临床上患有AID或潜在的AID病人的漏检.联合检测IIF-ANA和IBT-ANAs,尤其是对临床高度怀疑AID、且ANA荧光结果 ≥1∶32的标本进行IBT-ANAs的检测可显著提高检出率,从而降低现症及潜在AID患者的漏检率.%Autoantibody is an important diagnostic index of autoimmune diseases (AID). In order to investigate the diagnostic value of antinuclear antibody (ANA) and antinuclear antibodies spectrum (ANAs

  7. [Laboratory-based evaluation of "INOVA/QUANTA Lite" to determine antinuclear antibodies (ANA) and autoantibodies to double-stranded DNA, SS-A and SS-B]. (United States)

    Yamauchi, Megumi S; Shingaki, Naohiko; Yamane, Nobuhisa


    We evaluated QUANTA Lite reagent series (INOVA Diagnostics, CA, USA) to determine antinuclear antibodies (ANA) and autoantibodies to double-stranded (ds) DNA, SS-A and SS-B, in parallel with MESACUP (Medical & Biological Laboratories, Nagoya). Overall agreements between two reagents for qualitative interpretation ranged from 77.5% (ANA) to 99.0%(anti-SS-B antibodies). When we compared to the results by indirect fluorescent antibody (IFA) test on HEp-2 cells, QUANTA Lite ANA demonstrated better sensitivity and specificity; 92.2% versus 76.5% in sensitivity and 92.1% versus 86.8% in specificity. Also, determining anti-chromatin antibodies and IFA test onto Chrithidia luciliae demonstrated greater interpretive correlation to detect anti-ds DNA by QUANTA Lite than by MESACUP. All the discrepant sera to which QUANTA Lite SS-A gave positive interpretations were confirmed to contain the antibodies specific to SS-A 52kDa antigen, which is supplemented to QUANTA Lite capture-probes. With these results, we can conclude that QUANTA Lite has superiorities over MESACUP; (1) to detect a variety of autoantibodies consisting of ANA, (2) to have a better correlation with confirmatory tests to detect anti-ds DNA antibodies, (3)to detect additional autoantibodies specific to SS-A 52kDa antigen, and (4) to have an enough compatibility in determining anti-SS-B antibodies.

  8. Production and Screening of Monoclonal Peptide Antibodies. (United States)

    Trier, Nicole Hartwig; Mortensen, Anne; Schiolborg, Annette; Friis, Tina


    Hybridoma technology is a remarkable and indispensable tool for generating high-quality monoclonal antibodies. Hybridoma-derived monoclonal antibodies not only serve as powerful research and diagnostic reagents, but have also emerged as the most rapidly expanding class of therapeutic biologicals. In this chapter, an overview of hybridoma technology and the laboratory procedures used routinely for hybridoma production and antibody screening are presented, including characterization of peptide antibodies.

  9. Comparative analysis of antinuclear antibody spectrum by immunoblotting assays%免疫印迹法检测抗核抗体谱比对分析

    Institute of Scientific and Technical Information of China (English)

    蒋跃根; 应葳; 王峰来; 蔡秀玲; 李安生


    目的 比较抗核抗体谱-12S检测(antinuclear antibody profile,ANA-12S)试剂盒和抗核抗体谱线性免疫分析法检测(antinuclear antibody spectrum linear immunoassay,IMTEC-ANA-LIA)试剂盒对血清抗核抗体谱(antinuclear antibody spectrum, ANAs)检测结果的一致性,从而验证ANA-12S检测ANAs的有效性.方法 收集于该院就诊的间接免疫荧光法(indirect immunofluorescence,IIF)检测抗核抗体(ANA)阳性患者标本144例,其中男21例,女123例;ANA阴性患者标本10例,其中男2例,女8例.用IMTEC-ANA-LIA和ANA-12S分别进行ANAs检测.比较和分析两种方法所得的检测结果.结果 ANA阳性标本中两者检测阳性率均为100%,其中ANA-12S法检测阳性项目占28.8% (497/1 728),IMTEC-ANA-LIA法检测阳性项目占27.7%(479/1 728),差异无统计学意义(x2=0.46,P>0.05),ANA-12S法与IMTEC-ANA-LIA法检测结果总符合率为97.1%,Kappa值为0.929;ANA阴性标本中两者检测的阴性率均为70%(7/10),其中3例系统性红斑狼疮(SLE)疑似患者抗SmD1阳性,阴性项目占97.5%(117/120),总符合率100%,Kappa值为1.结论 ANA-12S与IMTEC-ANA-LIA的检测结果具有较高符合率和准确性;ANA-12S试剂盒可以作为快速、简便、经济的检测方法应用于自身抗体的检测.

  10. Predictive value of antinuclear antibodies in autoimmune diseases classified by clinical criteria: Analytical study in a specialized health institute, one year follow-up (United States)

    Soto, María Elena; Hernández-Becerril, Nidia; Perez-Chiney, Ada Claudia; Hernández-Rizo, Alfredo; Telich-Tarriba, José Eduardo; Juárez-Orozco, Luis Eduardo; Melendez, Gabriela; Bojalil, Rafael


    Introduction: Determination of antinuclear antibodies (ANA) by indirect immunofluorescence (IIF) is usually the initial test for the diagnosis of systemic rheumatic diseases (SRD). Assigning predictive values to positive and negative results of the test is vital because lack of knowledge about ANAs and their usefulness in classification criteria of SRD leads to inappropriate use. Methods: Retrospective study, ANA tests requested by different specialties, correlation to patients' final diagnosis. Results: The prevalence of autoimmune disease was relatively low in our population yielding a low PPV and a high NPV for the ANA test. 40% of the patients had no clinical criteria applied prior to test. Coexistence of two or more autoimmune disorders affects prevalence and predictive values. Conclusion: Application of the test after careful evaluation for clinical criteria remarkably improves the positive likelihood ratio for the diagnosis. PMID:26623249

  11. Anticuerpos antinucleares, imágenes y características obtenidas por inmunofluorescencia: Importancia de los isotipos IgA, IgM e IgG Antinuclear antibodies, patterns and characteristics obtained by immunofluorescence: The importance of the IgA, IgM and IgG isotypes

    Directory of Open Access Journals (Sweden)

    Miriam Arcavi


    Full Text Available La técnica de elección para el screening de anticuerpos antinucleares (ANA es la inmunofluorescencia indirecta que utiliza como sustrato una línea de células epiteliales de carcinoma de laringe humano (IFI-HEp2, y como antisuero, anti-IgG o anti-Ig totales. Los ANA-IgG son los más importantes para el diagnóstico y monitoreo de las enfermedades del tejido conectivo (ETC, mientras los ANA-IgM son de menor relevancia clínica en estos pacientes. Sin embargo, poco se sabe de los ANA-IgA ya que estos Ac han sido menos investigados. El objetivo de este trabajo fue estudiar la prevalencia de los diferentes isotipos de inmunoglobulinas de anticuerpos antinucleares en los pacientes con ETC y evaluar la conveniencia de utilizar conjugados monovalentes o polivalentes. Se procesaron 100 sueros de pacientes con diversas ETC empleando IFI-HEp2, en los cuales se detectó 38% de ANA-IgA (títulos ≥ 1:80 y 12% de ANA-IgM (títulos ≤ 1:160. En 29 casos se detectó IgA en ausencia de IgM, en 3 casos IgM en ausencia de IgA. En todos los casos los ANA-IgG estuvieron presentes. En 6 sueros se observó un cambio de imagen con conjugado anti-IgA y en 3 con conjugado anti-IgM. Debido a la alta prevalencia de ANA-IgA detectada por IFI-HEp2, se destaca la conveniencia de utilizar conjugado anti-Ig totales en lugar de anti-IgG, mientras se desconozca la relevancia de los ANA-IgA en el diagnóstico, pronóstico y seguimiento de las enfermedades reumáticas sistémicas.The indirect immunofluorescence with epitelial cell line from human laryngeal carcinoma as substrate (IIF-HEp2 and anti-IgG or anti-total Ig as antisera, is the technique currently used for the detection of antinuclear antibodies. The most important antibodies for the diagnosis and follow-up of connective tissue diseases (CTD are the IgG-ANA, while the IgM-ANA have no clinical relevance. However the IgA-ANA have not been thoroughly investigated so far. The aim of this work was to study the prevalence

  12. Value of Combined Detection of Antinuclear Antibody,Antinuclear Antibody Profile and Anti-double Stranded DNA Antibody in the Diagnosis of Systemic Lupus Erythematosus%抗核抗体、抗核抗体谱及抗双链DNA抗体的联合检测系统性红斑狼疮的诊断价值

    Institute of Scientific and Technical Information of China (English)

    曾燕坤; 吴杰


    目的 探讨抗核抗体(ANA)、抗核抗体谱(ANAs)及抗双链DNA抗体(抗dsDNA抗体)的联合检测对系统性红斑狼疮(SLE)的诊断价值。方法 选择2012年1月至2014年1月间泉州市某市级医院收治的SLE患者150例作为SLE组,同期收治的其他风湿性疾病患者30例作为疾病对照组,门诊体检健康者30例作为正常对照组。应用间接免疫荧光法检测血清 ANA ,应用免疫印迹法检测血清ANAs和抗dsDNA抗体水平。结果 SLE组ANA、抗dsDNA抗体、抗AHA、抗nRNP、抗AnuA、抗Sm阳性率显著高于疾病对照组和健康对照组( P <0.01)。S L E患者中仅5.33%的患者出现单项抗体阳性,其余94.67%的患者一次性检测出2项或以上抗体阳性。SLE患者治疗后ANA、抗dsDNA抗体、抗AHA、抗Sm和抗SS‐A阳性率均不同程度下降,其中ANA、抗dsDNA抗体、抗AHA好转率分别为23.33%、83.33%和63.33%。结论 SLE患者ANA、ANAs和抗ds‐DNA抗体阳性率较高,联合检测和动态观察 ANA、ANAs和抗 ds‐DNA抗体对SLE诊断和治疗具有一定意义。%Objective] To explore the value of antinuclear antibody(ANA) ,antinuclear antibody profile(ANAs) and anti‐double stranded DNA antibody (anti dsDNA antibody ) in the diagnosis of systemic lupus erythematosus (SLE) .[Methods] Totally 150 patients with SLE in a municipal hospital of Quanzhou city from Jan .2012 to Jan . 2014 were selected as SLE group ,and other 30 patients with rheumatic diseases were selected as disease control group ,and 30 health examination persons were selected as normal control group .Serum ANA was detected by indi‐rect immunofluorescence method .Serum ANAs and anti dsDNA antibody were measured by western blotting .[Re‐sults] The positive rates of serum ANA ,anti dsDNA antibody ,anti‐AHA ,anti‐nRNP ,anti‐AnuA and anti‐Sm in SLE group were significantly higher than those in disease control group and healthy

  13. Simultaneous expression of displayed and secreted antibodies for antibody screen.

    Directory of Open Access Journals (Sweden)

    Yuanping Zhou

    Full Text Available The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS analysis, as well as in conditioned medium in a secreted version for function analysis.

  14. 间接免疫荧光法筛查抗核抗体与特异性抗体检测的相互关系%Relationship between Anti-nuclear Antibodies and the Detection of Specific Anti-nuclear Antibodies

    Institute of Scientific and Technical Information of China (English)

    胡朝军; 李俊; 张道强; 张蜀澜; 李丽君; 董晓娟; 张奉春; 李永哲


    时导致的AID患者漏诊.%Objective To analyze the correlation between the antin-uclear antibody (ANA) results of large samples by indirect immunofluorescence (IIF) screening and specific anti-nuclear antibodies test and study the clinical significance of them and clarify whether they could replace each other in clinical practice.Methods 2 026 cases of consecutive clinical samples for ANA testing were tested by IIP with Hep-2 and line immunoassay (LIA) for the detection of specific ANA antibodies. All the samples were divided into autoimmune diseases ( AID) group, suspected AID group and non-AID group. The relationship between different test results and their clinical significance were analyzed. Results Of the 2 026 cases of specimens, 882 cases (43.53%) were IIF-ANAVLIA-ANAs+ , 266 cases (13. 13%) were IIF-ANAV LIA-ANAs ~ , 507 cases (25.02%) were IIF-ANA" /LIA-ANAs" and 206 cases (10. 17%) were IIF-ANA ~ /LIA-ANAs+. The overall compliance rate of IIF-ANA and LIA-ANA was 68. 56% , the consistency rate was moderate (K=0. All, P <0. 01), and there was significant difference between the results of IIF-ANA and UA-ANA {x2 = 416.21, P < 0.01). The positive rates of anti-Ro-52, anti-SS-A (Sjogren' s syndrome antigen A) , anti-dsDNA (double-stranded DNA) , anti-AMA-M2 (Anti-mitochondrial antibody M2) , anti-Sm (Smith antigen), anti-SS-B (Sjogren's syndrome antigen B), anti-nRNP/Sm (nuclear ribonucleoprotein/Smith antigen) and anti-Jo-1 (Jo-1 antigen) antibodies ranged from 6. 00% to 34.94% in the IIF-ANA"/LIA-ANAs+/1 group. Of the 312 cases of this group, 37. 18% patients (116 cases) were diagnosed as AID, higher than the non-AID group (22. 11 % ) (x2 = 16.97, P < 0. 01). Of the 325 cases of IIF-ANA+ /LIA-ANAs '' * group, 48. 0% of patients (156 cases ) were diagnosed as AID, higher than the non-AID group (22. 15% ) , and the proportion of patients with AID was higher than non-AID in each group of patients with IIF-ANA liter 1:80, 1:160-1 =320 and 2* 1:640 (x2 = 26.96, 7. 89, 19. 42, P < 0.01). Conclusions It

  15. Update on Anti-Saccharomyces cerevisiae antibodies, anti-nuclear associated anti-neutrophil antibodies and antibodies to exocrine pancreas detected by indirect immunofluorescence as biomarkers in chronic inflammatory bowel diseases: Results of a multicenter study

    Institute of Scientific and Technical Information of China (English)

    S Desplat-Jégo; JC Grimaud; M Veyrac; P Chamouard; RL Humbel; C Johanet; A Escande; J Goetz; N Fabien; N Olsson; E Ballot; J Sarles; JJ Baudon


    AIM:Anti-Saccharomyces cerevisiae antibodies (ASCA), anti-nuclear associated anti-neutrophil antibodies (NANA) and antibodies to exocrine pancreas (PAB), are serological tools for discriminating Crohn's disease (CrD) and ulcerative colitis (DC). Like CrD, coeliac disease (CoD) is an inflammatory bowel disease (IBD) associated with (auto) antibodies. Performing a multicenter study we primarily aimed to determine the performance of ASCA, NANA and PAB tests for IBD diagnosis in children and adults, and secondarily to evaluate the prevalence of these markers in CoD.METHODS: Sera of 109 patients with CrD, 78 with UC, 45 with CoD and 50 healthy blood donors were retrospectively included. ASCA, NANA and PAB were detected by indirect immunofluorescence (IIF).RESULTS: ASCA+/NANA- profile displayed a positive predictive value of 94.2% for CrD. Detection of ASCA was correlated with a more severe clinical profile of CrD and treatment of the disease did not influence their serum levels. ASCA positivity was found in 37.9% of active CoD.PAB were found in 36.7% CrD and 13.3% CoD patients and were not correlated with clinical features of CrD, except with an early onset of the disease. Fifteen CrD patients were ASCA negative and PAB positive.CONCLUSION: ASCA and PAB detected by IIF are specific markers for CrD although their presence does not rule out a possible active CoD. The combination of ASCA, NANA and PAB tests improves the sensitivity of immunological markers for CrD. Repeating ASCA, NANA, and PAB testing during the course of CrD has no clinical value.

  16. Mercury exposure, serum antinuclear/antinucleolar antibodies, and serum cytokine levels in mining populations in Amazonian Brazil: a cross-sectional study. (United States)

    Gardner, Renee M; Nyland, Jennifer F; Silva, Ines A; Ventura, Ana Maria; de Souza, Jose Maria; Silbergeld, Ellen K


    Mercury is an immunotoxic substance that has been shown to induce autoimmune disease in rodent models, characterized by lymphoproliferation, overproduction of immunoglobulin (IgG and IgE), and high circulating levels of auto-antibodies directed at antigens located in the nucleus (antinuclear auto-antibodies, or ANA) or the nucleolus (antinucleolar auto-antibodies, or ANoA). We have reported elevated levels of ANA and ANoA in human populations exposed to mercury in artisanal gold mining, though other confounding variables that may also modulate ANA/ANoA levels were not well controlled. The goal of this study is to specifically test whether occupational and environmental conditions (other than mercury exposure) that are associated with artisanal gold mining affect the prevalence of markers of autoimmune dysfunction. We measured ANA, ANoA, and cytokine concentrations in serum and compared results from mercury-exposed artisanal gold miners to those from diamond and emerald miners working under similar conditions and with similar socio-economic status and risks of infectious disease. Mercury-exposed gold miners had higher prevalence of detectable ANA and ANoA and higher titers of ANA and ANoA as compared to diamond and emerald miners with no occupational mercury exposure. Also, mercury-exposed gold miners with detectable ANA or ANoA in serum had significantly higher concentrations of pro-inflammatory cytokines IL-1beta, TNF-alpha, and IFN-gamma in serum as compared to the diamond and emerald miners. This study provides further evidence that mercury exposure may lead to autoimmune dysfunction and systemic inflammation in affected populations.

  17. Evaluation on the detection of anti-nuclear antibody and anti-soluble nuclearantigen antibody for the diagnosis of autoimmune disease%ANA和ENA检测在自身免疫性疾病中的应用评价

    Institute of Scientific and Technical Information of China (English)

    陆慧琦; 李畅; 叶伟民; 杨洁; 何铭珺; 韩焕兴


    To evaluating the sensitivity and correlation on anti-nuclear antibody(ANA) and anti-soluble nuclear antigen antibody (ENA) in the diagnosis of autoimmune disease(AID ), the results of auto-antibody detection in serum were retrospectively analyzed for patients visited at Changzheng hospital from Jan, 2007 to Oct, 2009, among which 224 cases were patients with AID and 325 cases were patients with non-autoimmune diseases. For all these patients, the indirect immunofluorescene(IIF)assay based on the cultured human epidermoid carcinoma cells( Hep-2 cell/liver and immunoblot assay were used to detect ANA and ENA respectively. It was demonstrated that 4 different patterns were formed by using these two kinds of detection methods, i.e. ANA+/ENA+、 ANA-/ENA-、 ANA+/ENA- and ANA-/ENA+ respectively. Positive rate of ANA and ENA (63.4% vs 58. 5%) in AID was significantly higher than that of non-AID (16.9% vs 25.2%). Expect in with MCTD and SLE, there were no correlation between ANA and ENA. The ANA-IIF assay appears to be a time-consuming procedure and difficult to standardize owing to subjective interpretation of results. Acting as a screening experimental method, the sensitivity of ANA is higher than that of ENA. Nevertheless. the combined use of both methods would increase the sensitivity and reliability of testing.%探讨抗核抗体和抗可溶性核抗原抗体的不同检测方法对自身免疫性疾病诊断的指导意义.回顾性分析2007年1月至2009年10月间在长征医院就诊的患者血清自身抗体的检测结果,549位患者,其中自身免疫病患者224例,非自身免疫病325例,所有患者血清同时检测ANA和ENA.以Hep-2细胞/肝组织为基质的间接免疫荧光法检测ANA,免疫印迹法检测ENA.两种检测方法产生4种检出模式:ANA+/ENA+、ANA-/ENA-、ANA+/ENA-和ANA-/ENA+.前两种模式共占检测的62.84%.ANA和ENA在自身免疫病患者中的阳性率(63.4%和58.5%)显著高于非自免病患者(16.9%和25

  18. The clinical significance on the inconsistent results of antinuclear antibody and specific auto antibody determinations%抗核抗体与特异性自身抗体检测结果不一致的临床意义分析

    Institute of Scientific and Technical Information of China (English)

    董晓微; 胡朝军; 张道强; 李丽君; 张蜀澜; 董晓娟; 李永哲


    Objective To compare the inconsistent results of the indirect immunofluorescence ( IIF ) assay screening for antinuclear antibodyv ANA) and linear immunoblot assay( LIA) for antinuclear antibody spectrum ( ANAs ) specific antibody, and evaluate the relationship and clinical significance. Methods The samples were determined for ANA screening by IIF and for ANAs by LIA. The patients with the results of IIF-ANA+ and LIA-ANAs and IIF-ANA' and LIA-ANAs+ were classified into autoimmune disease ( AID ) group and non-AID group. Results In the 216 cases with IIF-ANA" of AID group, The LIA-ANAs + rate was 46. 30% , and the positive rate was higher than that of non-AID group ( 20. 00% , P =0. 00 ). The ratios of LIA-ANAs( + ) and LIA-ANAs( ± ) of AID group were higher than those of non-AID group( P 0. 05 ). However, when the fluorescence titer was 2e 1: 1 280, the difference between 2 groups was statistically significant ( P 0. 05 ). Conclusions In the diagnosis of AID, besides the screening determination by IIF, it is necessary to detect the specific auto antibodies in AID patients in order to avoid the missed detection of AID.%目的 比较间接免疫荧光法(IIF)筛查抗核抗体(ANA)与线性免疫印迹法(LIA)检测抗核抗体谱(ANAs)特异性抗体结果不一致性,分析二者的相互关系及临床意义.方法 采用IIF筛查ANA和LIA检测ANAs特异性抗体,将IIF-ANA阳性/LIA-ANAs阴性和IIF-ANA阴性/LIA-ANAs阳性患者分为自身免疫性疾病(AID)组和非AID组进行比较分析.结果 216例IIF-ANA阴性AID组中,LIA-ANAs阳性率为46.30%,高于非AID组(20.00%,P=0.00);AID组LIA-ANAs(+)、LIA-ANAs(±)的比率均高于非AID组(P 0.05);而荧光滴度为≥1:1280时,2组间的差异有统计学意义(P0.05).结论 在AID的临床诊断中,除进行IIF筛查ANA外,还需根据具体情况进行特异性自身抗体的检测,避免单一方法检测导致AID患者的漏诊.

  19. The correlation study of autoimmunity antinuclear antibody and aging%抗核抗体与衰老的相关性

    Institute of Scientific and Technical Information of China (English)

    杨蓬勃; 谢志贤


    Objective To investigate the influences of age and sex on the expression of serum antinuclear antibody (ANA) and its titer level.Methods The serum ANA in 993 people aged 60 years and over were detected and titer of positive serum were analyzed. They were divided into different groups according to age and sex. Two hundred healthy cases aged less than 60 years were collected as control group. Results The serum ANA positive rate was increased with age and it was significantly higher in people aged 60 years and over than aged less than 60 years (29. 1% vs. 14.0%, x2=5. 074, P=0. 025). The serum ANA positive rates were 24.1%, 30. 5% and 33.6% in people aged 60-69 group, aged 70-79 group and aged above 80 group. Among people aged 60 years and over, the positive rate was 35.8% in female group, which was higher than in male group (22. 4%) (x2= 21. 179, P=0. 05). The titre levels of the positive samples were increased with age. In control group, the samples with 1 : 100 titre was 57.1% among the positive samples, which was higher than in the elderly group (36. 3%). And in control group, the samples with 1 : 3200 titre was 0 among the positive samples, which was lower than in the elderly people group (7.6%). The ANA preliminary screening in people aged 60 years and over showed that the titre level was less than 1 : 320-1: 1000 in female and less than 1 : 100-1: 320 in male. Its specificity could be up to about 90%. Conclusions The serum ANA positive rate in people aged 60 years and over is affected by age and sex. The positive rate is higher in female than in male and it is increased with age. It suggests that 1 : 320 to 1 : 1000 could be taken as ANA preliminary screening critical value in female and 1 : 100 to 1 : 320 in male.%目的 探讨年龄、性别对血清抗核抗体(ANA)表达的影响.方法 对993例老年人进行血清ANA检测,并依年龄及性别不同分组,同时检测200例中青年健康体检者作对照分析.结果 老年组ANA阳性率为29.1%.

  20. 1801例抗核抗体检测结果及其临床分析%Clinical analysis of 1 801 cases detected antinuclear antibodies

    Institute of Scientific and Technical Information of China (English)



    and clinical data in 1 801 cases w ere analyzed .R esults (1 )604 cases (33 .5% ) w ere positive w ith A N A ,of w hich 82 .5% w ere fern ale patients . In patients equalw ith or less than 60 years old , the posi-tive rates w ere significantly higher in fern ale patients than in m ale patients , and the positive rate reached the highest level patients of 36-45 years old .(2) The m ajar ANA fluorescence pattern was antinuclear speckled pattern (43 .7% ) .(3) In patients positive w Mi A N A , the positive rate of specific antibodies w as 47 .8% . T he detection rate of specific antibodies w Mi m ixed pattern w as the highest(57 .8% )and the anticytoplasm ic pattern w as the low est(12 .7% ) (P<0 .01) . Specific antibodies could not be identified in patients positive w ith antinucleolinus , antinuclear m em brane ,anticentriole and anticentrom ere pattern . (4 )66 .2% of A N A -positive patients,yet only 17 .0% of AN A-negative patients, were diagnosed as A ID (P<0 .01) . 73 .6% of them ixed pattern group ,70 .2% of antinuclear pattern group and 36 .9% of the anicy top lam ic pattern group w ere diagnosed as A ID ( P<0 .01). The positive rate of ANA was significantly higher in system ic lupus erythematosus(SLE) group . Positive rates of AN A in groups of SLE , rheum atoid arthritis(R A ) , m ixed connected tissue disease (M C TD ) , system ic scleroderm a(SS ), polym yositis (PM )/ derm atom yositis (DM) and overlap syndrom e were statistical different with patients without A ID (P<0 .05),whereas there was no statistical difference be-tween anky losing spondylitis(AS) and S till disease(ST ILL ) patients and patients without AID . The maJDr ANA pattern in SLE , RA and M CTD patients was antinuclear speckled pattern . In patients with ANA titerat least 1:3 200 , 89 .1% were with A ID , the ratios w ere 72 .2% and 44 .7% in patients w ith A N A titerat least 1:1 000 and 1:320 ,and the ratios w ere statistically differ-entbetween the there groups(P=0 .000) .Conclusion The

  1. Analysis of the results of antinuclear antibody spectrum which antinuclear antibody were negative and assessment of the risks of disease%抗核抗体阴性病例的抗核抗体谱的检测分析与疾病风险的评估

    Institute of Scientific and Technical Information of China (English)

    卢伟; 刘斌剑; 梁艳; 杨再兴


    Objective By analyzing the results of antinuclear antibody spectrum(ANAs)which antinuclear antibody (ANA)were negative,to explore the correlation between them and make clinical assessment of the risks of disease. Methods Make a statistics of the results of serum ANAs cases which ANA were negative and clinical data of 6003 dif-ferent patients came from outpatient and?inpatient from 2011 until May 15,2014.The relationship among the results and their clinical significance were analyzed.Results Of 6003 ANA negative specimens,the positive rate of ANAs was 10.9%,the negative rate of ANAs was 89.1%,the positive rate of anti-SSA antibody was the first (3.9%),the posi-tive rate of anti-Ro-52 antibody was the second(3.6%).Of ANAs positive specimens,the weakly positive rate of anti-Sm,anti-SSB,anti-Scl-70 and anti-His antibodies were higher than the positive of them,the positive rate of anti-Ro-52 antibody was higher than the weakly positive,there were all with statistical significance (P <0.05).The total positive rate of single event’s antibody was the highest(8.7%).Of the weakly positive cases and positive cases of single event’ s antibody,the positive cases of two events’s antibody,the cases of one weak positive and the other positive of mixed e-vents’s antibody,the female positive rate was higher than the male and there was with statistical significance(P <0.05).In the different age group of ≤20 years old,21-49 years old and ≥50 years old,there was statistical significance of ANAs positive rate(6.2%,9.5% 12.6%,χ2 = 21.23,P < 0.05).Of 652 ANAs positive specimens,patients diag-nosed as autoimmune disease(AID)were 104 cases(16.0%).The positive antibodies of ANAs positive specimens were 162 cases,the positive rate of anti-Ro-52 antibody in the AID group was the first (30.9%),the positive rate of anti-SSA antibody was the second(28.4%).Of 5351 ANAs negative specimens,patients diagnosed as AID were 322 cases(6. 0%).The AID patients in ANAs positive specimens were

  2. Prevalence of antinuclear antibodies in 3 groups of healthy individuals: blood donors, hospital personnel, and relatives of patients with autoimmune diseases. (United States)

    Marin, Guadalupe G; Cardiel, Mario H; Cornejo, Horacio; Viveros, Martha E


    Antinuclear antibodies (ANA) are frequently found in healthy populations. To define the prevalence, pattern, and titer of ANA in different groups of the healthy Mexican population, we studied 304 individuals, classified into 3 groups: 104 blood donors, 100 hospital personnel working at The State General Hospital, which included doctors, laboratory technicians, and nurses; and 100 relatives of patient diagnosed either with systemic lupus erythematosus or rheumatoid arthritis, all of them apparently healthy at the time of study. We determined ANA using immunofluorescence microscopy performed on HEp-2 cells. Fluorescence was detected in 165 serum samples (54.3%). The most frequent pattern was the speckled (50.3%). The most frequent dilution was 1:40 (35.4%), followed by 1:80 (13.4%), 1:160 (3.2%), and 1:320 (1.3%).Regarding the results by study group, we found a trend toward higher ANA levels in group 2 (hospital personnel), compared with group 1 (blood donors) and group 3 (relatives of patients), a trend also reflected by the increasing frequency of serum titers of 1:80 and higher (P = 0.074). According to occupation, medical doctors showed a higher incidence of speckled pattern when compared with other occupations (P = 0.022). Medical doctors (n = 75) showed also higher titers of this particular pattern (P = 0.03). In group 3, relatives of patients with systemic lupus erythematosus showed the speckled pattern more frequently than relatives of patients with rheumatoid arthritis, in low titers (P = 0.017). We suggest that ANA tests showing speckled pattern should be at a 1:160 titer or higher to be considered positive; other patterns such as homogeneous, peripheral, or centromeric might be considered positive even at low titers (

  3. 系统性红斑狼疮抗核抗体和抗核抗体谱联合检测及其临床意义%Combined detection of antinuclear antibody and antinuclear antibody spectrum and its clinical significance in systemic lupus erythematosus

    Institute of Scientific and Technical Information of China (English)

    储红颍; 杨桂斌; 王建华; 牛林; 袁国龙


    Objective To explore the diagnostic value and clinical significance of the combined detection of antinuclear antibody (ANA) and anti-nuclear antibody spectrum (ANAs) in systemic lupus erythematosus (SLE) .Methods 110 patients with SLE ,88 patients with other autoimmune diseases (AID) and 50 individuals with healthy physical examination were selected and detected se-rum ANA by using indirect immunofluorescence (IIF);the Western blot was adopted to detect the 15 items of ANAs .Results A-mong 110 cases of SLE ,the positive rates of serum ANA ,anti-ribonucleoprotein /Smith antibody (nRNP/Sm) ,anti-Smith antibody (Sm) ,anti-Sjogren′s syndrome A antibody (SSA) ,anti-Ro-52 antibody (Ro-52) ,anti-Sjogren′s syndrome B antibody (SSB) ,anti-scleroderma 70 antibody (Scl-70) ,anti-PM-Scl antibody (PM-Scl) ,anti-cytoplasmic group acyl-tRNA antibody (J0-1) ,anti-centro-mere antibodies (CENP B) ,anti-proliferative protein antibody (PCNA ) ,anti-double stranded DNA antibody (ds-DNA ) ,anti-nu-cleosome antibody (AnuA) ,anti-histone antibody (AHA) ,anti-ribosomal P protein antibody (ARPA) and anti-mitochondrial anti-body M2 subtype (AMA M2) were 98 .2% ,59 .1 % ,39 .1 % ,71 .8 % ,68 .2 % ,21 .8 % ,2 .7 % ,3 .6 % ,0 .9% ,9 .1% ,5 .5% , 44 .5% ,38 .2 % ,27 .3 % ,38 .2% and 15 .5% respectively ,the positive rate of above 16 kinds of autoantibody in the orther AID were64.8% ,14.8% ,0% ,37.5% ,42.0% ,11.4% ,9.1% ,0% ,5.7% ,9.1% ,1.1% ,2.3% ,1.1% ,1.1% ,5.7% and2.3% respec-tively ;ANA had the highest diagnostic sensitivity (98 .2% ) and low specificity (35 .2% ) for SLE ,the antibodies with higher speci-ficity in diagnosing SLE were anti-Sm antibody (100 .0% ) ,anti-ds-DNA antibody (97 .7% ) ,anti-AnuA antibody (99 .0% ) ,anti-AHA antibody (99 .0% ) ,anti-ARPA antibody (94 .3% ) and anti-PCNA antibody (98 .9% ) and the disease control group (50 .9% ) .In detecting ANA karyotype by IIF ,the maximum was nuclear particle type (43 .5% ) and the disease control group (50 .9% ) ,no

  4. 组蛋白诱导抗核抗体产生及其对肾脏损害%Induction of antinuclear antibodies and systemic lupus erythematosus like syndrome by immunization with histone

    Institute of Scientific and Technical Information of China (English)

    马爱妞; 吴厚生


    目的 寻找系统性红斑狼疮中诱导抗核抗体(ANA)生成的真正的自身核成分免疫原。方法 从ConA活化的脾淋巴细胞中提取组蛋白免疫同系BALB/c小鼠,ELISA方法测定IgG类抗组蛋白、抗dsDNA抗体,免疫荧光法检测抗核抗体核型和免疫复合物在肾小球内的沉积,免疫印迹法测定抗可溶性核抗原抗体,考马斯亮蓝法检测尿蛋白含量,光镜下观察肾脏形态变化,电镜下观察肾小球电子致密物。结果 活性组蛋白诱导IgG类抗组蛋白、抗dsDNA等多种抗核抗体生成,且诱发同系小鼠产生SLE样肾脏病理变化。结论 活性组蛋白是诱发抗核抗体生成、造成肾损害的免疫原之一。%Objective To identify the immunogen capable of inducingproduction of antinuclear antibodies and triggering renal injury. Methods BALB/c mice were immunized with active histone purified from syngeneic spleen cells following Con A activation. The patterns of antinuclear antibodies and immune complexes in glomeruli were observed by immunofluorescence staining. The antibodies against determinants of extractable nuclear antigen (ENA) were demonstrated by immunoblot assay. The dense deposits in glomeruli and kidney injury were observed under electron microscopy and microscopy. Results Active histone induced the production of anti-histone, anti-dsDNA, anti-Ro/SS-A, anti-La/SS-B and anti-J0-1. The nuclear patterns of antinuclear antibodies included homogeneous, speckled, centromeric, peripheral, cytoplasmic, nucleolar patterns, etc. The mice immunized with active histone had the glomerulonephritis and the deposits of IgG immune complex in the glomeruli and in the matrix of the mesangial region and the protein urea. However, the mice immunized with resting histone, which was purified from the syngeneic spleen cells without Con A stimulation failed to do the same. Conclusion Active histone could be one of the immunogens driving the production of anti

  5. Clinical application of antinuclear antibodies in autoimmune diseases%抗核抗体检测在自身免疫性疾病的临床应用

    Institute of Scientific and Technical Information of China (English)

    苏娜; 刘晓玲; 窦存瑞


    Objective To discuss the clinical significance of anti-nuclear antibody (ANA) in treatment of autoimmune diseases (AID). Methods 2055 patients up to the diagnosis standards were selected. The ANA in serum of patients was measured by indirect immunofluorescence. The positive rate, karyotype and titer of ANA were analyzed in the above AID. Results The titer of ANA in AID patients group was 1:80 while 1 :160 in the control group. ANA positive rate of AID patients group was 80%, and its difference with control group had statistical significance (P <0.01). The highest rate was in systemic lupus erythematosus (SLE) patients. Fluorescence patterns of ANA statistics were spot-type based for SLE, drying syndrome, rheumatoid arthritis patients. The homogenous and the nuclear core spots were mainly for autoimmune hepatitis patients. Conclusion The positive rate and titer of ANA are different among various AID patients. More attention should be paid to the titer value of ANA in the ANA test. Because more and more types of ANA fluorescence patterns are found, and different models represent different fluorescent target antigen. In particular, some special fluorescent models play an important role in the diagnosis of AID.%目的 探讨抗核抗体(ANA)在自身免疫性疾病(AID)中的临床意义.方法 选取符合AID诊断标准的患者2055例,采用间接免疫荧光法对患者血清标本进行ANA检测,分析ANA的阳性率、核型及滴度的变化.结果 ANA滴度AID患者以1:160为主,对照组以1:80为主.AID检测患者中,ANA阳性率为80%,与对照组比较有显著性差异(P<0.01),以系统性红斑狼疮的ANA阳性率为最高.ANA荧光模型结果系统性红斑狼疮、干燥综合症、类风湿关节炎患者以斑点型为主,自身免疫性肝炎患者以核均质型及核斑点性为主.结论 不同AID患者检测ANA阳性率和滴度有差异,检测ANA应重视ANA滴度的价值.ANA荧光模型种类较多,不同的荧光模型表示不同的

  6. 检测抗核抗体对系统性红斑狼疮的临床应用%The clinical value of quantitative determination of antinuclear antibody in systemic lupus erythematosus

    Institute of Scientific and Technical Information of China (English)



    目的 探讨检测抗核抗体对系统性红斑狼疮的临床价值.方法 采用回顾性分析的方法,分析我院收治的系统性红斑狼疮患者的临床资料.结果 SLE患者核颗粒型、核均质型、浆颗粒型、浆均质型、核仁型、着丝点型各型阳性率均高于对照组,SLE组总阳性率明显高于对照组,P<0.05.两种狼疮治疗后ANA含量均较治疗前有所降低,但神经性狼疮ANA降低效果明显优于狼疮肾炎患者,P<0.05,差异均有统计学意义.结论 ANA可以作为系统性红斑狼疮患者发生、发展的临床预警指标,值得临床推广检测.%This study is aimed to investigate the clinical value of quantitative determination of antinuclear antibody (ANA) in systemic lupus erythematosus (SLE). We retrospectively studied the clinical records of patients with systemic lupus erythematosus in Hezhou People's Hospital. According to the results, positive rates of speckled patterns, homogeneous patterns, nucleolus pattern and centromere pattern were higher than that of control group, and total positive rate of SLE was significantly higher than that of control group (P<0.05). The antinuclear antibody content of patients with systemic lupus erythematosus was decreased significantly after treatment, while a statistically significant difference was found between neuropsychiatric systemic lupus erythematosus and lupus nephritis group (P<0.05). We conclude that antinuclear antibody can be used as a clinical index for monitoring incidence and progression of systemic lupus erythematosus.

  7. 86例系统性红斑狼疮患者抗核抗体谱检测结果分析%Analysis test results with antinuclear antibody of 86 cases systemic lupus erythematosus patients

    Institute of Scientific and Technical Information of China (English)

    王淑英; 王峰; 王贞美; 张影秋


    目的:探讨抗核抗体谱(ANAs)中12项抗体检测结果对诊断系统性红斑狼疮(systemic lupus erythemato-sus,SLE)的临床应用价值。方法采用线性免疫分析法(line immuno assay LIA)检测86例 SLE 患者(SLE 组)、81例其他风湿性疾病患者(疾病对照组)和90例健康体检者(健康对照组)的血清 ANAs 中12项抗体,采用 SPSS 统计学软件对检测结果分析。结果健康对照组中针对12项抗原的抗体未被检出,SLE 组和疾病对照组中12项抗体有不同程度的阳性被检出,抗双链 DNA 抗体(dsDNA)、抗核小体抗体(Nucleosome)、抗核糖核蛋白粒子抗体(Sm)、抗PO 抗体(P0)、抗组蛋白抗体(Histone)、抗 Ro/SS-A 抗体(52 KD)等7项抗体在 SLE 组的阳性检出率分别为65.12%、53.49%、33.72%、43.02%、47.67%、37.21%、58.14%均明显高于疾病对照组和健康对照组,组间比较差异有统计学意义(P <0.05),疾病对照组与健康对照组间所检测抗体的阳性检出率,比较差异无统计学意义(P >0.05)。SLE 组单项抗体检测与多项抗体联检的阳性率,比较差异有统计学意义(P <0.05),疾病对照组单项抗体检测与多项抗体联检的阳性率,比较差异无统计学意义(P >0.05)。结论ANAs 的12项抗体联检有助于提高 SLE 的阳性检出率,可为疾病的筛查、诊断、分类、病情观察等方面提供有价值的实验室依据,具有较高的临床应用价值。%Objective To investigate the antinuclear antibody (ANAs)in 12 antibody test results for the diagnosis of systemic lupus erythematosus (systemic lupus erythematosus,SLE)value in clinical application.Methods SLE as-say (immuno line LIA)was used to testing the serum ANAs of 12 patients (SLE group),81 patients with other rheu-matic diseases (control group)and 90 healthy controls (healthy control group),and the

  8. Antinuclear human autoantibodies as markers in Nicotiana tabacum pollen tubes

    Directory of Open Access Journals (Sweden)

    C. Poggialini


    Full Text Available In the present paper we report on the use of antinuclear human autoantibodies as specific markers in Nicotiana tabacum pollen tubes. The antibodies have been tested by fluorescence techniques using a confocal laser scanning microscope. All the antibodies showed specifc labelling pattern and the results, although preliminary in nature, could open new perspectives of research.

  9. Analysis of the result of antinuclear antibody and its profile for 1725 patients%1725例血清抗核抗体及抗核抗体谱检测结果分析

    Institute of Scientific and Technical Information of China (English)

    李艳; 孙家祥; 鄂建飞


    目的 探讨抗核抗体(ANA)和ANA谱在自身免疫性疾病(AID)中的临床价值及相符程度.方法 分析1725例标本血清ANA及ANA谱检测结果.ANA和抗双链DNA抗体采用间接免疫荧光法(IIF),ANA谱采用欧蒙印迹法.结果 1725例标本血清ANA阳性共768例,阳性率为44.52%.荧光模式主要为棱颗粒型、核均质型、核仁型等;1725例标本血清ANA谱检测有782例阳性,阳性率为45.33%,大多为抗体二联及以上共同出现阳性,阳性抗体出现较多的是抗SS-A、抗Ro-52、抗SS-B、抗Histone等.结论 ANA和ANA谱联合检测在AI0D诊断中具有互补性,可提高检出率,对AID的诊断分型、治疗、预后判断和病情随访等均有重要的临床价值.%Objective To explore the clinic value and the extent consistent of antinuclear antibodies(ANA) and an-tinuclear antibody profile in the autoimmune diseases! AID). Methods Retrospective analysis of the result of ANA and ANA profile in sera of 1725 patients. ANA and anti-double-stranded DNA antibody were detected by indirect immunoflu-orescence (IIF), ANA profile were detected by Euronline(immunoblot assay). Results 768 ANA positive cases were found out of the 1725 patients, the positive rate being 44. 52%. The main fluorescent type of ANA were nuclear particles pattern, nuclear homogenous pattern, nucleolar pattern. Of all the 1725 samples,782 cases is positive for ANA profile. The positive rate is 45. 33%. Most of the antibodies appear together two or more positive joint, the most frequently occurred antibody was anti-SS-A, anti-Ro-52, anti-SS-B and anti-Histone. Conclusion ANA and ANA profile can supplement each other in diagnosing AID and increase the detection rate, which is of great clinic value to diagnostic classification .treatment, post-prediction judge and disease follow-up of AID.

  10. Comparative analysis of anti-nuclear antibody pattern detection and specific anti-nuclear antibody spectrum detection%抗核抗体核型检测与特异性抗核抗体谱检测的对比分析

    Institute of Scientific and Technical Information of China (English)

    郑金菊; 牟晓峰


    目的:探讨抗核抗体核型与特异性抗核抗体谱的相关性。方法回顾分析本院检验科检测的974例抗核抗体结果,分别用间接免疫荧光法(IIF)检测抗核抗体核型,用条带酶免分析法(LIA)检测特异性抗核抗体谱,分析199例 IIF 与 LIA 同时阳性的结果,比较抗核抗体核型与特异性抗核抗体谱的相关性。结果974例标本中 IIF 阳性249例(阳性率25.6%),LIA 阳性237例(阳性率24.3%),两种方法检测阳性率差异无统计学意义(P >0.05),IIF 与 LIA 单项或两项阳性287例(29.5%),高于IIF 或 LIA 单项检测阳性率,差异有统计学意义(P <0.05)。249例 IIF 阳性中 LIA 阳性199例(79.9%),725例 IIF 阴性中 LIA阴性687例(94.8%)。核颗粒型多见抗 Ro-52抗体,胞浆颗粒型多见抗线粒体 M2抗体,核均质型以抗 dsDNA 抗体、抗核小体抗体多见,着丝点型多见抗着丝点抗体,核仁型多见抗 PM-Scl 抗体。结论IIF-ANA 与 LIA-ANA 有较好的相关性,但也有一定差异,两者联合检测能降低漏检率,对自身免疫性疾病的诊断、病情监测及预后判断有重要意义。%Objective To investigate the correlation between the anti-nuclear antibody(ANA)pattern and the specific ANA spectrum.Methods 974 cases of detected ANA results in our hospital were analyzed retrospectively.The ANA pattern was detec-ted by the indirect immunofluorescence(IIF)and the specific ANA spectrum was tested by the line immunoassay(LIA).199 cases of both simultaneous positive results by IIF and LIA were analyzed and the correlation between ANA patterns and specific ANA spectrum was analyzed.Results Among 974 cases of specimen,249 cases(25.6%)were positive by IIF and 237 cases(24.3%) were positive by LIA,the difference in the positive rate between IIF and LIA had no statistical significance(P >0.05 ).287 cases (29.5%)were positive by single IIF or LIA

  11. Distribution and relevance study of aquaporin-4 antibodies and antinuclear antibodies in patients with optic neuritis%水通道蛋白4抗体及抗核抗体在视神经炎中的分布及相关性分析

    Institute of Scientific and Technical Information of China (English)

    王颖云; 魏世辉


    Objective To observe the seroprevalence of autoimmune antibodies in patients with optic neuritis,mainly including aquaporin-4 antibodies(AQP4-Ab),antinuclear antibodies(ANAs),and analyze the relevance of distribution between AQP4-Ab and ANAs.Methods Ninety-six patients with optic neuritis in hospital from October 2010 to February 2012,all were screened for AQP4-Ab,ANAs and anticardiolipin antibodies,partial for rheumatoid antibodies(APF,AKA,CCP)and HLA-B27,and statistics the positive outcome.Results Among of all patients,the positive rate of AQP4-Ab,ANAs,anticardiolipin antibodies,APF,HLA-B27 were 19.8%(19/96),13.5%(13/96),2.1%(2/96),6.7%(3/45)and 7.1%(1/14),respectively.AQP4-Ab and/or ANAs were positive in 22 patients at a rate of 22.9%(22/96),of which 10 cases showed the two positive simultaneous at a rate of 10.4%(10/96),with a prevalence statistically in distribution of AQP4-Ab and ANAs(P<0.01).Conclusions Minority patients with optic neuritis show positive autoimmune antibodies,including AQP4-Ab,ANAs,anticardiolipin antibodies.The distribution of AQP4-Ab and ANAs exist correlation statistically.%目的 观察水通道蛋白4抗体(AQP4-Ab)及抗核抗体(ANAs)等多种自身免疫抗体在视神经炎中的分布及其相关性.方法 收集2010年10月至2012年2月间在解放军总医院神经眼科病房住院诊治的视神经炎患者96例,全部检测AQP4-Ab、ANAs及抗心磷脂抗体,部分检测类风湿三项(APF、AKA、CCP)及人白细胞抗原-B27(HLA-B27),统计其阳性率,分析AQP4-Ab及ANAs的相关性.结果 观察视神经炎患者中AQP4-Ab、ANAs、抗心磷脂抗体、类风湿三项及HLA-B27阳性率分别为19.8%(19/96)、13.5%(13/96)、2.1%(2/96)、6.7%(3/45)和7.1%(1/14);AQP4-Ab和(或)ANAs阳性者22例,占22.9%(22/96)两者均为阳性者10例,占10.4%(10/96),其分布在统计学差异有统计学意义(P<0.01).结论 少数视神经炎患者表现有AQP4-Ab和(或)ANAs、抗心磷脂抗体等

  12. 抗核抗体谱检测在诊断系统性红斑狼疮中的意义%Clinical Significance of Anti-nuclear Antibodies Detection in the Diagnosis of Systemic Lupus Erythematosus

    Institute of Scientific and Technical Information of China (English)

    蒋素莹; 卢永芳; 谢丹萍; 张金珍


    探讨抗核抗体(ANA)和抗核抗体谱(ANAs)检测对诊断系统性红斑狼疮(SLE)患者的临床意义.用间接免疫荧光法(IIF)和免疫印迹法检测106例SLE组和30名对照组血清ANA及ANAs中的12种抗体.结果表明:SLE组ANA阳性率为86.8%,ANAs中抗ds-DNA、抗Scl-70、抗Jo-1、抗nRNP、抗Sm、抗SS-A、抗SS-B、抗Ro-52、抗CENP-B、抗AnuA、抗AHA和抗核糖体P蛋白的阳性率分别为28.3%、0.9%、0.9%、35.9%、17.0%、39.6%、18.9%、41.5%、9.4%、29.3%、31.1%和9.4%.ANA和ANAs联合检测提高了诊断的敏感性,对SLE的诊断和治疗有重要意义.%Objective To explore the clinical significance of anti-nuclear antibody (ANA) detection in the diagnosis of systemic lupus erythematosus (SLE) patients.Methods The serum ANA and anti-nuclear antibody spectrum (ANAs) of 12 kinds of antibodies in 106 SLE patients and 30 healthy control group were detected by indirect immunofluorescence assay (IIF) and western blot detection.Results The results showed that the positive rate of ANA was 86.8% in SLE patients,and positive rates of ANAs in anti-ds-DNA,anti-Scl-70、antiJo-1 、anti-nRNP,anti-Sm,anti-SS-A,anti-SS-B,anti-Ro-52,anti-CENP-B,anti-AnuA,anti-AHA and the ribosomal P-protien were 28.3%,0.9%,0.9%,35.9%,17.0%,39.6%,18.9%,41.5%,9.4%,29.3%,31.1% and 9.4%,respectively.Conclusion The combined detection of ANA and ANAs could improve the sensitivity and have important significance in the diagnosis and treatment of SLE.

  13. Avaliação clínico-laboratorial de pacientes com síndrome antifosfolípide primária segundo a frequência de anticorpos antinucleares (FAN Hep-2 Clinical and laboratory evaluation of patients with primary antiphospholipid syndrome according to the frequency of antinuclear antibodies (ANA Hep-2

    Directory of Open Access Journals (Sweden)

    Jozélio Freire de Carvalho


    Full Text Available OBJETIVO: Avaliar a frequência de manifestações clínicas e laboratoriais em pacientes com síndrome antifosfolípide primária (SAFP com anticorpos antinucleares positivos (FAN Hep-2+, comparados àqueles com esses anticorpos negativos (FAN Hep-2 -. PACIENTES E MÉTODOS: Estudo transversal em 58 pacientes (82,8% mulheres com SAFP. Foram avaliados os dados demográficos, clínicos, comorbidades, medicações e anticorpos antifosfolípides. RESULTADOS: Dos 58 pacientes incluídos no estudo, vinte (34,5% apresentaram presença de FAN Hep-2. Comparando-se o grupo de pacientes FAN Hep-2+ com aqueles FAN Hep-2 -, verificou-se que ambos os grupos de pacientes com SAFP não diferiram estatisticamente em relação aos dados demográficos, bem como em relação ao tempo de doença. Em relação às manifestações clínicas e laboratoriais, o grupo com FAN Hep-2 + apresentou maior frequência de trombose venosa profunda (85 versus 52,6%, P = 0,04, uma frequência estatística e significativamente maior de anticardiolipina IgG (85 versus 52,6%, P = 0,02 e uma tendência para anticardiolipina IgM (80% versus 52,6%, P = 0,05, bem como maiores medianas desses anticorpos [33 (0-128 versus 20 (0-120 GPL, P = 0,008] e [33 (0-120 versus 18,5 (0-120 MPL, P = 0,009]. Tal diferença não foi observada no que se refere a outras manifestações da SAF, presença de comorbidades, estilo de vida e uso de medicações. CONCLUSÃO: Pacientes com SAFP que apresentam FAN Hep-2+ têm maior frequência de trombose venosa profunda e anticardiolipinas IgG e IgM.OBJECTIVE: To evaluate the frequency of clinical and laboratory manifestations in patients with primary antiphospholipid syndrome (PAPS with positive antinuclear antibodies (ANA Hep-2+ compared to those in whom this antibody is negative (ANA Hep-2-. PATIENTS AND METHODS: This is a transversal study with 58 patients (82.8% females with PAPS. Demographic and clinical data, comorbidities, medications, and

  14. 健康体检人群不同性别抗核抗体随年龄分布规律探讨%Analysis of the distribution of antinuclear antibodies in a healthy population

    Institute of Scientific and Technical Information of China (English)

    郭亚平; 王春光; 刘欣; 刘锦梅; 郭德立; 杨松; 高振庄


    Objective To study the distribution of antinuclear antibodies ( ANAs) in a healthy population and the significance of using ANAs screening test in medical examination .Methods The ANAs were measured by indirect immunofluorescence assay ( IIF) .The Western blot assay was used to detect fif-teen specific antibodies against auto-antigens .Results 3519 out of all 25 110 subjects showed ANAs titers>1∶100 , and among them male and female subjects were respectively accounted for 1143 and 2376 .1489 out of all subjects had ANAs titers >1∶320 , and among them male and female subjects were respectively accounted for 406 and 1083 .The positive rates of ANAs at different titers showed significant differences be -tween male and female subjects .Among subjects with ANAs titers >1∶320 , the number of male subjects showed a steady increase with the age , while the percentage of female subjects reached to two peaks during the periods of puberty and menopause .The fifteen specific antibodies were detected in 659 out of 1489 sub-jects with ANAs titers>1∶320 and anti-Ro-52 (14.2%) accounted for the majority , followed by anti-M2 (12.7%) and anti-SSA (9.6%).Conclusion ANAs can be detected among healthy population of all ages, but their distribution varied with gender and age .ANAs screening test is necessary for medical exami-nation of healthy population , especially for female during period of puberty or menopause .The population with positive ANAs should be followed-up closely and educated for the prevention of autoimmune diseases .%目的:通过分析健康人群抗核抗体( ANA)检测资料,揭示健康人群ANA分布规律,进一步探讨健康人群筛查ANA重要意义。方法采用间接免疫荧光法( IIF)检测ANA,免疫印迹法检测15项特异性抗体。结果体检人群中,ANA滴度>1∶100阳性率为14.01%,其中男性阳性率为9.04%,女性阳性率为19.05%;ANA滴度>1∶320阳性率为5.93%,其中男性阳

  15. Prevalence of symptoms of systemic lupus erythematosus (SLE) and of fluorescent antinuclear antibodies associated with chronic exposure to trichloroethylene and other chemicals in well water

    Energy Technology Data Exchange (ETDEWEB)

    Kilburn, K.H.; Warshaw, R.H. (Univ. of Southern California, Los Angeles (United States))


    Criteria for the recognition of systemic lupus erythematosus (SLE) were applied to 362 subjects exposed to trichloroethylene, trichloroethane, inorganic chromium, and other chemicals in water obtained from wells in an industrially contaminated aquifer in Tucson, Arizona. Their antinuclear autoantibodies were measured by fluorescence (FANA) in serum. Ten patients with clinical SLE and/or other collagen-vascular diseases were considered separately. Results were compared to an Arizona control group, to published series, and to laboratory controls. Frequencies of each of 10 ARA symptoms were higher in exposed subjects than in any comparison group except those with clinical SLE. The number of subjects with 4 or more symptoms was 2.3 times higher compared to referent women and men. FANA titers > 1:80 was approximately 2.3 times higher in women but equally frequent in men as in laboratory controls. ARA score and FANA rank were correlated with a coefficient (cc) of .1251, r{sup 2} = .0205 in women and this correlation was almost statistically significant in men cc = .1282, r{sup 2} = .0253. In control men and women neither correlation was significant. Long-term low-dose exposure to TCE and other chemicals in contaminated well water significantly increased symptoms of lupus erthematosus as perceived by the ARA score and the increased FANA titers.

  16. Antibody Phage Library Screening Efficiency Measured by KD Values

    Institute of Scientific and Technical Information of China (English)

    WANG Hui-tang; SHAN Ya-ming; TANG Li-li; GAO Li-zeng; WANG Li-ping; LI Wei; LI Yu-xin


    An antibody phage library was screened with two target molecules, IFNα-2a and FGFR-GST, and the KD value of each round of panning was measured. It was found that the apparent KD values decreased along with each additional panning round, which indicates the increase of the binding affinity between the phage and the target molecules.This result shows that the KD value is a reliable intrinsic parameter and a new method for screening efficiency detection is thus provided.

  17. Explore the relationship between ANA spectrum, anti-cardiolipin antibodies, antinuclear antibody, anti-ds-DNA levels and activity of SLE%探讨ANA谱、抗心磷脂抗体、抗核抗体、抗ds-DNA水平与SLE活动性的关系

    Institute of Scientific and Technical Information of China (English)



    Objective:To investigate ANA spectrum anti-cardiolipin antibodies, antinuclear antibody anti-ds-DNA levels and the relationship between the activity of SLE.Methods:from January 2013 to June 2014 in our hospital Rheumatology SLE patients hospitalized for a total of 125 cases of SLE group, and select the corresponding period in our hospital for examination of a total of 30 cases of healthy people healthy group. Observed active SLE, SLE and healthy inactive group of anti-cardiolipin antibodies, antinuclear antibody, anti-ds-DNA positive rate case, SLE active, SLE and healthy inactive group anticardiolipin antibody concentrations.Results: Health group and inactive SLE anti-cardiolipin antibodies, antinuclear antibodies, anti-ds-DNA positive rate, the difference was not statistically significant (P> 0.05). ACA- IgG SLE activity period (19.21 ± 0.79) RU / ml, ACA- IgM (17.01 ± 0.76) RU / ml were significantly higher than the healthy group and inactive SLE, the difference was statistically significant (P 0.05). Anti-U1-nRNP antibody of active SLE, anti-Sm antibodies.Conclusions:ANA spectrum, anti-cardiolipin antibodies, antinuclear antibodies, anti-ds-DNA levels measured for patients with active SLE can effectively detect the disease, and thus a better prediction of relapse, and help guide treatment.%目的:研究探讨ANA谱﹑抗心磷脂抗体﹑抗核抗体﹑抗ds-DNA水平与SLE活动性的关系。方法:选择2013年1月至2014年6月我院风湿免疫科住院的SLE患者共125例为SLE组,同时选择同期在我院进行体检的健康者共30例为健康组。观察SLE活动期、SLE非活动期与健康组抗心磷脂抗体、抗核抗体、抗ds-DNA阳性率情况,SLE活动期、SLE非活动期与健康组抗心磷脂抗体浓度情况。结果: SLE活动期的ACA- IgG(19.21±0.79) RU/ml、ACA- IgM(17.01±0.76)RU/ml水平均显著高于健康组和SLE非活动期,差异有统计学意义(P0.05)。结论:ANA谱、抗心磷脂抗

  18. Analysis of the Results of Antinuclear Antibody Spectrum for Patients of Moyamoya%402例烟雾病抗核抗体谱检测结果分析

    Institute of Scientific and Technical Information of China (English)

    田曙光; 曾利军; 陈建魁; 于农; 宋世平; 尹秀云; 黄媛; 金欣; 左向华; 杜宇


    Objective: Look for the occurrence of moyamoya disease with autoimmune of certain links. Methods: 402 cases of moyamoya disease spectrum antinuclear antibod respectively. Results: 112 cases of anti-nuclear antibody spectrum result was positive disease, the positive rate was 27.86%, and the percentage of anti-dsDNA, anti-CB, ant B, anti Jo-1, anti-RO-52, anti-ScL-70, anti-RIB, anti-PM-sd, anti-nRNP, anti-PCr anti-M2 antibodies were 16.07%, 1.79%, 5.36%, 17.86%, 7.14%, 5.36%, 13.39%, 5.3< 11.61%, 6.25% and 8.93%, 8.04%, respectively. Conclusion: Autoimmune disease nu moyamoya disease.%目的:探讨烟雾病的发生与自身免疫性疾病是否存在一定的联系.方法:对402例烟雾病抗核抗体谱检测结果进行回顾性分析.结果:402例烟雾病确诊病例中,有112位患者抗核抗体谱检测结果中存在阳性结果,阳性率为27.86%.其中,抗dsDNA、抗CB、抗Sm、抗SS-A、抗SS-B、抗Jo-1、抗RO-52、抗ScL-70、抗RIB、抗PM -sd、抗nRNP、抗PCNA、抗Nuc、抗HI、抗M2抗体在阳性病例中所占百分比分别为16.07%、1.79%、5.36%、17.86%、7.14%、5.36%、13.39%、5.36%、7.14%、4.46%、2.68%、11.61%、6.25%、8.93%和8.04%.结论:烟雾病患者中抗核抗体阳性率较高,自身免疫性疾病可能是烟雾病发生的重要诱因之一.

  19. Original Approach for Automated Quantification of Antinuclear Autoantibodies by Indirect Immunofluorescence

    Directory of Open Access Journals (Sweden)

    Daniel Bertin


    Full Text Available Introduction. Indirect immunofluorescence (IIF is the gold standard method for the detection of antinuclear antibodies (ANA which are essential markers for the diagnosis of systemic autoimmune rheumatic diseases. For the discrimination of positive and negative samples, we propose here an original approach named Immunofluorescence for Computed Antinuclear antibody Rational Evaluation (ICARE based on the calculation of a fluorescence index (FI. Methods. We made comparison between FI and visual evaluations on 237 consecutive samples and on a cohort of 25 patients with SLE. Results. We obtained very good technical performance of FI (95% sensitivity, 98% specificity, and a kappa of 0.92, even in a subgroup of weakly positive samples. A significant correlation between quantification of FI and IIF ANA titers was found (Spearman's ρ=0.80, P<0.0001. Clinical performance of ICARE was validated on a cohort of patients with SLE corroborating the fact that FI could represent an attractive alternative for the evaluation of antibody titer. Conclusion. Our results represent a major step for automated quantification of IIF ANA, opening attractive perspectives such as rapid sample screening and laboratory standardization.

  20. 健康体检人群中抗核抗体的年龄及性别分布状况研究%Effect of age and gender on distribution of antinuclear antibodies in healthy population

    Institute of Scientific and Technical Information of China (English)

    钱培新; 王蕾


    目的:探讨健康人群的年龄与性别对抗核抗体(ANA)表达的影响。方法选取2013年6月至2014年6月在我院行常规体检的健康人群1167例,受试者知情下空腹采血间接免疫荧光法检测ANA表达,以滴度≥1:100为阳性。分析ANA在健康人群中的年龄及性别分布状况。年龄及性别与ANA阳性率的相关性采用Spearman分析。结果1167例健康受试者中ANA阳性者157例,阳性率为13.45%。随着年龄增长,ANA阳性率在健康人群中有明显增大的趋势;而在同一年龄区间,女性的阳性率明显大于男性(P<0.05)。Spearman分析显示,年龄与健康人群ANA阳性率呈正相关性(rs=0.310,P<0.05);性别与健康人群ANA阳性率呈正相关性(rs=0.608,P<0.01)。结论抗核抗体在健康人群中表达会受年龄和性别因素影响。%Objective To explore the effect of age and gender on distribution of antinuclear antibodies (ANA) in healthy population. Methods A total of 1 167 healthy people who had routine check-up between June 2013 and June 2014 in our hospital were selected. The informed participants were taken fasting blood samples to deter-mine ANA expression by indirect immunofluorescence assay (IFA). Titer≥1:100 was set as positive result. The effect of age and gender on distribution of antinuclear antibodies (ANA) in the healthy population were analyzed. The corre-lations between ANA positive and age or gender were analyzed by Spearman method. Results Among 1 167 partici-pants, 157 showed ANA positive and the positive rate of ANA was 13.45%. There was a significantly increased trend of ANA positive rate with age within the healthy population. The positive rate of female participants was apparently higher than that of male participants in the same age range (P<0.05). Spearman analysis showed that there was a posi-tive correlation between ANA positive rate and age (rs=0.310, P=0.037), and also between ANA positive rate and gen-der (rs=0

  1. Thyroid Peroxidase Antibody and Screening for Postpartum Thyroid Dysfunction

    Directory of Open Access Journals (Sweden)

    Mohamed A. Adlan


    Full Text Available Postpartum thyroid dysfunction (PPTD is a common disorder which causes considerable morbidity in affected women. The availability of effective treatment for hypothyroid PPTD, the occurrence of the disease in subsequent pregnancies and the need to identify subjects who develop long term hypothyroidism, has prompted discussion about screening for this disorder. There is currently no consensus about screening as investigations hitherto have been variable in their design, definitions and assay frequency and methodology. There is also a lack of consensus about a suitable screening tool although thyroid peroxidase antibody (TPOAb is a leading contender. We present data about the use of TPOAb in early pregnancy and its value as a screening tool. Although its positive predictive value is moderate, its sensitivity and specificity when used in early pregnancy are comparable or better compared to other times during pregnancy and the postpartum period. Recent studies have also confirmed this strategy to be cost effective and to compare favourably with other screening strategies. We also explore the advantages of universal screening.

  2. 线性免疫印迹法在抗核抗体间接免疫荧光法筛查阴性标本中的临床意义%Detection of specific anti-nuclear antibodies by line immunoassay in indirect immunofluorescence-negative serum samples

    Institute of Scientific and Technical Information of China (English)

    李俊; 胡朝军; 沈波; 徐巍; 吴春龙


    Objective To evaluate the application of line immunoassay for detection of specific anti-nuclear antibody (ANA) in indirect immnofluorescence (HF)- negative serum samples. Methods Total 461 HF- ANA- consecutive samples were tested by DF for 15 specific anti- nuclear antibodies and their clinical significance was analyzed. Results In 216 patients of autoimmune disease (AID) LIA- ANAs+ was identified in 100 serum samples with a positive rate of 46.30%, which was higher than that of non- AID patients (20.00%). In AID patients the positive rate of LIA(3+), LIA(2+), LIA(1 + ) and LIA(± ) were higher than those of non- AID patients (P0.05). The positive rate of anti- Sm and anti- dsDNA in systemic lupus erythematosus (SLE) were higher than those of non- SLE (P<0.05). Conclusion It is recommend to detect serum specific antinuclear antibodies with line immunoassay whether the ANA is positive or negative shown by screening assay of DF.%目的 探讨采用线性免疫印迹法(LIA)在抗核抗体(ANA)间接免疫荧光法(IIF)筛查阴性标本中进行抗核抗体谱(ANAs)特异性抗体检测的价值和临床意义.方法 对461例IIF-ANA阴性临床血清标本采用线性免疫印迹法进行15种ANAs特异性抗体(LIA-ANAs)检测.结果 216例自身免疫性疾病(AID)组标本中LIA-ANAs阳性数为100例,占46.30%,高于非AID组(20.00%);AID组LIA-ANAs阳性结果按灰度强度分,LIA(3+)、LIA(2+)、LIA(1+) 、LIA(±)的阳性率均高于非AID组,差异均有统计学意义(均P<0.05).AID组中ANAs特异性抗体中的抗Ro-52、抗SS-A、抗Sm、抗nRNP/Sm、抗SS-B抗体、抗dsDNA和抗rRNP抗体的阳性率分别为2.31%~18.06%,高于非AID组,差异有统计学意义(P<0.05).AID组中ANAs特异性抗体阳性强度LIA(1+)+LIA(±)的阳性率高于LIA(3+)+LIA(2+)的阳性率,差异有统计学意义(P<0.01).抗nRNP/Sm、抗Ro-52、抗SS-A和抗SSB抗体在AID组中阳性率的差异无统计学意义(P >0.05);系统性红斑狼疮(SLE)与非SLE比

  3. [Comparison of eight screening tests for ant-HCV antibody]. (United States)

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


    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".

  4. Analysis of the results of antinuclear antibody and anti-ENAantibodies for Han people in Shihezi area xinjiang province%新疆石河子地区汉族人群ANA与抗ENA 抗体的检测结果分析

    Institute of Scientific and Technical Information of China (English)

    何建伟; 李静; 赵涛; 曹薇薇


    Objective To analyze the relationship between antinuclear antibody (ANA) and anti-extractable nuclear antigens (ENA) antibodies fron H an people in Shihezi area Xin jiang province .Methods ANA and anti-ENA antibodies were detected by indirect immunofluorescence (IIF) and EUROLINE-Western B lot technology respectively .Double blind m ethod was perform ed to analyze the relationship .Results 0f all the 1084 samples,399 cases were positive for ANA ,the positive tate was 36.81% (399/1084) .512 cases in the above collected sam ples were tested by IIF and EUR0LINE-W estern B lot technology sim ultaneously .The majority of patterns were nuclear hom ogeneous patrern (122 cases) ,nuclear speckled patterns(31 cases) ,nuclear peripheral patterns(12 cases),mixed patterns(4 cases) .Of the 161 positive anti-ENA antibodies cases,eighty were negative ANA cases.Of the 351 negative antiENA antibodies cases,eighty-eightwere positive ANA cases. In the cases of nuclear hom ogeneous pattern for ANA ,the commonantiENA antibodies were anti-Ro/SSA 60 (35 cases).In the cases of nuclear speckled patterns for ANA ,the commonanti-ENA antibodies were anti-Ro/SSA60 (14 cases),anti-UlRNP (7 cases),anti-Sm (9 cases).In the cases of nuclear peripheral patterns for ANA ,the common anti-ENA antibodies were anti-Ro/SSA 60 (5 cases) ,anti-dsDNA (6 cases), anti-AnuA (5 cases)、 anti-histon (5 cases).Conclusion The Coincidence level between ANA and anti-ENA antibodies results was not fine.There was not any fixed regularity between ANA and anti-ENA antibodies results.In order to accurately determ ine the tager antigens,it is necesary to screen ANA with IF and to confirm the types by anti-ENA antibodies.M any new anti-ENA antibodieswere expected to be puried in the future.%目的 回顾本实验室开展抗核抗体(antinuclear antibody,ANA)与抗可提取性核抗原(extractable nuclear antigens,ENA)抗体联合检测的情况,分析新疆石河子地区汉族人群中ANA与抗ENA

  5. Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library. (United States)

    Wang, Han; Yu, Rui; Fang, Ting; Yu, Ting; Chi, Xiangyang; Zhang, Xiaopeng; Liu, Shuling; Fu, Ling; Yu, Changming; Chen, Wei


    Tetanus neurotoxin (TeNT) produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc) as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H) can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro. Moreover, 2-7G inhibited TeNT-Hc binding to the receptor via carbohydrate-binding sites of the W pocket while 2-2D and S-4-7H inhibited binding of the R pocket. Although no single mAb completely protected mice from the toxin, they could both prolong survival when challenged with 20 LD50s (50% of the lethal dose) of TeNT. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, indicating their high neutralizing potency in vivo. Antibodies recognizing different carbohydrate-binding pockets could have higher synergistic toxin neutralization activities than those that recognize the same pockets. These results could lead to further production of neutralizing antibody drugs against TeNT and indicate that using TeNT-Hc as an antigen for screening human antibodies for TeNT intoxication therapy from human immune antibody library was convenient and effective.

  6. Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library (United States)

    Wang, Han; Yu, Rui; Fang, Ting; Yu, Ting; Chi, Xiangyang; Zhang, Xiaopeng; Liu, Shuling; Fu, Ling; Yu, Changming; Chen, Wei


    Tetanus neurotoxin (TeNT) produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc) as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H) can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro. Moreover, 2-7G inhibited TeNT-Hc binding to the receptor via carbohydrate-binding sites of the W pocket while 2-2D and S-4-7H inhibited binding of the R pocket. Although no single mAb completely protected mice from the toxin, they could both prolong survival when challenged with 20 LD50s (50% of the lethal dose) of TeNT. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, indicating their high neutralizing potency in vivo. Antibodies recognizing different carbohydrate-binding pockets could have higher synergistic toxin neutralization activities than those that recognize the same pockets. These results could lead to further production of neutralizing antibody drugs against TeNT and indicate that using TeNT-Hc as an antigen for screening human antibodies for TeNT intoxication therapy from human immune antibody library was convenient and effective. PMID:27626445

  7. Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library

    Directory of Open Access Journals (Sweden)

    Han Wang


    Full Text Available Tetanus neurotoxin (TeNT produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro. Moreover, 2-7G inhibited TeNT-Hc binding to the receptor via carbohydrate-binding sites of the W pocket while 2-2D and S-4-7H inhibited binding of the R pocket. Although no single mAb completely protected mice from the toxin, they could both prolong survival when challenged with 20 LD50s (50% of the lethal dose of TeNT. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, indicating their high neutralizing potency in vivo. Antibodies recognizing different carbohydrate-binding pockets could have higher synergistic toxin neutralization activities than those that recognize the same pockets. These results could lead to further production of neutralizing antibody drugs against TeNT and indicate that using TeNT-Hc as an antigen for screening human antibodies for TeNT intoxication therapy from human immune antibody library was convenient and effective.

  8. Results analysis of antinuclear antibodies spectrum tests in four kinds of autoimmune diseases%自身免疫性疾病的抗核抗体谱检测

    Institute of Scientific and Technical Information of China (English)

    张园园; 潘宝龙; 马骏; 李俊娥


    目的:探讨抗核抗体谱(ANAs)检测在系统性红斑狼疮(SLE)、干燥综合征(SS)、混合性结缔组织病(MCTD)、类风湿性关节炎(RA)患者中的诊断价值。方法回顾性分析1359例自身免疫性疾病患者血清抗核抗体谱16项检测,对单个抗体在4种疾病患者中的阳性率每种疾病中ANAs的构成比进行分析。结果 ANA在前3种疾病中阳性率均高于RA;抗ds-DNA、抗nRNP主要见于SLE和MCTD;抗Sm主要见于SLE;抗SSA、抗Ro-52、抗SSB阳性率表现为SS>MCTD>SLE>RA;抗SCL-70、抗PM-SCL、抗Jo-1均为较低阳性率;其余自身抗体阳性率表现为SLE>MCTD>SS≈RA。在数据基础上形成了4种自身免疫性疾病诊断的ANAs阳性率参考模型。结论可根据ANA谱中单个抗体在4种疾病中的阳性率特点,建立4种自身免疫性疾病诊断和鉴别诊断的ANAs阳性率参考模型。%ObjectiveTo explore the value of antinuclear antibodies(ANAs) spectrum tests in SLE、SS、MCTD and RA.Methods Retrospectively analyzed ANAs test results of 1359 cases patients with autoimmune diseases in our hospital since January 2011, made statistical analysis on positive rate of each autoantibody in four kinds of disease and ANAs constituent ratio in each disease.Results The positive rate of each autoantibody had certain characteristics :(1)The ANA positive rate in the previous three diseases reached more than 92.8%, while somewhat less in RA (48.5%);(2)Anti-dsDNA and anti-nRNP were mainly in SLE and MCTD;(3)Anti-Sm was mainly in SLE(26.5%);(4)The positive rates of anti-SSA,anti-Ro-52 and anti-SSB showed: SS>MCTD>SLE>RA; (5)The positive rates of anti-SCL70, anti-PM-SCL and anti-Jo1 were lower, no significant difference;(6)The other remaining autoantibodies were: SLE>MCTD>SS≈RA. The diagnostic reference model of ANAs constituent ratio of the four kinds of autoimmune disease: (1)SLE: ANA (93.1%), Ro-52 (50.4%), SSA (47.3%), Ans (42.5%), anti-dsDNA (41

  9. Evaluation of screening tests for the detection of antistreptolysin O antibodies.


    Lue, Y. A.; Nicolas, C.; Kemawikasit, P.; Pierre, A.; McLean, T I; Simms, D H


    The accuracy of two screening tests, one utilizing serum and the other utilizing whole blood, was compared with the accuracy of the conventional macrotitration method for the detection of antistreptolysin O antibodies. Of the 569 specimens tested with the serum screening procedure and the macrotitration method, 235 and 282, respectively, were positive for antistreptolysin O antibodies. Comparative testing of 200 specimens with the peripheral blood screening test and the macrotitration method ...

  10. Yeast surface display for antibody isolation: library construction, library screening, and affinity maturation. (United States)

    Van Deventer, James A; Wittrup, Karl Dane


    Antibodies play key roles as reagents, diagnostics, and therapeutics in numerous biological and biomedical research settings. Although many antibodies are commercially available, oftentimes, specific applications require the development of antibodies with customized properties. Yeast surface display is a robust, versatile, and quantitative method for generating these antibodies and is accessible to single-investigator laboratories. This protocol details the key aspects of yeast surface display library construction and screening.

  11. A Strategy for Screening Monoclonal Antibodies for Arabidopsis Flowers (United States)

    Shi, Qian; Zhou, Lian; Wang, Yingxiang; Ma, Hong


    The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants. PMID:28293248

  12. NanoLuc luciferase - A multifunctional tool for high throughput antibody screening

    Directory of Open Access Journals (Sweden)

    Nicolas eBoute


    Full Text Available Based on the recent development of NanoLuc Luciferase a small (19 kDa, highly stable, ATP independent, bioluminescent protein, an extremely robust and ultra high sensitivity screening system has been developed whereby primary hits of therapeutic antibodies and antibody fragments could be characterized and quantified without purification. This system is very versatile allowing cellular and solid phase ELISA but also homogeneous BRET based screening assays, relative affinity determinations with competition ELISA and direct western blotting. The new NanoLuc Luciferase protein fusion represents a swiss army knife solution for today and future high throughput antibody drug screenings.

  13. Clustering analysis of anti-nuclear antibodies in systemic lupus erythematosus%系统性红斑狼疮患者抗核抗体谱聚类分析初探

    Institute of Scientific and Technical Information of China (English)

    陆晓东; 单小云; 赵硕; 杜红卫; 薛亚东


    Objective To examine clusters of anti-nuclear antibodies (ANA) and their associations with clinical features in patients with systemic lupus erythematosus (SLE).Methods It was a retrospective study.113 SLE patients were reviewed from March 2010 to May 2012 in Department of Rheumatology,Jinhua Central Hospital.ANA and specific autoantibodies to 15 kinds of nuclear antigens were tested by indirect immunofluorescence assay (IIF) and line immunoassay (LIA) respectively.Hierarchical clustering method was performed to analyze specific clusters of ANA profiles in SLE.Chi-square tests were used to investigate relationship between antibody clusters and clinical features of SLE.Results The positive rate of LIA for ANA was 97.3%,consistent with IIF method,and the total accordance rate of the both methods was 98.2%.Thirteen kinds of antigen-specific antibodies were detected in SLE patients by LIA.Clustering analysis for these antibodies showed three specific clusters in SLE,Nuc/His/dsDNA cluster (C1),low-Ro/low-La cluster (C2),and Ro/Sm/RNP cluster (C3),accounting for 36.3%,24.8%,38.9% of the total cases respectively.There were significant difference of AST levels among three clusters [(32.62 ± 21.92)U/L,(25.56 ± 16.63) U/L,(50.41 ± 60.86) U/L respectively for C1,C2 and C3].High incidences of chronic cutaneous lupus,abnormal renal indicators and inflammatory synovitis were found in all three clusters.Besides,there were significant differences among three clusters for the incidences of chronic cutaneous lupus (39.0%,39.3%,63.6% respectively for C1,C2,C3) and leukopenia/lymphopenia (56.1%,25.0%,56.8% respectively for C1,C2,C3) (P < 0.05).Patients in Ro/Sm/RNP cluster showed higher incidences of lupus nephritis (43.2%/26.8% or 39.3%); patients in low-Ro/low-La cluster showed low risk of hypertension (7.1%/19.5% or 22.7%) ; patients in Nuc/His/dsDNA cluster showed high incidences of thrombocytopenia (41.5%/21.4% or 25.0%) and high risk

  14. 抗核抗体荧光模型特异性抗体谱及滴度与自身免疫性疾病的关联性研究%Study on the relationship between antinuclear antibody fluorescence model, specific antibodies spectrum, ti-ter and autoimmune diseases

    Institute of Scientific and Technical Information of China (English)

    李炎梅; 莫思健; 梁太英; 莫显文; 覃平; 何玮; 莫飞; 朱金月


    Objective To investigate the relationship between antinuclear antibody ( ANA ) fluorescence model, specific antibodies spectrum, titer and autoimmune diseases(AID).Methods Four hundred and four pa-tients with AID were investigated by using indirect immunofluorescence assay to detect ANA, at the same time, using Europe and Mongolia blotting to detect 15 specific antinuclear antibodies spectrum ( ANAS), and a comparative analy-sis was performed of expression of ANA fluorescence model and the titer and ANA fluorescence model in different dis -eases.Results In AID group the positive rate of ANA was 68.81%, titers in 1∶1 000 accounted for 75.90%.ANA fluorescence model mainly showed cytoplasmic type and nuclear homogeneous type, 130 cases and 108 cases respec-tively.In ANA positive patients, the titer of ANA was higher and fluorescent hybrid model was also increased. Con-clusion The detection rate of ANA was higher in patients with AID. In fluorescent ANA positive patients with high titer the fluorescence model mainly is hybrid model. The simultaneous detection of ANA and ANAS has important clinical significance for diagnosis of autoimmune diseases.%目的:探讨抗核抗体( ANA)荧光模型、滴度及特异性抗体谱与自身免疫性疾病的关联性。方法对404例自身免疫性疾病( AID)患者,同时采用间接免疫荧光法检测ANA和欧蒙印迹法检测15项特异性抗核抗体谱( ANAS),对比分析ANA荧光模型和滴度以及ANA荧光模型在不同疾病中的表达,并与健康体检者300名进行比较。结果在AID组中的ANA检出阳性率为68.81%,滴度在1∶1000以上者占75.90%。检出的ANA荧光模型,以胞浆型和核均质型为主,分别检出130例和108例。 ANA阳性患者滴度升高,检出荧光混合模型的比例也明显增加。结论 AID患者中ANA检出率较高,高滴度的ANA阳性患者荧光模型以混合模型为主,同时检测ANA和ANAS对自身免疫性疾病诊断具有重要的临床意义。

  15. Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris. (United States)

    Shah, Kartik A; Clark, John J; Goods, Brittany A; Politano, Timothy J; Mozdzierz, Nicholas J; Zimnisky, Ross M; Leeson, Rachel L; Love, J Christopher; Love, Kerry R


    Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria, or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, "hit-to-lead identification" remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigen-specific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV).

  16. The test results analysis of antinuclear antibodies using different brands kit and methodology%不同品牌试剂盒及方法学间抗核抗体检测结果分析

    Institute of Scientific and Technical Information of China (English)

    赵迪; 陈燚琼; 邓晓刚; 杨文杰; 徐雨杰; 李智


    Objective To compare test results of antinuclear antibodies (ANA ) using different brands kit and methodology . Methods 180 cases who accepted serum ANA detection in hospital from April to December 2012 were choosen ,in which 145 cases of positive serum and 35 cases of healthy control .The 180 cases were tested for ANAs by YHLO ANA-17S ,EUROIMMUN ANA profile 3 and ANA IIFA kits at the same time ,and all parameters about ANA decetion were compared such as sensitivity ,specifici-ty ,positive predictive value ,negative predictive value and the coincidence rate between the two kinds of kits .Results The sensitivi-ty of YHLO ANA-17S ,EUROIMMUN ANA profile 3 and EUROIMMUN ANA IIFA was 94 .8% ,90 .4% ,95 .6% ;specificity was 88 .9% ,91 .1% ,93 .3% ;positive predictive value was 96 .2% ,96 .8% ,97 .7% ;negative predictive value was 85 .1% ,75 .9% , 87 .5% ;the coincidence rate between the EUROIMMUN ANA profile 3 and EUROIMMUN ANA IIFA was 84 .0% (P>0 .05) ,be-tween the YHLO ANA-17S and EUROIMMUN ANA IIFA was 80 .0% (P>0 .05) ,between theYHLO ANA-17S and EUROIM-MUN ANA profile 3 was 93 .0% (P> 0 .05) .Conclusion Three kits for detecting ANAs have good sensitivity and specificity , which can be used for autoimmune disease detection .Laboratory tests result should be combined with clinical symptoms according to the diagnostic criteria to make the final diagnosis .%目的:比较分析不同品牌试剂盒及方法学间抗核抗体检测结果。方法选取该院2012年4~12月进行自身抗体检测血清者180例,其中自身免疫性疾病患者145例,健康对照35例;同时进行亚辉龙ANA-17S、欧蒙ANA谱3以及欧蒙IIFA的平行比对试验。分别计算灵敏度、特异度、阳性预测值、阴性预测值及两种试剂盒间的符合率。结果亚辉龙ANA-17S、欧蒙ANA谱3、欧蒙IIFA的灵敏度依次为94.8%、90.4%、95.6%;特异度依次为88.9%、91.1%、93.3%;阳性预测值依次为96.2%、96.8%、97

  17. High-efficiency screening of monoclonal antibodies for membrane protein crystallography.

    Directory of Open Access Journals (Sweden)

    Hyun-Ho Lim

    Full Text Available Determination of crystal structures of membrane proteins is often limited by difficulties obtaining crystals diffracting to high resolution. Co-crystallization with Fab fragments of monoclonal antibodies has been reported to improve diffraction of membrane proteins crystals. However, it is not simple to generate useful monoclonal antibodies for membrane protein crystallography. In this report, we present an optimized process for efficient screening from immunization to final validation of monoclonal antibody for membrane protein crystallography.

  18. Women's attitude towards prenatal screening for red blood cell antibodies, other than RhD

    Directory of Open Access Journals (Sweden)

    van der Schoot CE


    Full Text Available Abstract Background Since July 1998 all Dutch women (± 200,000/y are screened for red cell antibodies, other than anti-RhesusD (RhD in the first trimester of pregnancy, to facilitate timely treatment of pregnancies at risk for hemolytic disease of the fetus and newborn (HDFN. Evidence for benefits, consequences and costs of screening for non-RhD antibodies is still under discussion. The screening program was evaluated in a nation-wide study. As a part of this evaluation study we investigated, according to the sixth criterium of Wilson and Jüngner, the acceptance by pregnant women of the screening program for non-RhD antibodies. Methods Controlled longitudinal survey, including a prenatal and a postnatal measurement by structured questionnaires. Main outcome measures: information satisfaction, anxiety during the screening process (a.o. STAI state inventory and specific questionnaire modules, overall attitude on the screening program. Univariate analysis was followed by standard multivariate analysis to identify significant predictors of the outcome measures. Participants: 233 pregnant women, distributed over five groups, according to the screening result. Results Satisfaction about the provided information was moderate in all groups. All screen- positive groups desired more supportive information. Anxiety increased in screen- positives during the screening process, but decreased to basic levels postnatally. All groups showed a strongly positive balance between perceived utility and burden of the screening program, independent on test results or background characteristics. Conclusion Women highly accept the non-RhD antibody screening program. However, satisfaction about provided information is moderate. Oral and written information should be provided by obstetric care workers themselves, especially to screen-positive women.

  19. Protein crystallization with microseed matrix screening: application to human germline antibody Fabs

    Energy Technology Data Exchange (ETDEWEB)

    Obmolova, Galina, E-mail:; Malia, Thomas J.; Teplyakov, Alexey; Sweet, Raymond W.; Gilliland, Gary L., E-mail: [Janssen Research and Development LLC, 1400 McKean Road, Spring House, PA 19477 (United States)


    The power of microseed matrix screening is demonstrated in the crystallization of a panel of antibody Fab fragments. The crystallization of 16 human antibody Fab fragments constructed from all pairs of four different heavy chains and four different light chains was enabled by employing microseed matrix screening (MMS). In initial screening, diffraction-quality crystals were obtained for only three Fabs, while many Fabs produced hits that required optimization. Application of MMS, using the initial screens and/or refinement screens, resulted in diffraction-quality crystals of these Fabs. Five Fabs that failed to give hits in the initial screen were crystallized by cross-seeding MMS followed by MMS optimization. The crystallization protocols and strategies that resulted in structure determination of all 16 Fabs are presented. These results illustrate the power of MMS and provide a basis for developing future strategies for macromolecular crystallization.

  20. Retrospective analysis of antinuclear antibody test results of 4188 specimens%临床标本抗核抗体4188份检测结果回顾性分析

    Institute of Scientific and Technical Information of China (English)

    杨银忠; 陈敏敏; 程文霞; 龙海燕


    目的:探讨抗核抗体(ANA)常见类型在四川地区自身免疫性疾病(AID)患者中的分布趋势及临床价值。方法采用免疫印迹法对2011年1月至2013年9月4188份疑似AID患者的临床标本进行ANA常见15种类型(抗NRNP、抗SM、抗SS-A、抗RO-52、抗SS-B、抗SCL-70、抗PM-SCL、抗JO-1、抗CENP、抗PCNA、抗ds-DNA、抗组蛋白、抗核小体、抗核糖体P蛋白及抗AMAM2)的检测,统计其分布情况,并对性别、年龄及季节进行分组,采用χ2检验进行统计学分析。结果(1)在4188份标本中,ANA阳性率为28.8%,AID阳性率为7.2%。在确诊的303例AID患者中,干燥综合征(SS)患者抗SS-A阳性率(75.4%)最高,依次为抗RO-52(61.5%)及抗SS-B(20.5%);系统性红斑狼疮(SLE)患者抗SS-A阳性率(76.8%)最高,依次为抗 RO-52(64.2%)及抗 ds-DNA (41.1%);混合性结缔组织(MCTD)患者抗 RO-52阳性率(42.5%)最高,其次为抗SS-A(41.1%)及抗NRNP(26.0%);类风湿性关节炎(RA)患者抗SM阳性率(65.4%)最高,其次为抗SS-A(38.5%)。SS是四川地区最常见的AID,依次为SLE、MCTD及RA等。(2)在4188例疑似AID患者中,女性AID阳性率(11.0%)显著高于男性(1.5%),差异有统计学意义(χ2=136.44,P<0.05);年龄组中18~45岁组阳性率最高(8.4%),<18岁组最低(2.1%),差异有统计学意义(χ2=8.92,P<0.05);季度组中第4季度阳性率最高(8.6%),第1季度最低(6.2%),差异有统计学意义(χ2=3.47,P<0.05)。结论四川地区AID患者ANA常见类型分布可能与年龄、季节及性别相关,这在AID的诊断方面有重要价值。%Objective To explore the distribution trend and clinical value of antinuclear antibody (ANA) in patients with autoimmune diseases ( AID ) in Sichuan region . Methods Immunoblotting test was employed among 4 188 specimens with suspected AID from January

  1. Characteristic analysis of the clinical expressions of antinuclear antibody and antibody profiles in com-mon joint diseases%抗核抗体及抗体谱在常见关节疾病中的临床表达特征分析

    Institute of Scientific and Technical Information of China (English)

    王家驷; 孙良丽; 焦鑫; 吴建红; 王廷杰; 何思春


    Objective To explore the clinical significance of antinuclear antibody (ANA) and antinu-clear antibody spectrum (ANAs) detecting in the diagnosis of common joint diseases. Methods The clinical data of 452 cases patients with joint disease in our hospital from January 2013 to October 2014 were collected and retrospectively analyzed. The patients were divided into osteoarthropathy group 63 cases , rheumatoid arthritis(RA) group 291 cases and gouty arthritis group 98 cases according to the clinical diagnosis. The distri-butions of ANA and ANAs in different disease groups were analyzed respectively. The different expression combinations of ANAs among different diseases, as well as the differential expressions of the ANA fluorescent types among different disease groups were further analyzed. The results were analyzed statistically. Results (1)There was 170 cases (37.6%,170/452) patients showed ANA+, 88 cases (19.5%,88/452) ANA+/ANAs+, 82 cases (18.1%,82/452) ANA+/ANAs-, and no ANA-/ANAs+was found among 452 cases pa-tients. There was statistical significance in the difference of ANA and ANAs results (χ2=177.98, P=0.000). (2)There were 18 cases(28.6%,18/63) patients with osteoarthropathy showed ANA+, and found 6 kinds flu-orescent types, of which cytoplasm was the most(44.4%,8/18);There were 146 cases(50.2%,146/291) pa-tients with RA showed ANA+, and found 13 kinds fluorescent types, of which the homogeneous type (32.2%, 47/146) was the most; There were 6 cases (6.1%,6/98) patients with gouty arthritis showed ANA+, and found 5 kinds fluorescent types, all of which appeared in no more than 2 cases;There was statistical signifi-cance in the ANA positive rate among three joint disease groups (P=0.000), and some fluorescent types of ANA were associated with different joint diseases(χ2=21.43, P=0.018). (3)There were 6 cases(9.5%,6/63) patients with osteoarthropathy showed ANAs+, and found 6 kinds antibodies. There were 78 cases (26.8%,78/291) patients with RA showed ANAs

  2. Screening a hybridoma producing a specific monoclonal antibody to HLA-A24+Bw4 antigen by cytotoxicity inhibition assay. (United States)

    Hiroishi, S; Kaneko, T; Arita, J


    A hybridoma secreting a monoclonal antibody (Tsa-1, IgG3) reacting specifically to HLA-A24+Bw4 was screened by cytotoxicity inhibition assay and micrototoxicity test. The R value of the antibody was 0.843.

  3. The Statistical Precision of Medical Screening Procedures: Application to Polygraph and AIDS Antibodies Test Data


    Gastwirth, Joseph L.


    The increased use of screening tests for drug use or antibodies to the HTLV-III (AIDS) virus, as well as pre-employment polygraph testing, has raised concerns about the reliability of the results of these procedures. This paper reviews the mathematical model underlying the analysis of data from screening tests. In addition to the known formulas for the proportion of positive (negative) classifications that are correct, we provide a large sample approximation to their standard errors. The resu...

  4. Surface plasmon resonance biosensing: Approaches for screening and characterising antibodies for food diagnostics. (United States)

    Yakes, B J; Buijs, J; Elliott, C T; Campbell, K


    Research in biosensing approaches as alternative techniques for food diagnostics for the detection of chemical contaminants and foodborne pathogens has increased over the last twenty years. The key component of such tests is the biorecognition element whereby polyclonal or monoclonal antibodies still dominate the market. Traditionally the screening of sera or cell culture media for the selection of polyclonal or monoclonal candidate antibodies respectively has been performed by enzyme immunoassays. For niche toxin compounds, enzyme immunoassays can be expensive and/or prohibitive methodologies for antibody production due to limitations in toxin supply for conjugate production. Automated, self-regenerating, chip-based biosensors proven in food diagnostics may be utilised as rapid screening tools for antibody candidate selection. This work describes the use of both single channel and multi-channel surface plasmon resonance (SPR) biosensors for the selection and characterisation of antibodies, and their evaluation in shellfish tissue as standard techniques for the detection of domoic acid, as a model toxin compound. The key advantages in the use of these biosensor techniques for screening hybridomas in monoclonal antibody production were the real time observation of molecular interaction and rapid turnaround time in analysis compared to enzyme immunoassays. The multichannel prototype instrument was superior with 96 analyses completed in 2h compared to 12h for the single channel and over 24h for the ELISA immunoassay. Antibodies of high sensitivity, IC50's ranging from 4.8 to 6.9ng/mL for monoclonal and 2.3-6.0ng/mL for polyclonal, for the detection of domoic acid in a 1min analysis time were selected. Although there is a progression for biosensor technology towards low cost, multiplexed portable diagnostics for the food industry, there remains a place for laboratory-based SPR instrumentation for antibody development for food diagnostics as shown herein.

  5. Early developability screen of therapeutic antibody candidates using Taylor dispersion analysis and UV area imaging detection. (United States)

    Lavoisier, Alexandra; Schlaeppi, Jean-Marc


    Therapeutic antibodies represent one of the fastest growing segments in the pharmaceutical market. They are used in a broad range of disease fields, such as autoimmune diseases, cancer, inflammation and infectious diseases. The growth of the segment has necessitated development of new analytical platforms for faster and better antibody selection and characterization. Early quality control and risk assessment of biophysical parameters help prevent failure in later stages of antibody development, and thus can reduce costs and save time. Critical parameters such as aggregation, conformational stability, colloidal stability and hydrophilicity, are measured during the early phase of antibody generation and guide the selection process of the best lead candidates in terms of technical developability. We report on the use of a novel instrument (ActiPix/Viscosizer) for measuring both the hydrodynamic radius and the absolute viscosity of antibodies based on Taylor dispersion analysis and UV area imaging. The looped microcapillary-based method combines low sample consumption, fast throughput and high precision compared to other conventional methods. From a random panel of 130 antibodies in the early selection process, we identified some with large hydrodynamic radius outside the normal distribution and others with non-Gaussian Taylor dispersion profiles. The antibodies with such abnormal properties were confirmed later in the selection process to show poor developability profiles. Moreover, combining these results with those of the viscosity measurements at high antibody concentrations allows screening, with limited amounts of materials, candidates with potential issues in pre-formulation development.

  6. An antibody microarray, in multiwell plate format, for multiplex screening of foodborne pathogenic bacteria and biomolecules (United States)

    Intoxication and infection caused by foodborne pathogens are important problems in the United States, and screening tests for multiple pathogen detection have been developed because food producers are known reservoirs of multiple pathogens. We developed a 96-well microplate, multiplex antibody micr...

  7. 血清抗核抗体在视神经脊髓炎谱系疾病和多发性硬化中的分布%Distribution of antinuclear antibodies in patients with neuromyelitis optica spectrum disorders and multiple sclerosis

    Institute of Scientific and Technical Information of China (English)

    武雷; 黄德晖; 杨扬; 吴卫平


    目的 研究血清抗核抗体(ANAs)在视神经脊髓炎谱系疾病(NMOSDs)和多发性硬化(MS)中的分布.方法 收集2009-01-2011-03间在作者医院神经内科门诊和住院诊治并行血清ANAs筛查的NMOSDs患者74例,包括视神经脊髓炎(NMO)53例、复发长节段横贯性脊髓炎(rLETM)20例和复发性视神经炎(RON)1例,以及MS患者49例,统计其血清ANAs阳性率并进行分析.结果 NMOSDs患者血清ANAs阳性率为45.9%(34/74),其中ANA(本文中特指用间接免疫荧光法检测的抗核抗体)、抗dsDNA、抗着丝粒抗体(ACA)、抗SSA抗体、抗SSB抗体阳性率分别为36.5%(27/74)、5.4%(4/74)、1.4%(1/74)、27.0%(20/74)、9.5%(7/74),MS组仅1例ANAs阳性,阳性率为2.0%(1/49),两组间差异有统计学意义(P<0.01).血清ANAs诊断NMOSDs的灵敏度为45.9%,特异度达98.0%;NMO和rLETM患者血清ANAs阳性率分别为47.2%和40.0%,两者无统计学差异(P=0.635).结论 NMO和rLETM患者血清ANAs阳性率高于MS组,支持NMO和rLETM同属于NMOSDs的观点.ANAs有可能是NMOSDs和MS两组疾病的鉴别指标之一.%Objective To investigate the seroprevalence of antinuclear antibodies (ANAs) in patients with neuromyelitis optica spectrum disorders (NMOSDs) and multiple sclerosis (MS). Methods Seventy-four patients with NMOSDs and 49 with MS were screened for serum ANAs and enrolled in the study. They were in hospital from January 2009 to March 2011. The NMOSDs group included 53 patients with neuromyelitis optica (NMO), 20 with recurrent longitudinally extensive transverse myelitis (rLETM), 1 with recurrent optic neuritis (RON) and 49 with MS. Seroprevalence of ANAs was analyzed. Results Seroprevalence of ANAs in NMOSDs patients was 45.9% (34/74), of which the seroprevalence rates of ANA, anti-dsDNA, anti-centromere antibody (ACA), anti-SSA and anti-SSB were 36.5% (27/74), 5.4% (4/74), 1.4% (1/74), 27.0% (20/74) and 9. 5% (7/74), respectively. In the MS group, only 1 patient showed seropositive

  8. The clinical significance comparison of a latex agglutination based syphilis screening test at low antibody titer. (United States)

    Wang, Hua-Cheng; Chen, Cha; Wang, Li-Na; Long, Yi-Fei; Zhang, Wei-Zheng; Li, You-Qiang; Xiao, Qian; Yuan, Hui


    The rapid increase of syphilis underscores a tremendous need to carefully evaluate many new serological tests for syphilis and choose efficient and economical strategies for syphilis screening, especially in the case of primary infection with low antibody titer. Between 2011 and 2012, 73 patients' sera samples were included in this retrospective study. They were either TRUST or TPPA reactive, either LA (latex agglutination) based auto3 TP or CLIA (chemiluminescence assay) based Architect Syphilis TP assay reactive. The contradictory weak response samples were further examined by FTA-Abs method. TPPA could not give reactive results in samples with antibody concentration less than 10 mIU. Auto3 TP reagent shows good linearity at low antibody titers and was more sensitive than TPPA, while the former does not show significant superiority compared to the Architect Syphilis TP assay at low antibody titer, except that it is suitable for adaptation on diverse automated chemistry analyzers.

  9. Antibodies against G-protein coupled receptors: novel uses in screening and drug development. (United States)

    Gupta, Achla; Heimann, Andrea S; Gomes, Ivone; Devi, Lakshmi A


    Antibodies are components of the body's humoral immune system that are generated in response to foreign pathogens. Modern biomedical research has employed these very specific and efficient molecules designed by nature in the diagnosis of diseases, localization of gene products as well as in the rapid screening of targets for drug discovery and testing. In addition, the introduction of antibodies with fluorescent or enzymatic tags has significantly contributed to advances in imaging and microarray technology, which are revolutionizing disease research and the search for effective therapeutics. More recently antibodies have been used in the isolation of dimeric G protein-coupled receptor (GPCR) complexes. In this review, we discuss antibodies as powerful research tools for studying GPCRs, and their potential to be developed as drugs themselves.

  10. Relationship and Nursing Countermeasures of Antinuclear Antibodies, Liver Specific Autoantibodies with Chronic Hepatitis B%抗核抗体、自身免疫性抗体与慢性乙肝的关系及护理对策分析

    Institute of Scientific and Technical Information of China (English)

    刘育琼; 魏桂菊; 何秋娴


    Objective:To study relationship and nursing countermeasures of antinuclear antibodies, liver specific autoantibodies with chronic hepatitis B.Method:From March 2013 to March 2014 in our hospital for treatment of 100 cases of patients with chronic hepatitis B (CHB) as the research object.Chose 92 cases in our hospital received physical examination of normal persons as control group, the related antibodies, CHB from positive and negative group of related indicators, nursing care of patients before and after treatment adherence of two groups were compared.Result:Chronic hepatitis B group’s ANA positive rate was 24.00% (24/100), was significantly higher than the control group’s 4.35% (4/92), the difference was statistically significant (P0.05).After nursing of treatment adherence for patients with chronic hepatitis B was significantly higher than before treatment, the difference was statistically significant (P0.05)。护理后慢性乙肝患者的治疗依从性显著高于护理前,差异有统计学意义(P<0.05)。结论:慢性乙肝患者血清中存在较多抗体,但ANA及肝病相关抗体测定对于预测患者预后和转归具有较大的意义,其可能是导致乙肝后肝硬化的一种病因。同时,为患者制定护理对策并施以护理能增加治疗依从性,值得临床重视。

  11. Establishment of a cell model for screening antibody drugs against rheumatoid arthritis with ADCC and CDC. (United States)

    Yan, Li; Hu, Rui; Tu, Song; Cheng, Wen-Jun; Zheng, Qiong; Wang, Jun-Wen; Kan, Wu-Sheng; Ren, Yi-Jun


    TNFα played a dominant role in the development and progression of rheumatoid arthritis (RA). Clinical trials proved the efficacies of anti-TNFα agents for curing RA. However, most researchers were concentrating on their abilities of neutralizing TNFα, the potencies of different anti-TNFα agents varied a lot due to the antibody-dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC). For better understanding and differentiating the potentiality of various candidate anti-TNF reagents at the stage of new drug research and development, present study established a cell model expressing the transmembrane TNFα for usage in in vitro ADCC or CDC assay, meanwhile, the assay protocol described here could provide guidelines for screening macromolecular antibody drugs. A stable cell subline bearing transmembrane TNFα was first established by conventional transfection method, the expression of transmembrane TNFα was approved by flow cytometer, and the performance of the stable subline in ADCC and CDC assay was evaluated, using human peripheral blood mononuclear cells as effector cells, and Adalimumab as the anti-TNFα reagent. The stable cell subline demonstrated high level of surface expression of transmembrane TNFα, and Adalimumab exerted both ADCC and CDC effects on this cell model. In conclusion, the stable cell line we established in present research could be used in ADCC or CDC assay for screening antibody drugs, which would provide in-depth understanding of the potencies of candidate antibody drugs in addition to the traditional TNFα neutralizing assay.

  12. To screen or not to screen? Celiac antibodies in liver diseases (United States)

    Narciso-Schiavon, Janaína Luz; Schiavon, Leonardo Lucca


    Celiac disease (CD) is a systemic immune-mediated disorder triggered by dietary gluten in genetically predisposed individuals. The typical symptoms are anemia, diarrhea, fatigue, weight loss, and abdominal pain. CD has been reported in patients with primary sclerosing cholangitis, primary biliary cholangitis, autoimmune hepatitis, aminotransferase elevations, nonalcoholic fatty liver disease, hepatitis B, hepatitis C, portal hypertension and liver cirrhosis. We evaluate recommendations for active screening for CD in patients with liver diseases, and the effect of a gluten-free diet in these different settings. Active screening for CD is recommended in patients with liver diseases, particularly in those with autoimmune disorders, steatosis in the absence of metabolic syndrome, noncirrhotic intrahepatic portal hypertension, cryptogenic cirrhosis, and in the context of liver transplantation. In hepatitis C, diagnosis of CD can be important as a relative contraindication to interferon use. Gluten-free diet ameliorates the symptoms associated with CD; however, the associated liver disease may improve, remain the same, or progress. PMID:28223722

  13. Relationship between Human Immunoglobulin and Complement andthe Positive Results of Different Items in the Anti-extractable Nuclear Antigen with the Titer of Antinuclear Antibodies%抗核抗体滴度与IgG,C3,C4水平及抗ENA抗体结果的相关性

    Institute of Scientific and Technical Information of China (English)

    舒颖; 张平安; 魏新素; 叶芳丽; 周心房


    Objective To explore the relationship between immunoglobulin G(IgG) and complement 3 (C3) and C4 with the titer of antinuclear antibodies (ANA) ,and to investigate the correlation between the positive results of different items in the anti-extractable nuclear antigen (anti-ENA) with the titer of ANA.Methods The author analysed the results of autoimmune detection of 1813 patients from May 2010 to June 2012.ANA were tested by indirect immunofluorescence method,anti-ENA antibody were tested by ELISA method,IgG and C3,C4 were determined by immunoturbidimetry.According to the different titers,the patients were divided into several groups,the titer of ANA=1:100 as group l,the titer of ANA=1: 160 as group 2,the titer of ANA=1:320 as group 3,the titer of ANA=1:640 as group 4,the titer of ANA=1:1 280 as group 5,the titer of ANA=1:3 200 as group 6, the titer of ANA>3 200 as group 7,120 cases randomly selected from ANA<1:100 group as group 8(negative group).The change of IgG and complement as well as the positive rates of the specific antibodies of anti-ENA were analyzed by SPSS17.0 software.Results When the titer of ANA≥1:100,the levels of IgG were obviously higher than that in the negative group (F=31.19, P<0.05).When the titer of ANA≥1:320, the levels of C3 and C4 were lower than that in the negative group(F=42.15,9.57,P<0.05).Overall anti-ENA positive rates increased significantly with the titer of ANA (χ2=86.67, P<0.05) ,and the positive rates of various antibodies were different.Anti-SSA and anti-SSB were the most frequent at ANA titers of 1 : 100 to 1 : 3 200,while anti-U1RNP were the most frequent at ANA titers of 1:3 200.Conclusion IgG and C3,C4 were related to the titer of ANA.The positive rates of the overall anti-ENA and some specific antibodies of anti-ENA were related to the titer of ANA.The clinician should pay attention to the value of ANA titer in autoimmune disease diagnosis.%目的 探讨抗核抗体(antinuclear antibodies,ANA)滴度与IgG,C3和C4的

  14. Analysis of Positive Human Immunodeifciency Virus (1+2) Antibodies in Preliminar y Screening:A Report of 394 Cases

    Institute of Scientific and Technical Information of China (English)

    NI Fang; LIU Yan-yan; MA Cai-yun; WU Zhi-qi; XU Hua-guo; JIANG Li


    Objective:To comprehend the characteristics of patients with positive human immunodeifciency virus (HIV) (1+2) antibodies in the preliminary screening so as to provide basis for local HIV screening and prevention. Methods:Enzyme-linked immunosorbent assay (ELISA) was used to detect serum HIV (1 +2) antibodies in the preliminary screening, after which the positive serum was sent to acquired immune deficiency syndrome (AIDS) confirmatory laboratory to be confirmed with western blotting method. Clinical data of patients with positive HIV antibodies in the preliminary screening in the outpatients and inpatients from 2006 to 2013 were collected and analyzed. Results:A total of 394 patients with positive serum HIV antibodies were screened initially, in which 214 were confirmed positive HIV, 13 were not certain and another 167 were negative. Patients with positive serum HIV antibodies in the preliminary screening were increased from 9 cases in 2006 to 94 cases in 2013, in which those conifrmed with positive HIV increased from 5 to 49. Patients with positive serum HIV antibodies in the preliminary screening and those conifrmed increased annually. In addition, patients confirmed with positive serum HIV antibodies were mainly males and aged 20 ~ 49 years, distributing in Departments of Infections, Respiratory, Dermatology, Hematology and Emergency, whereas those confirmed with negative HIV were mainly females and aged >20 years, distributing in Departments of Hematology, Maternity and Emergency as well as Reproductive Center. Conclusion: HIV infection is in low level with characteristics of annually increasing infection rate, male-orientated and wide-spread distribution in variuos departments, etc., knowing which will help guide the clinical practices and carry out the local HIV screening and prevention by relevant departments.

  15. Analysis of the results of antinuclear antibody and anti-ENA antibodies for 297 patients of autoimmune diseases with IIF and biochip technology%297例自身免疫性疾病ANA和抗ENA抗体联合检测分析

    Institute of Scientific and Technical Information of China (English)



    Objective To analyze the clinical features and significance of the samples which the expression of ANA is positive and the expression of anti‐ENA antibody is negative .Methods Detect ANA and anti‐ENA antibody of all the 297 samples by indi‐rect immunofluorescence method and biological chip technology ,repectively .And analyze the fluorescence karyotype of ANA posi‐tive specimens with double‐blind method .Screen the samples which anti‐ENA antibody express negative from ANA positive sam‐ples .Compare and analyze the fluorescent karyotype .Results Of all the 297 samples ,74 cases were positive for ANA ,the positive rate was 24 .9% (74/297) .The majority of patterns were speckled pattern (43 cases ,58 .1% ) ,cytoplasmic granular pattern (9 ca‐ses ,12 .2% ) ,and antibody to centromere antigen(9 cases ,12 .2% ) ,respectively .In 74 cases of ANA positive specimens ,ENA anti‐body the negative rate of ENA antibody is 17 .6% (13/74) .Of the 13 cases ,the most frequently occurred pattern was the speckled pattern(11 cases ,84 .6% ) ,the rate of cytoplasmic granular pattern and antibody to centromere antigen was 7 .7% (1/13) respec‐tively .Conclusion In the serum samples which express ANA positive and ENA antibody negative ,the rate of speckled pattern is significantly higher than the other patterns ,and far more than the rate of speckled pattern in the samples which express ENA anti‐body and ANA positive (52 .5% ) (chi‐square= 5 .018 ,P< 0 .05) ,with statistical significance .We consider the speckled pattern samples be in favour of discovering new antibodys of this kind of serum sample .%目的:分析其中ANA表达阳性而抗ENA抗体表达阴性的样本的检验学特征及意义。方法对297例样本运用间接免疫荧光法检测ANA ,生物芯片技术检测抗ENA抗体,并采用双盲法分析ANA阳性标本的荧光核型。从已知的ANA阳性患者血清中筛选其ENA表达均为阴性的患者血清,比对其荧光

  16. Tumour auto-antibody screening: performance of protein microarrays using SEREX derived antigens

    Directory of Open Access Journals (Sweden)

    Ludwig Nicole


    Full Text Available Abstract Background The simplicity and potential of minimal invasive testing using serum from patients make auto-antibody based biomarkers a very promising tool for use in diagnostics of cancer and auto-immune disease. Although several methods exist for elucidating candidate-protein markers, immobilizing these onto membranes and generating so called macroarrays is of limited use for marker validation. Especially when several hundred samples have to be analysed, microarrays could serve as a good alternative since processing macro membranes is cumbersome and reproducibility of results is moderate. Methods Candidate markers identified by SEREX (serological identification of antigens by recombinant expression cloning screenings of brain and lung tumour were used for macroarray and microarray production. For microarray production recombinant proteins were expressed in E. coli by autoinduction and purified His-tag (histidine-tagged proteins were then used for the production of protein microarrays. Protein arrays were hybridized with the serum samples from brain and lung tumour patients. Result Methods for the generation of microarrays were successfully established when using antigens derived from membrane-based selection. Signal patterns obtained by microarrays analysis of brain and lung tumour patients' sera were highly reproducible (R = 0.92-0.96. This provides the technical foundation for diagnostic applications on the basis of auto-antibody patterns. In this limited test set, the assay provided high reproducibility and a broad dynamic range to classify all brain and lung samples correctly. Conclusion Protein microarray is an efficient means for auto-antibody-based detection when using SEREX-derived clones expressing antigenic proteins. Protein microarrays are preferred to macroarrays due to the easier handling and the high reproducibility of auto-antibody testing. Especially when using only a few microliters of patient samples protein microarrays

  17. High throughput screening for antibody induced complement-dependent cytotoxicity in early antibody discovery using homogeneous macroconfocal fluorescence imaging

    NARCIS (Netherlands)

    Gerritsen, Arnout F.; Bosch, Martijn; de Weers, Michel; van de Winkel, Jan G. J.; Parren, Paul W. H. I.


    Complement-dependent cytotoxicity (CDC) represents an important Fc-mediated effector function of antibodies and is a quality often sought in candidates for therapeutic antibody development in cancer. Antibodies inducing potent CDC are relatively rare as the ability to induce CDC is strongly dependen

  18. Performance characteristics of two automated solid-phase red cell adherence systems for pretransfusion antibody screening: a cautionary tale. (United States)

    Quillen, K; Caron, J; Murphy, K


    Out institution has implemented two instruments, the Galileo and the Echo, that use different solid-phase red cell adherence assays for antibody screening in pretransfusion compatibility testing.During the initial implementation of these two instruments, we noticed very different problems: falsely positive results on the Galileo, and falsely negative results and lack of reproducibility on the Echo. Comparison of falsely positive antibody screen results from approximately equivalent numbers of samples run on the Galileo and samples tested by standard manual tube technique using low-ionic-strength saline enhancement showed a false-positive rate of 1.4 percent on the Galileo (defined as a positive screen with a negative panel). Testing using the Echo identified four cases of falsely negative antibody screens, (defined as a negative screen on a patient sample subsequently shown to be positive by the same method). In addition, we note a lack of reproducibility on the Echo, which emphasizes the importance of replicate testing during validation of automated antibody screening platforms.

  19. Antinuclear Antibodies in Asthma Patients- A Special Asthma Phenotype?

    Directory of Open Access Journals (Sweden)

    Agache Ioana


    Full Text Available Several studies reported the appearance of asthma and autoimmune conditions in the same patient, but the clinical significance of this association was not yet assessed. One hundred asthmatic patients were observed for one year evolution with death, severe exacerbations, intake of > 1000 micrograms of beclometasone or equivalent (high ICS and FEV1 decline >100 ml, in relation with ANA (ELISA, sputum and blood eosinophilia (EO, NSAID intolerance, BMI >25, chronic rhinosinusitis, smoking status and FEV1 After 1 year of observation, there were 5 deaths, 28 severe asthma exacerbations requiring hospitalisations, 24 cases requiring high inhaled corticosteroid intake, and 19 patients with fast FEV1 decline (>100 ml/year. Multiple regression analysis pointed out several different independent risk factors for severe asthma evolution: for death presence of ANA (P=0.037, NSAID intolerance (P100 ml ANA (P=0.006, sputum EO (P=0.037, BMI>25 (P=0.046 and NSAID intolerance (P=0.017The presence of ANA is an independent risk factor in asthma for evolution with death, severe exacerbations, high inhaled corticosteroid intake and FEV1 decline >100 ml.

  20. Evaluation of five screening tests licensed in Argentina for detection of hepatitis C virus antibodies

    Directory of Open Access Journals (Sweden)

    Viviana Ré


    Full Text Available This study was conducted to compare among the most recent generation of five screening tests licensed in Argentina, in order to evaluate which of the tests has the best sensitivity for detection of antibodies against hepatitis C virus (HCV. The tests analyzed were: Detect-HCV™ (3.0 Biochem ImmunoSystems, Canada; Hepatitis C EIA Wiener Lab., Argentina; Equipar HCV Ab, Italy; Murex HCV 4.0, UK and Serodia-HCV particles agglutination test, Japan. The results obtained showed high discrepancy between the different kits used and show that some of the tests assessed have a low sensitivity for anti-HCV detection in both chronic infections and early seroconversion, and indicate that among the commercially available kits in Argentina, Murex HCV 4.0 (UK and Serodia-HCV particles agglutination test (Japan have the best sensitivity for HCV screening. Although the sensitivity of the assays is the first parameter to be considered for blood screening, more studies should be carried out to assess the specificity of such assays.

  1. Antibody (United States)

    An antibody is a protein produced by the body's immune system when it detects harmful substances, called antigens. Examples ... microorganisms (bacteria, fungi, parasites, and viruses) and chemicals. Antibodies may be produced when the immune system mistakenly ...

  2. Antibody screening tests variably overestimate the prevalence of hepatitis C virus infection among HIV-infected adults in Ghana. (United States)

    King, S; Adjei-Asante, K; Appiah, L; Adinku, D; Beloukas, A; Atkins, M; Sarfo, S F; Chadwick, D; Phillips, R O; Geretti, A M


    HIV coinfection with HCV has been poorly studied in sub-Saharan Africa, and the reliability of available seroprevalence estimates remains uncertain. The study aim was to determine HCV RNA prevalence in HIV-infected subjects receiving care in Kumasi, Ghana, and relate the findings to HCV antibody detection. From a population of 1520 HIV-infected adults, all HBsAg-positive subjects (n = 236) and a random subset of HBsAg-negative subject (n = 172) were screened for HCV RNA using pooled plasma; positive samples were genotyped by core and NS5B sequencing. HCV antibodies were detected by three commercial screening assays and confirmed by the line immunoassay. HCV RNA was detected in 4/408 subjects (1.0%, 95% confidence interval 0.0-1.9%), comprising 3/236 (1.3%; 0.0-2.8%) HBsAg-positive and 1/172 (0.6%; 0.0-1.8%) HBsAg-negative subjects. HCV RNA-positive subjects showed reactivity in all three antibody screening assays. Among HCV RNA-negative subjects, 5/67 (7.5%), 5/67 (7.5%) and 19/67 (28.4%) showed antibody reactivity by each screening assay, respectively, including two (3.0%) with reactivity by all three assays. Only one sample (1.5%) had confirmed antibody reactivity by line immunoassay indicating past HCV infection. HCV-positive subjects (three males, two females) were aged 30-46 years, by questionnaire-based interview reported surgical procedures and blood transfusion as risk factors for infection. HCV genotypes were 2 (subtypes 2j, 2l, 2k/unassigned) and 1 (subtype unassigned). Without further testing, HCV antibody screening assays variably overestimated HCV prevalence among HIV-infected subjects in Ghana. These findings inform the interpretation of previous seroprevalence estimates based upon screening assays alone.

  3. Bioreactor process parameter screening utilizing a Plackett-Burman design for a model monoclonal antibody. (United States)

    Agarabi, Cyrus D; Schiel, John E; Lute, Scott C; Chavez, Brittany K; Boyne, Michael T; Brorson, Kurt A; Khan, Mansoor A; Read, Erik K


    Consistent high-quality antibody yield is a key goal for cell culture bioprocessing. This endpoint is typically achieved in commercial settings through product and process engineering of bioreactor parameters during development. When the process is complex and not optimized, small changes in composition and control may yield a finished product of less desirable quality. Therefore, changes proposed to currently validated processes usually require justification and are reported to the US FDA for approval. Recently, design-of-experiments-based approaches have been explored to rapidly and efficiently achieve this goal of optimized yield with a better understanding of product and process variables that affect a product's critical quality attributes. Here, we present a laboratory-scale model culture where we apply a Plackett-Burman screening design to parallel cultures to study the main effects of 11 process variables. This exercise allowed us to determine the relative importance of these variables and identify the most important factors to be further optimized in order to control both desirable and undesirable glycan profiles. We found engineering changes relating to culture temperature and nonessential amino acid supplementation significantly impacted glycan profiles associated with fucosylation, β-galactosylation, and sialylation. All of these are important for monoclonal antibody product quality.

  4. Screening Response to Hepatitis C Virus Antibodies among Diabetic Patients Attending UITH Nigeria

    Directory of Open Access Journals (Sweden)

    J.A. Ndako


    Full Text Available Epidemiological studies have suggested that hepatitis C virus (HCV infection is a risk factor for the development of diabetes mellitus (DM type 2; hence this study was carried out to investigate the prevalence of hepatitis C Virus (HCV among diabetic patients attending University of Ilorin Teaching Hospital (UITH. A total number of one hundred and eighty diabetic patients made up of seventy five males (41.7% and one hundred and five females (58.3% were recruited for the study. Structured questionnaire on demographic data and risk factors for HCV was administered to the participants. The sera of all the subjects were assayed for antibodies to HCV using a fourth generation enzyme linked immunosorbent Assay [Forte Diagnostic Limited], All the sera were tested for antibodies to Hepatitis C virus by ELISA method. The overall prevalence of HCV infection among diabetic patients was 5.0%. Of the nine participants’ positive for anti-HCV antibodies, three were males (1.7% while six were females (3.3%. Although more female attended the diabetic clinic, there was no significant difference between HCV infection and sex (p = 0.603; p>0.05. The prevalence of HCV infection was highest in the 40-60 age categories. Two of the seropositive individuals had elevated transaminases, with one of the two being an alcoholic consumer. Type 2 diabetes is a debilitating disease condition especially in individuals above 30 years of age, with these scenario it has become very vital for screening exercise to be carried out so as to determine the prevalence rate of HCV among diabetic patient.

  5. New antibody and immunoassay pretreatment strategy to screen polychlorinated biphenyls in Korean transformer oil. (United States)

    Terakado, Shingo; Ohmura, Naoya; Park, Seok-Un; Lee, Seung-Min; Glass, Thomas R


    Development and modifications are described that expand the application of an immunoassay from the detection of Kanechlors (Japanese technical PCBs mixtures) to the detection of Aroclors (U. S. technical PCB mixtures, used in Korea) in contaminated Korean transformer oil. The first necessary modification was the development of a new antibody with a reactivity profile favorable for Aroclors. The second modification was the addition of a second column to the solid-phase extraction method to reduce assay interference caused by the Korean oil matrix. The matrix interference is suspected to be caused by the presence of synthetic oils (or similar materials) present as contaminants. The modified assay was validated by comparison to high-resolution gas chromatography/high-resolution mass spectrometry analysis, and was shown to be tolerant of up to 10% of several common synthetic insulating oils. Finally the screening performance of the modified assay was evaluated using 500 used transformer oil samples of Korean origin, and was shown to have good performance in terms of false positive and false negative rates. This report provides evidence for the first establishment of immunoassay screening for Aroclor based PCB contamination in Korean transformer oil.

  6. REAL-Select: full-length antibody display and library screening by surface capture on yeast cells. (United States)

    Rhiel, Laura; Krah, Simon; Günther, Ralf; Becker, Stefan; Kolmar, Harald; Hock, Björn


    We describe a novel approach named REAL-Select for the non-covalent display of IgG-molecules on the surface of yeast cells for the purpose of antibody engineering and selection. It relies on the capture of secreted native full-length antibodies on the cell surface via binding to an externally immobilized ZZ domain, which tightly binds antibody Fc. It is beneficial for high-throughput screening of yeast-displayed IgG-libraries during antibody discovery and development. In a model experiment, antibody-displaying yeast cells were isolated from a 1:1,000,000 mixture with control cells confirming the maintenance of genotype-phenotype linkage. Antibodies with improved binding characteristics were obtained by affinity maturation using REAL-Select, demonstrating the ability of this system to display antibodies in their native form and to detect subtle changes in affinity by flow cytometry. The biotinylation of the cell surface followed by functionalization with a streptavidin-ZZ fusion protein is an approach that is independent of the genetic background of the antibody-producing host and therefore can be expected to be compatible with other eukaryotic expression hosts such as P. pastoris or mammalian cells.

  7. REAL-Select: full-length antibody display and library screening by surface capture on yeast cells.

    Directory of Open Access Journals (Sweden)

    Laura Rhiel

    Full Text Available We describe a novel approach named REAL-Select for the non-covalent display of IgG-molecules on the surface of yeast cells for the purpose of antibody engineering and selection. It relies on the capture of secreted native full-length antibodies on the cell surface via binding to an externally immobilized ZZ domain, which tightly binds antibody Fc. It is beneficial for high-throughput screening of yeast-displayed IgG-libraries during antibody discovery and development. In a model experiment, antibody-displaying yeast cells were isolated from a 1:1,000,000 mixture with control cells confirming the maintenance of genotype-phenotype linkage. Antibodies with improved binding characteristics were obtained by affinity maturation using REAL-Select, demonstrating the ability of this system to display antibodies in their native form and to detect subtle changes in affinity by flow cytometry. The biotinylation of the cell surface followed by functionalization with a streptavidin-ZZ fusion protein is an approach that is independent of the genetic background of the antibody-producing host and therefore can be expected to be compatible with other eukaryotic expression hosts such as P. pastoris or mammalian cells.

  8. The clinical study on the risk of permanent hair dye use for connective tissue disease with positive antinuclear antibodies%永久性染发剂与抗核抗体阳性结缔组织病发病风险的临床研究

    Institute of Scientific and Technical Information of China (English)

    宫玉芳; 魏巧凤; 王秀花; 吕明; 李兴福


    Objective To investigate the risk of permanent hair dye use for connective tissue disease ( CTD ) with positive antinuclear antibodies ( ANA ).Methods We conducted a population-based case-control study to analyze 440 CTD patients from the department of our hospital.Four hundreds and forty patients' relatives or friends who were free from any CTD were selected as controls.Patients and controls were matched in sex and age.A face-to-face interview was adopted to finish questionnaires about subjects' demographic data and the use of permanent hair dye.Data about the duration,frequency,times of usage and allergy related to hair dye were collected.Medical history was also reviewed,such as the diagnosis,disease course,primary symptoms,serum tests and organ involvements.Statistical analysis was performed by SPSS 17.0 software using x2 test and Logistic regression.Results The association between the use of permanent hair dye and CTD did not reach statistical significance (OR=1.282,95%CI 0.966-1.700,P=0.085 ),when the relationship of hair dye use with systemic lupus erythematosus (SLE) was tested,the same result (OR=1.092,95%CI 0.795-1.500,P=0.587 ) could be revealed.Conclusion This preliminary study has shown that the use of permanent hair dye couldn't induce the occurrence of CTD.%目的 探讨使用永久性染发剂与抗核抗体阳性结缔组织病(CTD)发病的风险.方法 采用病例对照研究,对440例CTD患者和性别、年龄相匹配的440名健康的患者家属或朋友进行比较分析.采用面对面的交谈,以调查问卷的方式记录研究对象的基本信息和染发信息(包括染发时间、染发频率、染发次数及与染发相关的过敏).记录病例组的现病史,包括疾病的诊断、病程、初发症状、各脏器受累情况及各项实验室检查.应用SPSS 17.0软件进行x2检验和Logistic回归分析.结果 使用永久性染发剂与CTD发病无明确因果关系[比值比(OR)=1.282,95%可信区间(CI)0.966~1

  9. 噬菌体抗体库筛选技术%Screening Isolates from Antibody Phage-display Libraries

    Institute of Scientific and Technical Information of China (English)

    高鹏; 胡立勇; 钱钰; 徐晨


    For almost 20 years, phage display antibody (Ab) libraries screening have been widely used in antibody screen, clinical diagnostic, treatment of disease and basic research areas. Rapid identification of the autofit monoclonal antibody, and reliable and efficient data management and analysis are very important in the application of this technology. This review described reported methods for high-throughput screening of antibody phage-display libraries and further more gave a brief introduction to data management.%在过去的20年中,噬菌体抗体库筛选技术被广泛的应用于抗体筛选、疾病治疗,临床诊断以及基础研究之中.在该技术的应用过程中,快速有效的筛选出最适合的单克隆抗体并进行可靠和高效的数据管理和分析是十分重要的.文中总结了近年来噬菌体抗体库的高通量筛选方法并且对数据管理做了简要介绍.

  10. Establishment and application of biotin-avidin-system-time-resolved fluoroimmunoassay for the quantitive detection of antinuclear antibody-Ig%生物素-亲合素系统-时间分辨荧光免疫分析法检测抗核抗体方法学的建立和临床应用

    Institute of Scientific and Technical Information of China (English)

    胡志刚; 刘洁; 陈国千; 邹耀红; 俞蕾


    Objective To establish an analytical method with high sensitivity and wide range in biotin-avidin and time-resolved fluoroimmunoassay system for the quantitative detection of antinuclear antibody (ANA)-Ig (GAM).Methods ANA in standard preparation or sample was combined with solid nuclear antigen,biotinylation antibody and the europium(Eu3+)-labelled avidin to form the compounds of solid nuclear antigen-antibody-biotinylation antibody-SA-Eu3+.The fluorescence enhancement fluid was added to dis-sociate the Eu3+,the content of ANA was directly proportional to fluorescence intensity,the BAS-TRFLA was established for the quantitative detection of ANA-Ig (GAM).The sensitivity,specificity,reliability and range of detection were evaluated.Semm of 50 blood donor,105 patients with systemic lupus erythematosus (SLE),109 patients with rheumatoid arthritis (RA),25 patients with PBC,29 patients with Sj(o)gren's syndrome (SS),23 patients with scleroderma,25 patients with MCTD were included in this study.The positive rate was calculated.Results The inner-group difference between high,medium and low densities mixture serum was 3.13%,3.74% and 5.76% and the inter-group precision rate was 5.31%,6.25% and 7.46% in BAS-TRFLA.The sensitivity of TRFIA was better than that of the ELISA method.The low detection limit was 2.24 U/ml.The mean recovery rate was 100.74%.The results measured by the TRFIA and ELISA methods were closely correlated (R2=0.978).The positive rate of blood donor,and patient with SLE,RA,PBC,SS,scleroderma and MCTD were 0,100%,23%,96%,38%,91%,92% respectively.When compared with ELISA,the detection range of TRFIA was wider,and stability was better.Conclusion BAS-TRFIA is a stable method for detection of ANA with high sensitive and wide range of detection.It is important for the early diagnosis of autoimmune disease and monitoring the treatment efficacy of autoimmune disease.And this method may be widely used in clinical laboratories in the future

  11. HIV抗体初筛实验室的管理%Management of HIV Antibody Primary Screening Lab

    Institute of Scientific and Technical Information of China (English)

    吕永强; 徐宏亮


    从实验室建设、人员配备与培训、仪器管理、文件管理、试剂管理、质量控制、安全防护等方面对HIV抗体初筛实验室的管理进行总结,指出,规范化的实验室管理是加强HIV抗体初筛实验室质量控制的有效手段.%Relative problems in the management of HIV antibody primary screening lab are discussed, including laborato-ry building, staffing and training, equipment management, document management, reagent management, quality control and security protection. It is pointed out that the standardized laboratory management is an important part of the HIV antibody primary screening lab's quality Control.

  12. Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Daohong [Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062 (China); Li Peiwu [Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062 (China); Quality Inspection and Test Center for Oilseeds and Products of Ministry of Agriculture, Wuhan 430062 (China)], E-mail:; Zhang Qi; Zhang Wen; Huang Yanling; Ding Xiaoxia; Jiang Jun [Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062 (China); Quality Inspection and Test Center for Oilseeds and Products of Ministry of Agriculture, Wuhan 430062 (China)


    Monoclonal antibodies (McAbs) cross-reactive with four major aflatoxins were achieved using a modified two-step screening procedure. The first step was twice modified indirect enzyme-linked immunosorbent assay (ELISA) and resulted in positive hybridomas and hapten-specific antibodies. The modified indirect competitive ELISA (ciELISA) was the second step, in which the competition incubation time was decreased to 30 min, aflatoxin B{sub 1}, B{sub 2}, G{sub 1} and G{sub 2} were all used as competitors, the concentrations of four aflatoxins were gradiently decreased in each screening. 2-3 subclonings were performed after every modified fusion and resulted in eight hybridomas that secreted antibodies with good cross-reactivity and high affinity to four aflatoxins. Five McAbs were chosen for further analysis. Of the five, two antibodies had similar reaction efficiency with aflatoxin B{sub 1}, B{sub 2} and G{sub 1} but showed a weak cross-reaction to G{sub 2}. Another two had almost identical reaction capability with four aflatoxins. One clone 1C11 exhibited the highest sensitivity for all four aflatoxins. The concentrations of aflatoxin B{sub 1}, B{sub 2}, G{sub 1} and G{sub 2} at 50% inhibition for 1C11 were 1.2, 1.3, 2.2 and 18.0 pg mL{sup -1} respectively. This is the most sensitive for all four major aflatoxins described so far. The results indicated that the modified two-step screening procedure had superiority and these antibodies could be used for simultaneous analysis of total aflatoxins.

  13. Understanding ForteBio's Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant. (United States)

    Yu, Yao; Mitchell, Scott; Lynaugh, Heather; Brown, Michael; Nobrega, R Paul; Zhi, Xiaoyong; Sun, Tingwan; Caffry, Isabelle; Cao, Yuan; Yang, Rong; Burnina, Irina; Xu, Yingda; Estep, Patricia


    Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts.

  14. Screening of Human Antibody Fab Fragment against HBsAg and the Construction of its dsFv Form

    Directory of Open Access Journals (Sweden)

    Leili Jia, Jiyun Yu, Hongbin Song, Xuelin Liu, Weina Ma, Yuanyong Xu, Chuanfu Zhang, Shicun Dong, Qiao Li


    Full Text Available The objective of this study was to pursue the techniques involving the screening of the human antibody Fab fragment against hepatitis B virus surface antigen (HBsAg and the construction of its disulfide-stabilized Fv fragment (dsFv. The phage antibody Fab fragments against HBsAg were screened from the human combinatorial immunoglobulin library. Sequence analysis revealed that its heavy chain gene was complete, but the light chain gene was lost. To improve the affinity of the antibody by chain shuffling, a human antibody light chain gene repertoire was generated by reverse transcriptase-polymerase chain reaction (RT-PCR from the human peripheral blood lymphocytes. A phage antibody sub-library was then constructed by inserting the light chain gene repertoire into the phagmid that contained the Fd gene. Five clones with appreciably higher absorbance than that of the original clone were obtained, which indicated that the affinity of the light chain-shuffled phage antibodies was improved. Then, the mutated genes of dsFv against HBsAg were constructed by using PCR-based point mutagenesis method. Purified VH and VL proteins were folded into a 25-kDa protein, designated as anti-HBsAg dsFv. ELISA and competition ELISA revealed that the dsFv maintained the specificity of the Fab by binding to HBsAg, even through with a lower binding activity. These results have facilitated the undertaking of further functional analyses of the constructed dsFv, and may therefore provide an improved technique for the production and application of dsFvs against HBsAg.

  15. Retrospective study on effect of antinuclear antibody, extractable nuclear antigen and antineutrophil cytoplasmic antibody on clinical manifestation and cerebrospinal fluid characteristics of neuromyelitis optica%血清抗核抗体、可提取性核抗原、抗中性粒细胞胞质抗体对视神经脊髓炎临床特点及急性复发期脑脊液特征的影响

    Institute of Scientific and Technical Information of China (English)

    刘彩燕; 徐雁; 崔丽英; 陈琳


    Objective To explore the effect of antinuclear antibody ( ANA+dsDNA),extractable nuclear antigen (ENA) and antineutrophil cytoplasmic antibody (ANCA) on the clinical manifestation and cerebrospinal fluid characteristics of neuromyelitis optica (NMO).Methods All 41 patients with NMO in PUMC hospital from 1985 to 2009 were retrospectively reviewed.All patients underwent examination of serum ANA+dsDNA,ENA and ANCA.Fourteen positive-autoantibody patients were compared with 27negative-autoantibody patients in gender,onset age,duration,relapse ratio,first demyelination event,the extent of optic neuritis and myelitis,EDSS,CSF protein,WBC,Oligoclonal band, 24 hours IgG index and myelin basic protein.Results The 14 NMO patients (34.1%) had positive non-organ-specific antibodies.NMO patients who had negative autoantibodies were compared with NMO patients with positive autoantibodies with significantly higher EDSS (the EDSS score were 4.5 and 2.5 respectively,U=92.5,P=0.008),more complete damage of spinal cord (3/14 vs 0/27, x2=6.736, P=0.0095) and tended to have higher visual Function Scale in remitting phase.There was no significant difference on the gender,onset age,duration,relapse ratio,first demyelination event.The positive-autoantibody patients had higher CSF WBC (2.0 vs 0,U=68.0,P=0.007) and tended to have lower 24 hours IgG index (-8.663 vs 0.163,U=30.0,P=0.053).There was no significant difference in CSF protein,MBP and OB.Conclusion NMO patients with positive autoantibodies have more severe intrathecal autoimmune inflammatory and disability,so they might need more intensive treatment.%目的 探讨血清抗核抗体(ANA+dsDNA)、可提取性核抗原(ENA)、抗中性粒细胞胞质抗体(ANCA)对视神经脊髓炎(NMO)临床特点及急性复发期脑脊液特征的影响.方法 回顾分析1985-2009年北京协和医院收治的临床诊断为NMO的41例患者,均进行血清ANA+dsDNA、ENA和ANCA检查,比较其中14例自身抗体阳性的NMO患者与27例自身

  16. Vi serology in screening of typhoid carriers: improved specificity by detection of Vi antibodies by counterimmunoelectrophoresis.


    Chau, P. Y.; Tsang, R S


    A purified soluble Vi antigen was used in counterimmunoelectrophoresis (CIE), passive haemagglutination (HA) and solid phase radio-immunoassay (SPRIA) for detection of serum Vi antibody. Serum Vi antibody was detected in 13 of 14 chronic typhoid carriers by both CIE and HA. SPRIA studies showed that Vi antibodies in sera of these carriers were mainly of the IgG class. Successful treatment with amoxycillin in one carrier, who initially showed a positive Vi test, resulted in a reversion in 6 mo...

  17. Clinical characteristics of children with positive anti-SSA/SSB antibodies. (United States)

    Chen, Pei-Hsuan; Yang, Yao-Hsu; Lin, Yu-Tsan; Lee, Jyh-Hong; Wang, Li-Chieh; Yu, Hsin-Hui; Chiang, Bor-Luen


    This study aimed to characterize the manifestations of clinical symptoms and signs, primary rheumatic diseases, and other autoantibodies in pediatric patients with positive anti-SSA and/or anti-SSB antibodies. Subjects under age 18 with positive anti-SSA and/or anti-SSB antibodies were screened and enrolled in a tertiary hospital in Taiwan. Data were collected via medical records,including age, gender, onset of the primary rheumatic disease, clinical symptoms and signs, and the medication used. Schirmer test for Sjögren's syndrome (SS) screening was performed in all enrolled patients. Among twenty enrolled subjects, seventeen of them had systemic lupus erythematosus; four of them were diagnosed as SS with positive Schirmer test. In addition to antinuclear antibodies and anti-DNA antibodies, other common autoantibodies were anti-RNP antibodies (50 %) and anti-Sm antibodies(30 %). The most common symptoms were arthritis (60 %)followed by malar rash (40 %). In conclusion, we observed that a low proportion of childhood SS (4/20) exists in our patients with positive SSA and/or anti-SSB antibodies. It is suggested that clinicians should focus more on the clinical symptoms in these patients, rather than undertaking invasive diagnostic interventions to rule out Sjögren's syndrome.

  18. Screening for uveitis in juvenile chronic arthritis. (United States)

    Kanski, J J


    Three hundred and fifteen patients with anterior uveitis associated with juvenile chronic arthritis (JCA) were studied in order to identify the various risk factors for uveitis. Girls were more susceptible to uveitis than boys by a ratio of 3:1. In 94% of cases the uveitis was diagnosed after the development of arthritis. The risk of uveitis was small after seven or more years had elapsed from the onset of arthritis. Patients with pauciarticular onset JCA had the highest risk of uveitis and systemic onset patients the least risk. The presence of circulating antinuclear antibody was also an important marker for an increased risk of uveitis. A regimen for routine screening of patients is suggested.

  19. Antibody levels after regular childhood vaccinations in the immunological screening of children with recurrent otitis media.

    NARCIS (Netherlands)

    Wiertsema, S.P.; Sanders, E.A.M.; Veenhoven, R.H.; Heerbeek, N. van; Hof, S. van den; Berbers, G.A.; Rijkers, G.T.


    Recurrent otitis media may be related to defects in specific antibody production, as suggested previously. This might be reflected in lower antibody responses to vaccinations administered in the context of the national childhood vaccination program in children suffering from recurrent otitis media.

  20. Optimizing selection of large animals for antibody production by screening immune response to standard vaccines. (United States)

    Thompson, Mary K; Fridy, Peter C; Keegan, Sarah; Chait, Brian T; Fenyö, David; Rout, Michael P


    Antibodies made in large animals are integral to many biomedical research endeavors. Domesticated herd animals like goats, sheep, donkeys, horses and camelids all offer distinct advantages in antibody production. However, their cost of use is often prohibitive, especially where poor antigen response is commonplace; choosing a non-responsive animal can set a research program back or even prevent experiments from moving forward entirely. Over the course of production of antibodies from llamas, we found that some animals consistently produced a higher humoral antibody response than others, even to highly divergent antigens, as well as to their standard vaccines. Based on our initial data, we propose that these "high level responders" could be pre-selected by checking antibody titers against common vaccines given to domestic farm animals. Thus, time and money can be saved by reducing the chances of getting poor responding animals and minimizing the use of superfluous animals.

  1. An experimental design approach to optimize an amperometric immunoassay on a screen printed electrode for Clostridium tetani antibody determination. (United States)

    Patris, Stéphanie; Vandeput, Marie; Kenfack, Gersonie Momo; Mertens, Dominique; Dejaegher, Bieke; Kauffmann, Jean-Michel


    An immunoassay for the determination of anti-tetani antibodies has been developed using a screen printed electrode (SPE) as solid support for toxoid (antigen) immobilization. The assay was performed in guinea pig serum. The immunoreaction and the subsequent amperometric detection occurred directly onto the SPE surface. The assay consisted of spiking the anti-tetani sample directly onto the toxoid modified SPE, and then a second antibody, i.e. a HRP-labeled anti-immunoglobulin G, was deposited onto the biosensor. Subsequent amperometric detection was realized by spiking 10 µL of a hydroquinone (HQ) solution into 40 µL of buffer solution containing hydrogen peroxide. An experimental design approach was implemented for the optimization of the immunoassay. The variables of interest, such as bovine serum albumin (BSA) concentration, incubation times and labeled antibody dilution, were optimized with the aid of the response surface methodology using a circumscribed central composite design (CCCD). It was observed that two factors exhibited the greatest impact on the response, i.e. the anti-tetani incubation time and the dilution factor of the labeled antibody. It was discovered that in order to maximize the response, the dilution factor should be small, while the anti-tetani antibody incubation time should be long. The BSA concentration and the HRP-anti-IgG incubation had very limited influence. Under the optimized conditions, the immunoassay had a limit of detection of 0.011 IU/mL and a limit of quantification of 0.012 IU/mL. These values were below the protective human antibody limit of 0.06 IU/mL.

  2. Anti-ENA antibody profile in hepatitis C patients undergoing hemodialysis. (United States)

    Batchoun, Raymond G; Al-Najdawi, Malek A; Al-Taamary, Sameh


    Infection with hepatitis C virus (HCV) is increasing all over the world, especially among hemodialysis patients. HCV is one of the major autoantibody inducing viruses, where anti-nuclear antibodies (ANA), anti-smooth muscle antibodies (ASMA), anti-liver kidney microsome antibodies (LKM-1), and rheumatoid factor (RF) have been related to HCV. Few studies have investigated the presence of anti-extractable nuclear antigens (ENA) antibodies in chronic liver diseases, especially in chronic hepatitis C cases, but none investigated its immunostimulation role in hemodialysis units. The aim of the study was to assess the prevalence of HCV among chronic kidney disease- Stage 5 (CKD5) patients undergoing hemodialysis and the prevalence of ENA antibodies among them. Sera of 134 patients with chronic kidney disease undergoing hemodialysis, were screened for HCV antibodies and ENA antibodies profile, using ELISA and Immunoblot technique. 41 HCV-positive blood bank donors were used as controls. Sixty-four (47.7%) of 134 patients undergoing hemodialysis were infected with HCV. Thirty-three (51.6%) of 64 patients with HCV infection undergoing hemodialysis had anti-ENA antibodies: 9 (27.3%) showed anti-SSA antibodies and 22 (66.7%) had anti-SSB antibodies. The prevalence of anti-ENA antibodies was significantly higher in the patients with HCV infection, undergoing hemodialysis, compared with both control groups (hepatitis C-positive blood bank donors and hepatitis C-negative patients undergoing hemodialysis). Seventeen of 33 HCV antibodies-positive males undergoing hemodialysis had anti-ENA antibodies, compared with 16 of 31 females, indicating no sex related difference. This study emphasizes the high prevalence of HCV infection in our hemodialysis patients, comparable to that of other Middle Eastern countries, but higher than Western ones. A strong association was observed between anti-HCV positivity and hemodialysis duration, as well as anti-ENA antibody profile. However, these

  3. [IgA-class antigliadin antibodies in the screening and follow-up of celiac disease patients]. (United States)

    Papadatou, B; Ferretti, F; Colistro, F; Castellucci, G; Lucidi, V; Ricci, S; Cerminara, R; Bella, S; Colombo, A M; Gambarara, M


    IgA antigliadin antibodies (IgA-AGA) have been determined with an enzyme immunoassay in 2.141 pediatric patients. High levels of IgA were found in 98% of 53 celiac patients (1st biopsy), in 81% of 16 celiac patients after gluten challenge, while high levels of these antibodies were not found in 200 patients on gluten-free diet. Moreover high levels of IgA-AGA were found in 29% of 48 patients with normal jejunal biopsy and in 4% of 1.824 patients with gastrointestinal problems other than celiac disease. Our results confirm the data report in literature about the sensibility and the specificity of the IgA-AGA dosage as a screening test for celiac disease, but the possibility of false pathological and false normal values confirms the intestinal biopsy, as the main procedure for the diagnosis of celiac disease.

  4. Immunohistochemical screening for antibodies in recent onset type 1 narcolepsy and after H1N1 vaccination. (United States)

    van der Heide, Astrid; Hegeman-Kleinn, Ingrid M; Peeters, Els; Lammers, Gert J; Fronczek, Rolf


    Narcolepsy type 1 patients typically have undetectable hypocretin-1 levels in the cerebrospinal fluid (CSF), as a result of a selective loss of the hypocretin containing neurons in the hypothalamus. An autoimmune attack targeting hypothalamic hypocretin (orexin) neurons is hypothesised. So far, no direct evidence for an autoimmune attack was found. One of the major limitations of previous studies was that none included patients close to disease onset. We screened serum of 21 narcolepsy type 1 patients close to disease onset (median 11 months), including 8 H1N1 vaccinated patients, for antibodies against hypocretin neurons using immunohistochemistry. No autoantibodies against hypocretin neurons could be detected.

  5. A general strategy for antibody library screening via conversion of transient target binding into permanent reporter deposition. (United States)

    Maaß, Alexander; Heiseler, Tim; Maaß, Franziska; Fritz, Janine; Hofmeyer, Thomas; Glotzbach, Bernhard; Becker, Stefan; Kolmar, Harald


    We report here a generally applicable method for the selective covalent attachment of a reporter molecule to a replicating entity that allows one to obtain specific binders from a single round of library screening. We show that selective biotinylation of phage particles displaying a binder to any given target can be achieved by application of a coupled enzyme reaction on the surface of the target-binding phage particles that includes a peroxidase, an oxidase and a catalase. Due to the covalent linkage of biotin together with the tight and stable interaction of biotin with streptavidin, very stringent wash conditions for removal of nonspecific binders can be applied. The method termed (3)CARD (triple catalytic reporter deposition) was successfully applied to single-round screening of a phage display library of camelid single-domain antibodies against three different target proteins.

  6. Evaluation of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting

    Directory of Open Access Journals (Sweden)

    Ingole N


    Full Text Available Purpose: Integrated counselling and testing centres (ICTC provide counselling and blood testing facilities for HIV diagnosis. Oral fluid tests provide an alternative for people whodo not want blood to be drawn. Also, it avoids the risk of occupational exposure. The goal of this study was to evaluate the utility of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting. Materials and Methods: A cross-sectional study was carried out after ethics committee approval in 250 adult ICTC clients. Blood was collected and tested from these clients for HIV diagnosis as per routine policy and the results were considered as the gold standard. Also, after another written informed consent, oral fluid was collected from the clients and tested for the presence of HIV antibodies. Twenty five clients who had and 25 clients who had not completed their secondary school education (Group A and Group B, respectively were also asked to perform and interpret the test on their own and their findings and experiences were noted. Result: The sensitivity, specificity, PPV and NPV of the oral fluid antibody test were 100%, 98.51%, 94.11% and 100%, respectively. Seventy six percent of clients preferred oral fluid testing. Group B found it difficult to perform the test as compared to Group A and this difference was statistically significant (P ≤ 0.05. Conclusion: Oral fluid testing can be used as a screening test for HIV diagnosis; however, confirmation of reactive results by blood-based tests is a must.

  7. Factors Associated with Participation in HIV Antibody Screening and Results Disclosure. (United States)

    Silvestre, Anthony J.; And Others


    Identified differences among 110 gay and bisexual men who decided whether to be tested for antibodies to human immunodeficiency virus (HIV) and, if so, whether to return for results. Fifty percent refused testing. Of those tested, only 35% returned to obtain test results. Education was significantly and inversely related to being tested and to…

  8. Specificity of mannan antigen and anti-mannan antibody screening in patients with haematological malignancies at risk for fungal infection. (United States)

    Duettmann, Wiebke; Koidl, Christoph; Krause, Robert; Lackner, Gertrude; Woelfler, Albert; Hoenigl, Martin


    Combination of mannan antigen and anti-mannan antibody (Mn/A-Mn) testing has been reported a useful and specific strategy for diagnosis of invasive Candida infections (ICIs). We evaluated Mn/A-Mn as a screening tool in patients with haematological malignancies. This clinical prospective study was performed at the Division of Hematology, Medical University Graz, Austria between July and December 2012. Patients at risk for fungal infection were included into the study and twice weekly screened by Mn/A-Mn testing, yielding 650 samples. Of overall 67 patients 66 had no evidence for ICI. From those, 153/640 serum samples (23.9%) were positive for mannan Ab, and nine (1.4%) for Ag. Most false positive Ab results were observed among 375 samples from patients without haematopoietic stem cell transplantation (34.9% resulted positive). Combined specificity of Mn/A-Mn was 74.8%. Of 10 samples obtained in the single patient with candidemia, five were positive for mannan Ag (from the day of diagnosis up to 40 days after detection of candidemia) and none for Ab. In conclusion, mannan Ab screening yielded a high number of false positive results. While mannan Ag was found to be highly specific and may have potential for diagnostic driven testing, mannan Ab testing cannot be recommended based on our study results.

  9. Anti-enrofloxacin Antibody Production by Using Enrofloxacin-screened HSA as an Immunogen

    Institute of Scientific and Technical Information of China (English)

    LIU Chune; LIN Hong; CAO Limin; JIANG Jie


    A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating enrofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and then used for immunization of BALB/C mice. In enzyme-linked immunosorbent assay (ELISA) and competitive inhibition ELISA experiments, the derived antiserum exhibited high antibody titer (greater than 1: 250 000) as well as varied cross-reactivity (from 97.8% to 161.7%) to three analogs of EF belonging to fluoroquinolones family. But over the concentration range studied, no significant cross-reactivity was observed to other group of antibiotics (chloramphenicol, oxytetracycline, sulphamethoxazole and nysfungin). It was confirmed that the synthesized immunogen was highly antigenic and elicited specific antibody responses in BALB/C mice against EF.

  10. Antibody-based screening for hereditary nonpolyposis colorectal carcinoma compared with microsatellite analysis and sequencing

    DEFF Research Database (Denmark)

    Christensen, Mariann; Katballe, Niels; Wikman, Friedrik;


    BACKGROUND: Germline mutations in the DNA mismatch repair genes, MSH2, MLH1, and others are associated with hereditary nonpolyposis colorectal cancer (HNPCC). Due to the high costs of sequencing, cheaper screening methods are needed to identify HNPCC cases. Ideally, these methods should have a hi...

  11. Insight into screening immunoglobulin gene combinatorial libraries in a phage display vector: a tale of two antibodies. (United States)

    Kakinuma, A; Portolano, S; Chazenbalk, G; Rapoport, B; McLachlan, S M


    Combinatorial libraries of immunoglobulin genes in "phage display" vectors are a powerful tool for obtaining antigen-specific antibody fragments. To date, this approach has been used to isolate abundant, but not rare, human autoantibodies of IgG class. We have compared the relative efficiencies of panning pComb3 libraries made from intrathyroidal plasma cells for abundant human autoantibodies to thyroid peroxidase (TPO) and rare autoantibodies to the thyrotropin receptor (TSHR). TPO-specific Fab were readily obtained from a library using three different forms of recombinant antigen, (i) purified TPO, (ii) impure TPO in culture medium and, (iii) TPO expressed on the surface of CHO cells. In contrast, TSHR-specific Fab were not isolated. This was the case despite repeated pannings of six libraries from three optimal patients (IgG/kappa and IgG/lambda libraries for each patient). Both purified recombinant TSHR and CHO cells expressing TSHR on their surface were used. Library enrichment was observed on some screenings. However, Fab expressed by individual clones or from enriched libraries were not specific as determined by (i) binding to purified, radio-labeled antigen, (ii) FACS analysis of TSHR on intact CHO cells and, (iii) inhibition of radiolabeled TSH binding. Remarkably, in screening for both TPO- and TSHR-specific Fab, neither library enrichment nor the retention of cDNA inserts of the correct size correlated with obtaining Fab with the antigenic specificity sought. Indeed, excellent enrichment could be observed with conditioned medium from untransfected cells. Our data suggest that the key to isolating rare antibodies from phage display libraries is not the creation of vast libraries of greater diversity or even the development of more stable vectors. Rather, success in this endeavor appears to require reducing the "noise" of non-specific clones in a moderately sized library.

  12. Monoclonal neutralizing antibodies against EV71 screened from mice immunized with yeast-produced virus-like particles. (United States)

    Lin, Tao; Xianyu, Lingzhi; Lyu, Songya


    Periodic outbreaks of hand, foot and mouth disease (HFMD) occur in children under 5 years old, and can cause death in some cases. The C4 strain of enterovirus 71 (EV71) is the main pathogen that causes HFMD in China. Although no drugs against EV71 are available, some studies have shown that candidate vaccines or viral capsid proteins can produce anti-EV71 immunity. In this study, female BABL/c mice (6-8 weeks old) were immunized with virus-like particles (VLPs) of EV71 produced in yeast to screen for anti-EV71 antibodies. Two hybridomas that could produce neutralizing antibodies against EV71 were obtained. Both neutralizing mAbs (D4 and G12) were confirmed to bind the VP1 capsid protein of EV71, and could protect >95% cells from 100 TCID50 EV71 infection at 25 µg/mL solution (lowest concentration). Those two neutralizing mAbs identified in the study may be promising candidates in development for mAbs to treat EV71 infection, and utilized as suitable reagents for use in diagnostic tests and biological studies.

  13. Quality control and evaluation of human immunodeficiency virus antibody assays used for screening donated blood in China

    Institute of Scientific and Technical Information of China (English)



    During 2004, a total of 124 batches of HIV antibody ELISAs from domestic and overseas manufacturers, comprising approximately 60 million tests, were tested for quality and released for screening blood in China. The inter- and intra-batch variation, specificity, and sensitivity were evaluated using a laboratory panel and clinical samples. The inter-batch variation was less than 15% and only 2 of 12 assays had intra-batch variation of less than 20% for 4 dilutions of a control specimen.257 samples confirmed positive for HIV antibody and 4826 negative samples from different regions in China were used to evaluate the sensitivity and specificity of the assays. The results showed that the sensitivity is in the range from 93.7% to 100% for assays sampled directly from the manufacturers,and 91.4%-99.6% for those retrieved from the consumers; the specificity was in the range from 97.88% to 99.97%. The testing environment may vary in different regions of China. Therefore, manufacturers should provide robust assays to satisfy the requirements of these diverse environments, and especially reduce the intra-assay variation and improve the stability of the kits.

  14. Monoclonal neutralizing antibodies against EV71 screened from mice immunized with yeast-produced virus-like particles

    Institute of Scientific and Technical Information of China (English)

    Tao; Lin; Lingzhi; Xianyu; Songya; Lyu


    Periodic outbreaks of hand, foot and mouth disease(HFMD) occur in children under 5 years old, and can cause death in some cases. The C4 strain of enterovirus 71(EV71) is the main pathogen that causes HFMD in China. Although no drugs against EV71 are available, some studies have shown that candidate vaccines or viral capsid proteins can produce anti-EV71 immunity. In this study, female BABL/c mice(6–8 weeks old) were immunized with virus-like particles(VLPs) of EV71 produced in yeast to screen for anti-EV71 antibodies. Two hybridomas that could produce neutralizing antibodies against EV71 were obtained. Both neutralizing m Abs(D4 and G12) were confirmed to bind the VP1 capsid protein of EV71, and could protect > 95% cells from 100 TCID50 EV71 infection at 25 μg/m L solution(lowest concentration). Those two neutralizing m Abs identified in the study may be promising candidates in development for m Abs to treat EV71 infection, and utilized as suitable reagents for use in diagnostic tests and biological studies.

  15. Platelet antibody screening by flow cytometry is more sensitive than solid phase red cell adherence assay and lymphocytotoxicity technique: a comparative study in Thai patients. (United States)

    Buakaew, Jarin; Promwong, Charuporn


    The objective of this study was to compare the sensitivity and specificity of lymphocytotoxicity test (LCT), solid phase red cell adherence assay (SPRCA) and flow cytometry in detecting platelet reactive antibodies against human leukocyte antigens (HLA) class I and human platelet antigens (HPA). Sera from 38 thrombocytopenic patients and 5 mothers of thrombocytopenic newborns were screened for platelet reactive antibodies by these three methods using screening platelets and/or lymphocytes panels derived from six subjects. The sensitivity and specificity of each method and levels of agreement were analysed. HLA antibodies were found in 18, 17 and 19 out of 43 patients' sera tested by LCT, SPRCA and flow cytometry, respectively. Four out of 43 patients' sera were reactive against HPA by flow cytometry, but were reactive to only 2 sera by SPRCA. Using flow cytometry as the reference method, the sensitivities/specificities of SPRCA and LCT in HLA antibody detection were 84.21/95.83% and 94.73/100%, respectively, with a good strength of agreement. SPRCA had 50% sensitivity and 100% specificity in HPA antibody detection compare to flow cytometry. Flow cytometry appeared to be the most sensitive technique compared with SPRCA and LCT for both HPA and HLA antibody screening. SPRCA sensitivity was too low for HPA antibody detection, but this might be because of the small number of samples. There was one serum from the mother of a baby suffering neonatal alloimmune thrombocytopenia (NAIT), in whom SPRCA could not detect HPA antibodies, while flow cytometry came out positive. Therefore, SPRCA should not be used in NAIT investigation and flow cytometry should be employed instead.

  16. Label-free Fab and Fc affinity/avidity profiling of the antibody complex half-life for polyclonal and monoclonal efficacy screening. (United States)

    Read, Thomas; Olkhov, Rouslan V; Williamson, E Diane; Shaw, Andrew M


    A unified approach to affinity screening for Fab and Fc interactions of an antibody for its antigen and FcγR receptor has been developed. An antigen array is used for the Fab affinity and cross-reactivity screening and protein A/G proxy is the FcγR receptor. The affinities are derived using a simple 1:1 binding model with a consistent error analysis. The association and dissociation kinetics are measured over optimised times for accurate determination. The Fab/Fc affinities are derived for ten antibodies: mAb-actin (mouse), pAb-BSA (sheep), pAb-collagen V (rabbit), pAb-CRP (goat), mAb-F1 (mouse), mAbs (mouse) 7.3, 12.3, 29.3, 36.3 and 46.3 raised against LcrV in Yersinia pestis. The rate of the dissociation of antigen-antibody complexes relates directly to their immunological function as does the Fc-FcγR complex and a new half-life plot has been defined with a Fab/Fc half-life range of 17-470 min. The upper half-life value points to surface avidity. Two antibodies that are protective as an immunotherapy define a Fab half-life >250 min and an Fc half-life >50 min as characteristics of ideal interactions which can form the basis of an antibody screen for immunotherapy.

  17. A space-time analysis of Mycoplasma bovis: bulk tank milk antibody screening results from all Danish dairy herds in 2013-2014

    DEFF Research Database (Denmark)

    Arede, Margarida; Nielsen, Per Kantsø; Ahmed, Syed Sayeem Uddin;


    Mycoplasma bovis is an important pathogen causing severe disease outbreaks in cattle farms. Since 2011, there has been an apparent increase in M. bovis outbreaks among Danish dairy cattle herds. The dairy cattle industry performed cross-sectional antibody screening for M. bovis on four occasions,...

  18. Generation of high-affinity fully human anti-interleukin-8 antibodies from its cDNA by two-hybrid screening and affinity maturation in yeast. (United States)

    Ding, Ling; Azam, Mark; Lin, Yu-Huei; Sheridan, James; Wei, Shuanghong; Gupta, Gigi; Singh, Rakesh K; Pauling, Michelle H; Chu, Waihei; Tran, Antares; Yu, Nai-Xuan; Hu, Jiefeng; Wang, Wei; Long, Hao; Xiang, Dong; Zhu, Li; Hua, Shao-Bing


    We have developed a technology for rapidly generating novel and fully human antibodies by simply using the antigen DNA. A human single-chain variable fragment (scFv) antibody library was constructed in a yeast two-hybrid vector with high complexity. After cloning cDNA encoding the mature sequence of human interleukin-8 (hIL8) into the yeast two-hybrid system vector, we have screened the human scFv antibody library and obtained three distinct scFv clones that could specifically bind to hIL8. One clone was chosen for further improvement by a novel affinity maturation process using the error-prone PCR of the scFv sequence followed by additional rounds of yeast two-hybrid screening. The scFv antibodies of both primary and affinity-matured scFv clones were expressed in E. coli. All purified scFvs showed specific binding to hIL8 in reciprocal coimmunoprecipitation and ELISA assays. All scFvs, as well as a fully human IgG antibody converted from one of the scFv clones and expressed in the mammalian cells, were able to effectively inhibit hIL8 in neutrophil chemotaxis assays. The technology described can generate fully human antibodies with high efficiency and low cost.

  19. Pathogen-free screening of bacteria-specific hybridomas for selecting high-quality monoclonal antibodies against pathogen bacteria as illustrated for Legionella pneumophila. (United States)

    Féraudet-Tarisse, Cécile; Vaisanen-Tunkelrott, Marja-Liisa; Moreau, Karine; Lamourette, Patricia; Créminon, Christophe; Volland, Hervé


    Antibodies are potent biological tools increasingly used as detection, diagnostic and therapeutic reagents. Many technological advances have optimized and facilitated production and screening of monoclonal antibodies. We report here an original method to screen for antibodies targeting biosafety level 2 or 3 pathogens without the fastidious handling inherent to pathogen use. A double ELISA screening was performed using as coated antigen transformed Escherichia coli expressing at its surface a protein specific to the pathogenic bacteria versus control untransformed E. coli. This method was applied to Legionella, using the surface-exposed Mip protein (macrophage infectivity potentiator). This screening proved to be an excellent means of selecting mAbs that bind Legionella pneumophila 1 surface-exposed Mip protein. This method also appears more biologically relevant than screening using the recombinant Mip protein alone and less tedious than a test performed directly on Legionella bacteria. We obtained 21 mAbs that bind strongly to L. pneumophila serogroups 1 to 13, and we validated their use in a rapid ELISA (performed in 4.5 h) and an immunochromatographic test (20 min).

  20. Monoclonal antibody based immunoassays to screen for alpha-thalassemia in adults

    Energy Technology Data Exchange (ETDEWEB)

    Epstein, N.; Than, K.A. Culp, K.M. [Isolab, Inc., Akron, OH (United States)] [and others


    Alpha-thalassemia (alpha-thal) is characterized by the absence or reduction in synthesis of the alpha-globin chain due to either deletions or other abnormalities involving the alpha-globin genes located on the short arm of chromosome 16. The diploid cells have four alpha chain genes. The deletion of one, two, three or all four of these genes could result in mild to a complete alpha chain deficiency known as the Hydrops fetalis syndrome or alpha-thal-1, which causes fetal death. It is important to develop a sensitive test to detect carriers of alpha-thal-1 trait for genetic counseling. It has recently been observed that the presence of minute amounts of zeta-globin chains (0.01-1%) could serve as a biological marker of alpha-thal carriers. Because high sensitivity is required, we constructed a monoclonal antibody-based immunoassay which can be analyzed either by colorimetric or fluorimetric methods. By testing blood samples from individuals of Southeast Asian ancestry, we were able to show that various forms and combinations of deletions or inactivations of two or three alpha-globin genes results in alpha-thalassemia conditions that have elevated levels of the zeta-chain. Sensitivity achieved in these tests was < 0.1% zeta chain, or as low as 5 ng zeta-chain. Data correlate with results from reversed phase HPLC.

  1. Multipin peptide libraries for antibody and receptor epitope screening and characterization. (United States)

    Tribbick, Gordon


    It has been nearly 15 years since the papers describing the fully systematic epitope mapping approach both for the so-called "continuous" epitopes [Proc. Natl. Acad. Sci. U. S. A. 81 (1984) 3998] and "discontinuous" epitopes [Mol. Immunol. 23 (1986) 709] were published. These seminal papers laid the conceptual foundation for all subsequent developments where a combinatorial approach is applied. Dr. Mario Geysen, the 2000 Kilby Laureate, can certainly lay claim to be the "father of combinatorial chemistry" ( In this review, I will focus on the aspects of the Multipin technology as they apply to antibody and receptor epitope mapping. Much of what will be presented applies equally well to other applications where peptide libraries (PepSets) and combinatorial approaches are used [Rodda, S.J., 1996. T-cell epitope mapping with synthetic peptides and peripheral blood mononuclear cells. In: Morris, G.E. (Eds.), Methods in Molecular Biology, Vol. 66: Epitope Mapping Protocols. Humana Press, Totowa, NJ, Chap. 30, p. 363; Int. J. Pept. Protein Res. 42 (1993) 384; J. Biol. Chem. 271 (1996) 5603]. Factors and techniques that influence the use of the Multipin method for successful epitope mapping will be presented.

  2. 献血人群红细胞血型意外抗体的筛选%Screening unexpected antibody of red cells blood group in the blood donors

    Institute of Scientific and Technical Information of China (English)

    祝宏; 洪小珍; 吴亚玲; 励晓涛; 马开荣; 兰小飞; 朱发明


    目的:分析献血人群中红细胞血型意外抗体的情况,并对抗体特性进行确认.方法:应用O型红细胞结合PK7200血型检测系统筛选红细胞血型意外抗体,利用谱细胞鉴定抗体特性.结果:在献血人群中红细胞血型意外抗体阳性率为0.025%,意外抗体以抗M和冷凝集素为主.发现两例类孟买型血型和两例p血型.结论:献血人群中存在低比例的红细胞血型意外抗体,在献血者血型检测中应加以重视.%Objective;To determine unexpected antibody of red cells blood group in the blood donors and identify the characteristics of the antibody. Methods: The unexpected antibody was screened by 0 blood group cells and PK7200 system. Characteristics of the antibody were identified by panel cells.. Results: The prevalence rate of unexpected antibody was 0.025% in the blood donors. The common unexpected antibodies were anti - M and cold ag-glutinin. Two cases with para - Bombay phenotype and two cases with p phenotype were found. Conclusion; Low prevalence of unexpected antibody existed in the blood donors. It is important to screen unexpected antibody in donor's blood grouping.

  3. Antibody binding site mapping of SARS-CoV spike protein receptor-binding domain by a combination of yeast surface display and phage peptide library screening. (United States)

    Zhang, Xiaoping; Wang, Jingxue; Wen, Kun; Mou, Zhirong; Zou, Liyun; Che, Xiaoyan; Ni, Bing; Wu, Yuzhang


    The receptor-binding domain (RBD) of severe acute respiratory syndrome coronavirus (SARS-CoV) spike (S) protein plays an important role in viral infection, and is a potential major neutralizing determinant. In this study, three hybridoma cell lines secreting specific monoclonal antibodies against the RBD of the S protein were generated and their exact binding sites were identified. Using yeast surface display, the binding sites of these antibodies were defined to two linear regions on the RBD: S(337-360) and S(380-399). Using these monoclonal antibodies in phage peptide library screening identified 10 distinct mimotopes 12 amino acids in length. Sequence comparison between native epitopes and these mimotopes further confirmed the binding sites, and revealed key amino acid residues involved in antibody binding. None of these antibodies could neutralize the murine leukemia virus pseudotyped expressing the SARS-CoV spike protein (MLV/SARS-CoV). However, these mAbs could be useful in the diagnosis of SARS-CoV due to their exclusive reactivity with SARS-CoV. Furthermore, this study established a feasible platform for epitope mapping. Yeast surface display combined with phage peptide library screening provides a convenient strategy for the identification of epitope peptides from certain antigenic proteins.

  4. Frequencies and Specificities of “Enzyme-Only” Detected Erythrocyte Alloantibodies in Patients Hospitalized in Austria: Is an Enzyme Test Required for Routine Red Blood Cell Antibody Screening?

    Directory of Open Access Journals (Sweden)

    Dietmar Enko


    Full Text Available The aim of this study was to determine the frequencies and specificities of “enzyme-only” detected red blood cell (RBC alloantibodies in the routine antibody screening and antibody identification in patients hospitalized in Austria. Routine blood samples of 2420 patients were investigated. The antibody screening was performed with a 3-cell panel in the low-ionic strength saline- (LISS- indirect antiglobulin test (IAT and with an enzyme-pretreated (papain 3-cell panel fully automated on the ORTHO AutoVue Innova System. The antibody identification was carried out manually with an 11-cell panel in the LISS-IAT and with an enzyme-pretreated (papain 11-cell panel. In total 4.05% (n=98 of all patients (n=2420 had a positive RBC antibody screening result. Of them 25.51% (25/98 showed “enzyme-only” detected specific or nonspecific RBC alloantibodies. Rhesus and Lewis system antibodies were found the only specificities of “enzyme-only” RBC alloantibodies: all in all 4.8% (4/98 were detected with anti-E, 3.06% (3/98 with anti-Lea, 3.06% (3/98 with anti-D after anti-D prophylaxis and 1.02% (1/98 with anti-e. In total, 14.29% (14/98 showed a nonspecific RBC alloantibody result with the enzyme test. The results of the present study demonstrate that a high number of unwanted positive reactions with the enzyme technique overshadows the detection of “enzyme-only” RBC alloantibodies. (Trial Registration: K-37-13.

  5. Identification of midgut microvillar proteins from Tenebrio molitor and Spodoptera frugiperda by cDNA library screenings with antibodies. (United States)

    Ferreira, A H P; Cristofoletti, P T; Lorenzini, D M; Guerra, L O; Paiva, P B; Briones, M R S; Terra, W R; Ferreira, C


    The objective of this study was to identify midgut microvillar proteins in insects appearing earlier (Coleoptera) and later (Lepidoptera) in evolution. For this, cytoskeleton-free midgut microvillar membrane from Spodoptera frugiperda (Lepidoptera) and Tenebrio molitor (Coleoptera) were used to raise antibodies. These were used for screening midgut cDNA expression libraries. Positive clones were sequenced, assembled and searched for similarities with gene/protein databases. The predicted midgut microvillar proteins from T. molitor were: cockroach allergens (unknown function), peritrophins (peritrophic membrane proteins), digestive enzymes (aminopeptidase, alpha-mannosidase) and unknown proteins. Predicted S. frugiperda midgut proteins may be grouped into six classes: (a) proteins involved in protection of midgut (thioredoxin peroxidase, aldehyde dehydrogenase, serpin and juvenile hormone epoxide hydrolase); (b) digestive enzymes (astacin, transporter-like amylase, aminopeptidase, and carboxypeptidase); (c) peritrophins; (d) proteins associated with microapocrine secretion (gelsolin, annexin); (e) membrane-tightly bound-cytoskeleton proteins (fimbrin, calmodulin) and (f) unidentified proteins. The novel approach is compared with others and microvillar function is discussed in the light of the predicted proteins.

  6. Impact analysis of autoantibody level and NR2 antibody level in neuropsychiatric SLE treated by methylprednisolone combined with MTX and DXM intrathecal injection. (United States)

    Wang, Jingyuan; Zhao, Yinhuan; Zhang, Jihui; Lei, Hongwei; Zhu, Guiqi; Fu, Bingbing


    The objective is to explore the clinical curative effects of methylprednisolone combined with MTX and DXM intrathecal injection in treating neuropsychiatric systemic lupus erythematosus (NPSLE) and its effects on autoantibody level and anti-N-methyl-D-aspartate receptor subtype NR2a/2b antibody (anti-NR2 antibody) level. Thirty six admitted NPSLE patients were treated by methylprednisolone combined with MTX and DXM intrathecal injection. Thirty six SLE patients without neuropsychiatric symptoms were selected as non-NPSLE group. Clinical indexes including SLE activity index, erythrocyte sedimentation rate (ESR), cerebrospinal fluid pressure (CSFP), cerebrospinal fluid protein were observed before and after treatment. Autoantibodies including anti-nuclear antibody (ANA), anti-double stranded DNA antibody (anti-dsDNA antibody), anti-extractable nuclear antigen antibody (ENA-Ab) were detected before and after treatment. Enzyme linked immunosorbent assay was used to detect NR2 antibody level before and after treatment in two groups. Upon treatment of methylprednisolone combined with MTX and DXM intrathecal injection, SLE activity index, ESR, CSFP, cerebrospinal fluid protein of 36 NPSLE patients were significantly decreased. Before treatment, positive rates of ANA, anti-dsDNA antibody, and anti-ENA antibody in both NPSLE group and non-NPSLE group had no significant difference. However, positive rate of anti-NR2 antibody in NPSLE group was significantly higher than that of non-NPSLE group. After treatment, positive rates of autoantibodies and anti-NR2 antibody in both NPSLE and non-NPSLE group were significantly decreased. Anti-NR2 antibody can be a screening index of NPSLE, and methylprednisolone combined with MTX and DXM intrathecal injection has significant curative effects and can effectively decrease autoantibody level and anti-NR2 antibody level.

  7. Antithyroglobulin antibody (United States)

    Thyroglobulin antibody; Thyroiditis - thyroglobulin antibody; Hypothyroidism - thyroglobulin antibody; Thyroiditis - thyroglobulin antibody; Graves disease - thyroglobulin antibody; Underactive thyroid - thyroglobulin antibody

  8. Confirmation of positive antibody screens by solid-phase red cell adherence assay using a tube technique method with polyethylene glycol enhancement. (United States)

    Gammon, R R; Lake, M; Velasquez, N; Prichard, A


    Our blood bank routinely screens donors for antibodies using a solid-phase red cell adherence (SPRCA) assay. Positive results are then confirmed using a tube technique with polyethylene glycol (PEG) enhancement due to reported higher specificity than with SPRCA. Over a 5-month period, 49,084 donor serum or plasma samples were tested using the SPRCA assay. Further identification of positive samples was performed using a PEG enhancement method. Testing was performed with strict adherence to the manufacturers' inserts. Of 49,084 samples, 313 (0.64%) were positive by the SPRCA assay. Of these, 99 (31.6%) samples remained positive when tested with PEG enhancement. The remaining 214 (68.4%) were negative, giving specificity for the SPRCA assay of 99.6 percent (48,985/ 49,199). We report a high specificity for antibody screening using the SPRCA assay. However, it is cost effective to perform a confirmatory tube test with PEG enhancement because 214 SPRCA assay samples were interpreted as having a negative antibody screen, thus allowing the release of valuable blood components for transfusion.

  9. Specificity of monoclonal anti-nucleosome auto-antibodies derived from lupus mice

    NARCIS (Netherlands)

    Kramers, K; Stemmer, C; Monestier, M; vanBruggen, MCJ; RijkeSchilder, TPM; Hylkema, MN; Smeenk, RJT; Muller, S; Berden, JHM


    Recently, anti-nucleosome antibodies, which do not bind to DNA or to individual histones, have been identified in longitudinal studies in lupus mice. These anti-nucleosome antibodies occur early in spontaneous SLE and are formed prior to other anti-nuclear specificities. However, nucleosomal epitope

  10. Screening for genes involved in antibody response to sheep red blood cells in the chicken, Gallus gallus. (United States)

    Geng, Tuoyu; Guan, Xiaojing; Smith, Edward J


    Antibody response, an important trait in both agriculture and biomedicine, plays a part in protecting animals from infection. Dissecting molecular basis of antibody response may improve artificial selection for natural disease resistance in livestock and poultry. A number of genetic markers associated with antibody response have been identified in the chicken and mouse by linkage-based association studies, which only define genomic regions by genetic markers but do not pinpoint genes for antibody response. In contrast, global expression profiling has been applied to define the molecular bases of a variety of biological traits through identification of differentially expressed genes (DEGs). Here, we employed Affimetrix GeneChip Chicken Genome Arrays to identify differentially expressed genes for antibody response to sheep red blood cells (SRBC) using chickens challenged with and without SRBC or chickens with high and low anti-SRBC titers. The DEGs include those with known (i.e., MHC class I and IgH genes) or unknown function in antibody response. Classification test of these genes suggested that the response of the chicken to intravenous injection of SRBC involved multiple biological processes, including response to stress or other different stimuli, sugar, carbohydrate or protein binding, and cell or soluble fraction, in addition to antibody response. This preliminary study thus provides an insight into molecular basis of antibody response to SRBC in the chicken.

  11. Construction of human phage antibody library and screening for human monoclonal antibodies of amylin%人源性噬菌体抗体库的构建及抗amylin抗体的初步筛选

    Institute of Scientific and Technical Information of China (English)

    公倩; 李常颖; 畅继武; 朱铁虹


    AIM- To screen monoclonal antibodies to amyiin from a constructed human phage antibody library and identify their antigenic specificity and combining activities. METHODS: The heavy chain Fd fragment and light chain of human immunoglobulin genes were amplified from peripheral blood lymphocytes of healthy donors using RT-PCR, and then inserted into phagemid pComb3XSS to generate a human phage antibody library. The insertion of light chain or heavy chain Fd genes were identified by PCR after the digestion of Sac I, Xba I , Xho I and Spe I. One of positive clones was analyzed by DNA sequencing. The specific anti-amylin clones were screened from antibody library against human amylin antigens and then the positive donas were determined by Phage-ELISA analysis. RESULTS; A Fab phage antibody library with 0.8x 108 members was constructed with the efficacy of about 70%. DNA sequence analysis indicated VH gene belonged to VH3 gene family and Vλ gene belonged to the Vλ gene family. Using human amylin as panning antigen, specific anti-amylin Fab antibodies were enriched by screening the library for three times. Phage-ELISA assay showed the positive clones had very good specificity to amylin antigen. CONCLUSION; The successful construction of a phage antibody library and the identification of anti-amylin Fab antibodies provide a basis for further study and preparation of human anti-amylin antibodies.%目的:构建人源性噬菌体抗体库,从中筛选胰淀素(amylin)单克隆抗体(mAb),测定其特异性及抗原结合活性.方法:从正常健康人的外周血淋巴细胞中提取总RNA,用RT-PcR方法扩增人免疫球蛋白Fd段和轻链基因,构建噬菌体抗体库.酶切和PcR鉴定后,阳性克隆进行DNA测序分析.用人amylin抗原对抗体库进行筛选富集,将得到的阳性克隆进行Phage-ELISA鉴定,结果进行统计学分析.结果:最终构建的抗体库库容约为0.8×108,酶切鉴定显示有插入片段,抗体库重组率为70%.阳性克

  12. Establishment of intein-mediated protein ligation under denaturing conditions: C-terminal labeling of a single-chain antibody for biochip screening. (United States)

    Sydor, Jens R; Mariano, Maria; Sideris, Steve; Nock, Steffen


    Intein-mediated protein ligation is a recently developed method that enables the C-terminal labeling of proteins. This technique requires a correctly folded intein mutant that is fused to the C-terminus of a target protein to create a thioester, which allows the ligation of a peptide with an N-terminal cysteine (1, 2). Here we describe the establishment of this method for the labeling, under denaturing conditions, of target proteins that are expressed insolubly as intein fusion proteins. A GFPuv fusion protein with the Mycobacterium xenopi gyrA intein was expressed in inclusion bodies in Escherichia coli and initially used as a model protein to verify intein cleavage activity under different refolding conditions. The intein showed activity after refolding in nondenaturing and slightly denaturing conditions. A construct of the same intein with an anti-neutravidin single-chain antibody was also expressed in an insoluble form. The intein-mediated ligation was established for this single chain antibody-intein fusion protein under denaturing conditions in 4 M urea to prevent significant precipitation of the fusion protein during the first refolding step. Under optimized conditions, the single-chain antibody was labeled with a fluorescent peptide and used for antigen screening on a biochip after final refolding. This screening procedure allowed the determination of binding characteristics of the scFv for avidin proteins in a miniaturized format.

  13. Computational screening of the human TF-glycome provides a structural definition for the specificity of anti-tumor antibody JAA-F11.

    Directory of Open Access Journals (Sweden)

    Matthew B Tessier

    Full Text Available Recombinant antibodies are of profound clinical significance; yet, anti-carbohydrate antibodies are prone to undesirable cross-reactivity with structurally related-glycans. Here we introduce a new technology called Computational Carbohydrate Grafting (CCG, which enables a virtual library of glycans to be assessed for protein binding specificity, and employ it to define the scope and structural origin of the binding specificity of antibody JAA-F11 for glycans containing the Thomsen-Friedenreich (TF human tumor antigen. A virtual library of the entire human glycome (GLibrary-3D was constructed, from which 1,182 TF-containing human glycans were identified and assessed for their ability to fit into the antibody combining site. The glycans were categorized into putative binders, or non-binders, on the basis of steric clashes with the antibody surface. The analysis employed a structure of the immune complex, generated by docking the TF-disaccharide (Galβ1-3GalNAcα into a crystal structure of the JAA-F11 antigen binding fragment, which was shown to be consistent with saturation transfer difference (STD NMR data. The specificities predicted by CCG were fully consistent with data from experimental glycan array screening, and confirmed that the antibody is selective for the TF-antigen and certain extended core-2 type mucins. Additionally, the CCG analysis identified a limited number of related putative binding motifs, and provided a structural basis for interpreting the specificity. CCG can be utilized to facilitate clinical applications through the determination of the three-dimensional interaction of glycans with proteins, thus augmenting drug and vaccine development techniques that seek to optimize the specificity and affinity of neutralizing proteins, which target glycans associated with diseases including cancer and HIV.

  14. Computational screening of the human TF-glycome provides a structural definition for the specificity of anti-tumor antibody JAA-F11. (United States)

    Tessier, Matthew B; Grant, Oliver C; Heimburg-Molinaro, Jamie; Smith, David; Jadey, Snehal; Gulick, Andrew M; Glushka, John; Deutscher, Susan L; Rittenhouse-Olson, Kate; Woods, Robert J


    Recombinant antibodies are of profound clinical significance; yet, anti-carbohydrate antibodies are prone to undesirable cross-reactivity with structurally related-glycans. Here we introduce a new technology called Computational Carbohydrate Grafting (CCG), which enables a virtual library of glycans to be assessed for protein binding specificity, and employ it to define the scope and structural origin of the binding specificity of antibody JAA-F11 for glycans containing the Thomsen-Friedenreich (TF) human tumor antigen. A virtual library of the entire human glycome (GLibrary-3D) was constructed, from which 1,182 TF-containing human glycans were identified and assessed for their ability to fit into the antibody combining site. The glycans were categorized into putative binders, or non-binders, on the basis of steric clashes with the antibody surface. The analysis employed a structure of the immune complex, generated by docking the TF-disaccharide (Galβ1-3GalNAcα) into a crystal structure of the JAA-F11 antigen binding fragment, which was shown to be consistent with saturation transfer difference (STD) NMR data. The specificities predicted by CCG were fully consistent with data from experimental glycan array screening, and confirmed that the antibody is selective for the TF-antigen and certain extended core-2 type mucins. Additionally, the CCG analysis identified a limited number of related putative binding motifs, and provided a structural basis for interpreting the specificity. CCG can be utilized to facilitate clinical applications through the determination of the three-dimensional interaction of glycans with proteins, thus augmenting drug and vaccine development techniques that seek to optimize the specificity and affinity of neutralizing proteins, which target glycans associated with diseases including cancer and HIV.

  15. Screening of scFvs against cTnI from Phage Display Antibody Library and Their Expression in E.coli Rosetta

    Institute of Scientific and Technical Information of China (English)

    WEI Jing-yan; ZHANG Han-qi; JIN Qin-han; LI Wei; LUO Gui-min; LI Shan-yu; MU Ying; ZHU Xue-jun; LIU Lei; GAO Li-zeng; SONG Da-qian; SUN Zhi-wei; YAN Gang-lin


    The single chain variable fragments of antibodies(scFvs) against eTnI were screened from the phage display antibody library by using cTnI as the target antigen. After four rounds of panning, four clones(H2, G5,A9, B9) from the phage display antibody library were verified to show higher binding affinity for cTnI by ELISA and to contain the variable region genes of the light and heavy chains of scFvs by sequencing. The variable region genes of scFvs H2 and G5 were successfully amplified by polymerase chain reactions (PCR)and cloned into expression vector pPELB and expressed as a soluble protein in E. coli Rosetta, whose expression yield was about 2% of total proteins. The expressed proteins were purified by nickel(Ni) affinity chromatography and a single band is shown in the position of 28 kDa on SDS-PAGE. The western blot analysis result verifies that the expressed scFv proteins are capable of binding with monoclonal antibodies against hexa-histidine, indicating that they are hexa-histidin-tagged aim proteins. The immunoassay demonstrates that the expressed scFv proteins are able to specifically react with cTnI molecules. The association constant (KA)values range from 1.2×104 to 1.7×105 L/mol that are correspondent to the affinities of polyclonal antibodies against cTnI from rabbits. These antibodies can be valuable reagents for the immunoassay of cTnI.

  16. Screening and identification of neutralizated single-chain antibody of anti-glomerular basement membrane antibody%中和抗肾小球基底膜抗体的单链抗体的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    肖静; 刘章锁; 王沛; 黄留玉; 宋宏彬; 赵明辉


    Objective To screen a human single-chain variable fragments(scFv)against antiGBM antibody.Methods Using phage display technique,the phage antibody library was panned by antiglomerular basement membrane(GBM)antibody which was coated in a micro-titer plate,one clone was found to have high affinity to anti-GBM antibody.The DNA sequence of the positive clone was determined.Results Along with the increase of rounds anti-GBM antibody specific phage antibody was highly enriched and screening efficiency was increased 137 folds than the firest round.ELISA and competition inhibition assay showed that the scFv had a specific combination character with anti-GBM antibody.DNA sequencing confirmed that the whole gene of scFv was 750 bp,and in accordance with humanized single-chain variable region antibody sequence structure.Conclusion The results suggested that the scFv fragment to anti-GBM antibody could be successfully selected by recombinant phage antibody technique,which will laid an experimental foundation for further research of the therapy of Goodpasture syndrome.%目的 制备人抗肾小球基底膜(GBM)抗体的特异性人源化单链可变区抗体.方法 采用噬菌体表面展示技术,获得一个与人抗GBM抗体结合活性较强的单链可变区抗体片段的阳性克隆,并对该克隆进行DNA序列测定分析.结果 对噬菌体单链可变区抗体库经过3轮筛选后,与第1轮相比富集了137倍.噬菌体抗体与人抗GBM抗体的结合活性其中有35株克隆ELISA的吸光度较高.对这些噬菌体抗体进行交叉反应后,确定其中有10株交叉反应较弱.确定1株(C31)阳性克隆提取质粒,进行DNA序列测定,大小为750 bp,并符合人源化单链可变区抗体的序列结构.结论 应用噬菌体展示技术成功获得人-抗GBM抗体的单链可变区抗体基因,为临床上治疗Goodpasture综合征奠定实验基础.

  17. Available means: manifestations of Aristotle's three modes of rhetorical appeal in antinuclear fiction

    Energy Technology Data Exchange (ETDEWEB)

    Mannix, P.J.


    The abundance of sympathetic scientists, military men and clergymen in antinuclear fiction reflects a public perception that authorities speak most knowledgeably about an issue. Other antinuclear works employ characters with less traditional ethical appeals: nurturing women, vital youths, and even infallible computers. Antinuclear fiction uses enthymeme and example to reflect the history of the nuclear weapons debate. Some works attach the immorality of the weapons by examining the moral dilemmas of nuclear scientists. Others admit the permanence of the nuclear threat. By arousing emotions, fiction is capable of mobilizing its audience's active support for the ideas it presents. The principal emotions that various antinuclear works arouse highlight the close relationship between literature and rhetoric. The most dominant emotions, pity and fear, are the two Aristotle links to tragedy. Scorn, the principal emotion that Dr. Strangelove arouses - is the crucial emotion on which all satire depends. However, the other principal emotion in anti-nuclear fiction - hope - has principally a rhetorical function ensuring that the feelings the works provoke will be channeled constructively.

  18. HIV antibody preliminary screening in 113 229 inpatients%113229例住院患者 HIV 抗体初筛结果分析

    Institute of Scientific and Technical Information of China (English)

    王梅芬; 王庆芳; 刘家田; 李建丽; 王万海


    目的:了解某大型医院住院患者人类免疫缺陷病毒( HIV)抗体初筛阳性率及其在临床科室中分布情况。方法对113229例住院患者进行HIV抗体初筛检测,比较不同性别患者的阳性率,分析HIV抗体阳性患者在不同科室的分布情况。结果 HIV抗体初筛阳性152例,阳性率为0.134%;其中男91例(0.161%),女61例(0.107%),不同性别间阳性率比较差异有统计学意义(P<0.05)。 HIV抗体初筛阳性患者分布于20个科室,其中高发科室有耳鼻喉科、骨科、妇科、泌尿外科、普外科、神经内科、皮肤科、肿瘤科、眼科。这些科室的HIV抗体初筛阳性者占总筛查阳性的67.1%。结论住院患者中存在较高的HIV抗体初筛阳性率,且科室分布广泛。%Objective To investigate the positive rate of HIV antibody preliminary screening and the distribution of the clinical departments in general hospital inpatients.Methods HIV antibody preliminary screening detection was conducted in 113 229 cases of patients.The positive rate between different genders was compared , and the distribution of patients with positive HIV antibody in different departments was analyzed.Results Totally 152 plasma samples were positive by the antibody screening test for HIV, and the positive rate was 0.134%, including 91 males and 61 females (P<0.05).The 152 positive cases covered 20 clinical departments and the major clinical departments included otolaryngological department, orthopaedics department, gy-naecology department, urology department, surgery department, neurology department, dermatological department, oncology de-partment and ophthalmology department.The patients with positive HIV antibody undergoing preliminary screening of these de-partments accounted for 67.1%of the total screening.Conclusion The positive rate of HIV antibody screening among inpatients in our hospital is high with a wide range of department

  19. Screening for antibodies against Aleutian disease virus (ADV) in mink. Elucidation of dubious results by additive counterimmunoelectrophoresis

    DEFF Research Database (Denmark)

    Uttenthal, Åse


    In order to distinguish true positive results in counterimmunoelectrophoresis from false positive ones an additive counterimmunoelectrophoresis was developed. The method was tested on selected mink serum samples as part of a routine testing for antibodies towards Aleutian disease virus on 3 million...... blood samples. The procedure of the method is, that a known positive serum sample is mixed with the patient serum to be tested. The result from a false positive sample will be one precipitin line towards virus and one nonspecific line. If the serum sample is a true positive one, the antibodies...... originating from the patient serum will be added to the antibodies in the standard positive serum giving only one precipitin line. The system is further extended by testing the serum samples towards an antigen preparation containing all the cellular components but free from virus....

  20. Thyroid Antibodies, Autoimmunity and Cognitive Decline: Is There a Population-Based Link

    Directory of Open Access Journals (Sweden)

    Kate Napthali


    Full Text Available Background: Autoimmunity is considered an uncommon but under-recognised cause of cognitive decline. Methods: Serum samples from 3,253 randomly selected subjects enrolled in the Hunter Community Study, aged 55-85 years, were assayed for thyrotropin stimulatory hormone, anti-thyroid peroxidase antibodies (TPO-Ab, anti-nuclear antibodies (ANA and extractable nuclear antigens (ENA. Cognitive function was assessed using the Audio Recorded Cognitive Screen (ARCS tool. Results: TPO-Ab were found in 8.4% and ANA in 27.9% of the study population, of whom 3% had positive ENA findings. No relationship was found between the ARCS score and either TPO-Ab (coefficient = 0.133; 95% CI -0.20, 0.82, p = 0.616, ANA at a low (coefficient = 1.01; 95% CI -2.58, 0.55, p = 0.203 or a high titre (coefficient = -0.65; 95% CI -2.59, 1.28, p = 0.508, or ENA antibodies (coefficient = 5.12; 95% CI -0.53, 10.77; p = 0.076. Conclusions: Autoantibody findings are common in an aging population and are not associated with cognitive decline.

  1. Evaluation of a blocking ELISA for screening of antibodies against porcine reproductive and respiratory syndrome (PRRS) virus

    DEFF Research Database (Denmark)

    Sørensen, K.J.; Bøtner, Anette; Madsen, E.S.


    A blocking Elisa was developed for the detection of antibodies against PRRS virus with a view to satisfying the need for examination of blood samples on a large scale. The test was evaluated in comparison with an indirect Elisa and the immunoperoxidase monolayer assay. The blocking Elisa was sens......A blocking Elisa was developed for the detection of antibodies against PRRS virus with a view to satisfying the need for examination of blood samples on a large scale. The test was evaluated in comparison with an indirect Elisa and the immunoperoxidase monolayer assay. The blocking Elisa...

  2. Evaluation of Anti-Nuclear antibody test results in clinical practice


    Nevreste Çelikbilek; Birsen Özdem; Ziya Cibali Açıkgöz


    Amaç: Bu çalışmanın amacı 2009-2011 yıllarında laboratuvarımızda yapılan ANA (antinükleer antikor) tetkik sonuçlarının retrospektif olarak değerlendirilmesidir. Yöntemler: Bu dönemde laboratuvarımızda toplam 5068 serum örneğinde indirekt immünofloresans antikor yöntemiyle (IIFA) ANA varlığı araştırıldı. Rastgele örnekleme ile seçilen ANA-pozitif 982 olgu cinsiyet dağılımı, ışıma düzeyleri ve paternleri, dsDNA sonuçları ve ekstrakte edilebilir nükleer antijen (ENA) profilleri açısından incelen...

  3. Antinuclear antibody levels in Polymorphic Light Eruption and their relation to the severity of disease

    Directory of Open Access Journals (Sweden)

    Iffat Hassan


    Full Text Available Introduction: PLE is an idiopathic photodermatosis characterised by a polymorphic eruption ranging from papulo-vesicular lesions to large plaques, located predominantly in a photoexposed distribution. It is an acquired disease and is the most common idiopathic photodermatosis. It is characterised by recurrent abnormal delayed reaction to sunlight. PLE is the most common idiopathic photodermatosis, the prevalence of which has been estimated to be around 10-20% in USA, England and Ireland. Previous studies have shown elevated levels of ANA in 2.9-19% of patients with PLE. Aim: The aim of our study was to determine the frequency of ANA positivity in a cohort of patients of ethnic kashmiri origin with Polymorphic light eruption and to examine whether there is any relation of their levels with the severity of disease.Methods: Patients with Polymorphic light eruption clinically who attended the Outpatient Deptt. of Dermatology GMC Srinagar were referred to the Deptt. of Biochemistry GMC Srinagar where patients blood samples were analysed for ANA index by ELISA method (BRIO SEAC ITALY.Results: Our study consisted of 36 patients (with 23 males and 13 females with age group ranging between 15-65 years presenting with typical clinical features of PLE without associated autoimmune connected tissue diseases like discoid lupus erythematosus or systemic lupus erythematosus and 20 healthy age and sex matched controls. Two patients (1 male and 1female showed positive results and 1 patient (female showed equivocal results. Among the control group one patient showed ANA positivity. Thus total frequency of ANA positivity of of 5.55% was observed among the cases and 5% among the controls with frequency of 4.34% in males and 7.69% in females.Conclusion: ANA levels were found in 5.55% of patients with PLE, however there is no relation between the levels of ANA in PLE and with the severity of disease (p value >0.05.

  4. 2002~2011年某院HIV抗体筛查结果分析%Analysis of HIV antibody screening results in a hospital during 2002-2011

    Institute of Scientific and Technical Information of China (English)

    吴红; 罗显华; 罗丽; 张更建; 谌宏运; 李进芹; 陈龙庆; 张信江


    目的:分析遵义医学院附属医院人类免疫缺陷病毒(HIV)抗体筛查结果,侧面了解遵义地区艾滋病(AIDS)流行状况,为AIDS的防治提供依据。方法选取遵义医学院附属医院2002年1月至2011年12月需要输血、外科手术的住院患者及性病门诊的可疑感染HIV患者共201052例为研究对象,采用酶联免疫吸附法(ELLSA法)进行HIV抗体初筛,初筛阳性者送贵州省或遵义市疾控中心 HIV 确诊实验室行蛋白质印迹试验(WB)确诊。结果2002~2011年10年间该院共检测出HIV抗体阳性者355例,其中呼吸科最多,共54例(15.21%);年龄2 d至78岁;涉及职业广泛,以自由职业与无固定职业者居多(49.86%)。结论对需要输血、外科手术、围产保健及性病门诊可疑感染HIV的患者进行HIV抗体检测是发现HIV/AIDS的重要途径,加强对上述有关人群进行HIV抗体筛查在及早发现HIV/AIDS、避免HIV的医源性传播方面具有重要意义。%Objective To analyze the screening results of human immunodeficiency virus (HIV) antibody for understand-ing the prevalence situation of acquired immune deficiency syndrome (AIDS) in Zunyi area from one aspect and providing the ba-sis for AIDS prevention. Methods A total of 201 052 inpatients with blood transfusion and surgical operation and patients with suspected HIV infection in the venereal disease outpatient department of Affiliated Hospital of Zunyi Medical College from Jan-uary 2002 to December 2011 were selected as the research subjects. HIV antibody was preliminarily screened by adopting ELISA. Then the cases of positive screening were performed the Western blotting for confirmation in the center of disease control of Zunyi City or Guizhou Province. Results Totally 355 cases of HIV antibody positive were detected out during these 10 years of 2002-2011,among them majority were from the respiratory department(54 cases,15.21%);age was from 2 d to 78

  5. Investigation of Antiphosphatidyl-Serine Antibody and Antiphosphatidyl-Inositol Antibody in Ischemic Stroke Patients

    Directory of Open Access Journals (Sweden)

    Hirohisa Okuma


    Full Text Available Antiphospholipid syndrome is characterized by arterial or venous thrombosis and the presence of antiphospholipid antibodies (aPL. We measured β2-GPI aCL, IgGaCL, LA, antiphosphatidyl-serine antibody (PS, and antiphosphatidyl-inositol antibody (PI in each patient at one month after the onset of stroke. In addition, carotid artery echography was performed in patients positive for PI or PS. Among the 250 patients, 13.6% (34/250 were positive for either PI or PS, and 6.8% (17/250 were positive for both. Carotid artery echography performed on these 34 patients showed that the frequencies of increased intimal-medial thickness (IMT of 1.1 mm or more, plaque, and carotid artery stenosis of 50% or more were all significantly higher in patients positive for antinuclear antibody than those negative for the antibody (P<.05. PI and PS are associated with antinuclear antibody and precipitation of atherosclerosis. Ischemic stroke patients with SLE frequently showed a variety of antiphospholipid-protein antibodies.

  6. Identification of the specificity of isolated phage display single-chain antibodies using yeast two-hybrid screens

    DEFF Research Database (Denmark)

    Rasmussen, Nicolaj; Ditzel, Henrik


    A method is described for the identification of the antigen recognised by an scFv isolated from an antibody phage display library using selection against a complex mixture of proteins (e.g. intact cells, purified cell surface membranes, and tissue sections). The method takes advantage of a yeast...... two-hybrid system that additionally allows for reorganization of post-translational modifications to the bait and target proteins. This technique is therefore especially useful for identifying surface-expressed antigens....

  7. Construction of normal human IgE phage antibody library and the screening of the anti—trichosanthin IgE

    Institute of Scientific and Technical Information of China (English)



    Allergen specific IgE response is the major cause of immediate hypersensitivity.However the number of IgEproducing B cells and the amount of IgE,especially the specific IgE,are so low,it greatly impedes the study of the allergic-specifc antibody responses.Here we report the construction of a normal human IgE combinatorial library.The repertoire of IgE VH genes and of κ genes were separately amplified from normal human peripheral blood lymphocytes through RT-PCR,and were then constructed to form the phage surface display human Fab(IgEVH) library.A plant protein allergen,trichosanthin(TCS),was used to affinity-enrich and to screen the anti-TCS phage HuFab clones from the library.Human IgE(Fab) to TCS were detected.

  8. Chlamydia trachomatis Pgp3 Antibody Population Seroprevalence before and during an Era of Widespread Opportunistic Chlamydia Screening in England (1994-2012) (United States)

    Horner, Patrick J.; Craig, Rachel; Mindell, Jennifer S.; Murphy, Gary; McClure, Myra O.; Soldan, Kate; Nardone, Anthony; Johnson, Anne M.


    Background Opportunistic chlamydia screening of <25 year-olds was nationally-implemented in England in 2008 but its impact on chlamydia transmission is poorly understood. We undertook a population-based seroprevalence study to explore the impact of screening on cumulative incidence of chlamydia, as measured by C.trachomatis-specific antibody. Methods Anonymised sera from participants in the nationally-representative Health Surveys for England (HSE) were tested for C.trachomatis antibodies using two novel Pgp3 enzyme-linked immunosorbent assays (ELISAs) as a marker of past infection. Determinants of being seropositive were explored using logistic regression among 16–44 year-old women and men in 2010 and 2012 (years when sexual behaviour questions were included in the survey) (n = 1,402 women; 1,119 men). Seroprevalence trends among 16–24 year-old women (n = 3,361) were investigated over ten time points from 1994–2012. Results In HSE2010/2012, Pgp3 seroprevalence among 16–44 year-olds was 24.4% (95%CI 22.0–27.1) in women and 13.9% (11.8–16.2) in men. Seroprevalence increased with age (up to 33.5% [27.5–40.2] in 30–34 year-old women, 18.7% [13.4–25.6] in 35–39 year-old men); years since first sex; number of lifetime sexual partners; and younger age at first sex. 76.7% of seropositive 16–24 year-olds had never been diagnosed with chlamydia. Among 16–24 year-old women, a non-significant decline in seroprevalence was observed from 2008–2012 (prevalence ratio per year: 0.94 [0.84–1.05]). Conclusion Our application of Pgp3 ELISAs demonstrates a high lifetime risk of chlamydia infection among women and a large proportion of undiagnosed infections. A decrease in age-specific cumulative incidence following national implementation of opportunistic chlamydia screening has not yet been demonstrated. We propose these assays be used to assess impact of chlamydia control programmes. PMID:28129328

  9. Thyroid peroxidase antibodies in early pregnancy: utility for prediction of postpartum thyroid dysfunction and implications for screening. (United States)

    Premawardhana, L D K E; Parkes, A B; John, R; Harris, B; Lazarus, J H


    Thyroid peroxidase antibodies (TPOAb) in pregnancy are a marker for postpartum (PPTD) and long-term thyroid dysfunction, with variable sensitivity and specificity in PPTD prediction. To test its utility in prediction, we recruited 308 TPOAb-positive (147 developed PPTD (PPTD group) and 161 remained euthyroid [PPTE group]) and 102 TPOAb-negative women (none developed PPTD), in early pregnancy (median, 18; range, 9-19 weeks' gestation). TPOAb levels were higher in the PPTD group (median) (125.2 kIU/L; p < 0.001), and in its hypothyroid (162.4 kIU.; p < 0.0001), hyperthyroid (114.2 kIU/L; p < 0.007), and biphasic (105.1 kIU/L; p < 0.02) variants, compared to the PPTE group (66.7 kIU/L) The incidence of PPTD was significantly higher with TPOAb levels above 58.2 kIU/L (early pregnancy versus postpartum; relative risk, 1.37 [95% confidence interval [CI] 1.17-1.61] versus 0.78 [95% CI 0.5-1.2]) compared to levels below. The integrated postpartum TPOAb response was higher in the PPTD group (median) (159 kIU/L per week) and its variants (hypothyroid; 199 kIU/L per week; biphasic, 180 kIU/L per week; hyperthyroid, 120 kIU/L per week), compared to the PPTE group (86 kIU/L per week p < 0.004). Median early pregnancy TPOAb levels in the PPTD and PPTE groups correlated well with the postpartum antibody response (r = 0.58, p < 0.001). The sensitivity of TPOAb in PPTD prediction was 100% (early pregnancy and postpartum), specificity 62% (early pregnancy) versus 41% (postpartum) and positive predictive value 48% (early pregnancy and postpartum). The timing of TPOAb testing, the sensitive assay used and the absence of PPTD in TPOAb-negative subjects contributed to this high sensitivity. We recommend TPOAb in early pregnancy as a useful predictor of PPTD, particularly in populations where PPTD does not occur in TPOAb-negative women.

  10. Establishment of bacteria display technology for Fab antibody library screening%筛选Fab抗体库的细菌展示技术的建立

    Institute of Scientific and Technical Information of China (English)

    徐黎明; 尹成凯; 任桂萍; 田辉; 王雪苯; 丁良君; 李德山


    目的:利用NlpA蛋白的前六个氨基酸(CDQSSS)将抗体锚定在细菌内膜建立筛选Fabs抗体库的展示技术,为今后抗体的研发工作奠定基础.方法:从pNAD质粒中克隆出NlpA leader(含有CDQSSS序列)基因序列,利用相应的酶切位点将该序列插入pComb3表达载体中构建成用于展示Fab的重组质粒pBFD.将从pEAI质粒克隆得到的anti-human IL-1β抗体的重链Fab和全长轻链分别插入到NlpA leader和pelB leader( pComb3载体自带的果胶酶基因前导肽)的下游.将pBFD-Fab转入到E.coli DH5α中诱导表达,原生质球制备后,采用梯度浓度的抗原进行孵育,最后经流式细胞术(FCM)检测抗体展示情况并且分选阳性群体,利用质粒提取的方法来替代PCR方法拯救阳性基因,转化E.coli DH5α,利用FCM再次检测该群体展示的抗体与抗原结合情况.结果:所展示的anti-hlL-1β Fab抗体依次与抗原和FITC标记的抗原特异性抗体孵育后,用FCM实时检测,结果显示出很强的荧光信号并且表现出抗原浓度依赖性.拯救出的pBFD-Fab-原生质球的FCM检测结果与首次展示的FCM结果一致,该系统能够稳定的展示抗体.结论:经过该细菌展示系统展示的Fab抗体能够有效的折叠,与相应的抗原具有很好的特异性结合能力.成功改进该展示技术的基因拯救方法,避免了基因突变和链置换的发生.此外还证明了该展示技术具有很好的稳定性.本实验成功构建了筛选Fab抗体库的细菌展技术.%AIM:To establish bacterial display technology for the purpose of Fab antibody library screening, by u-sing six amino acids (CDQSSS) of the amino termimus of NlpA protein to anchore antibodies to the periplasmic side of the bacterial inner membrane. METHODS: The NlpA Leader sequences (encoding CDQSSS) was amplified from pNAD plasmid. The PCR product was subcloned into pComb3 expression vector to generate Fab display vector pBFD. The heavy chains of the Fab gene

  11. Presence of Autoimmune Antibody in Chikungunya Infection

    Directory of Open Access Journals (Sweden)

    Wirach Maek-a-nantawat


    Full Text Available Chikungunya infection has recently re-emerged as an important arthropod-borne disease in Thailand. Recently, Southern Thailand was identified as a potentially endemic area for the chikungunya virus. Here, we report a case of severe musculoskeletal complication, presenting with muscle weakness and swelling of the limbs. During the investigation to exclude autoimmune muscular inflammation, high titers of antinuclear antibody were detected. This is the report of autoimmunity detection associated with an arbovirus infection. The symptoms can mimic autoimmune polymyositis disease, and the condition requires close monitoring before deciding to embark upon prolonged specific treatment with immunomodulators.

  12. Analysis of HIV Antibody Screening in 9 844 STD Outpatients%性病门诊9844例患者HIV抗体筛查分析

    Institute of Scientific and Technical Information of China (English)

    陈龙庆; 杨欢; 张信江; 罗显华; 董泽令; 黄健


    Objective To understand the situation of the human immunodeficiency virus (HIV) infection in the sexually transmitted diseases (STD) outpatients.Methods HIV antibody was tested by the enzyme-linked immunosorbent assay (ELISA) in 9 844 cases of STD outpatients,positive serum was sent to the provincial CDC to confirm with Western blot test (Western-Blot,WB).Results In the 9 844 cases of STD outpatients,there were 71 cases with HIV positive antibody,the positive rate was 7.21‰ (71/9844).The age of major patien(t)s was 21 to 40 old years (55/71,77.46%).Although their occupational distribution involved civil servan,lawyers,students and high school students,businessmen,self-employed,hairdresser,car drivers,service personnel,sales personnel and the postman,etc.but no fixed occupational patients was more (42/71,59.15%).Conclusion The test of HIV antibody is an important way for finding HIV/AIDS patients.The HIV/AIDS patients may be detect with HIV antibody screening in general hospital.It have a very important significance to cut transmission and avoid iatrogenic transmission of HIV/AIDS.%目的 了解性病(STD)门诊患者中人类免疫缺陷病毒(HIV)感染情况.方法 采用酶联免疫吸附试验(ELISA法)对9 844例STD门诊患者进行HIV抗体检测,阳性者送省CDC做蛋白印迹试验(Western-Blot,WB)确诊.结果 9 844例STD门诊患者中,HIV抗体阳性者71例,阳性检出率7.21‰(71/9 844).年龄分布以中青年为主,21 ~40岁年龄组占77.46% (55/71);职业分布虽涉及公务员、律师、学生、商人、个体户、美发师、汽车司机、服务人员、销售人员与邮递员等,但以无固定职业者为多(42/71例,59.15%).结论 对综合医院STD门诊患者进行HIV抗体筛查是发现HIV/AIDS患者的重要途径,应加强对性病患者的HIV抗体筛查,以便及时发现HIV/AIDS患者,这对切断HIV/AIDS患者的传染途径与避免医源性传播都有十分重要的意义.

  13. Anti-DNA antibodies--quintessential biomarkers of SLE. (United States)

    Pisetsky, David S


    Antibodies that recognize and bind to DNA (anti-DNA antibodies) are serological hallmarks of systemic lupus erythematosus (SLE) and key markers for diagnosis and disease activity. In addition to common use in the clinic, anti-DNA antibody testing now also determines eligibility for clinical trials, raising important questions about the nature of the antibody-antigen interaction. At present, no 'gold standard' for serological assessment exists, and anti-DNA antibody binding can be measured with a variety of assay formats, which differ in the nature of the DNA substrates and in the conditions for binding and detection of antibodies. A mechanism called monogamous bivalency--in which high avidity results from simultaneous interaction of IgG Fab sites with a single polynucleotide chain--determines anti-DNA antibody binding; this mechanism might affect antibody detection in different assay formats. Although anti-DNA antibodies can promote pathogenesis by depositing in the kidney or driving cytokine production, they are not all alike, pathologically, and anti-DNA antibody expression does not necessarily correlate with active disease. Levels of anti-DNA antibodies in patients with SLE can vary over time, distinguishing anti-DNA antibodies from other pathogenic antinuclear antibodies. Elucidation of the binding specificities and the pathogenic roles of anti-DNA antibodies in SLE should enable improvements in the design of informative assays for both clinical and research purposes.

  14. The significance of irregular antibody screening among healthy blood donors for clinical blood transfusion%健康献血者不规则抗体筛查在临床输血中的意义

    Institute of Scientific and Technical Information of China (English)

    吕学琴; 尹向丽


      目的探讨健康献血者不规则抗体筛查在临床输血中的意义。方法收集2011年8月至2012年10月3029名健康献血者的血液标本,采用盐水介质法和抗人球蛋白法进行不规则抗体筛查,抗体筛查阳性者做进一步鉴定,分析抗体特异性。结果对新疆维吾尔族自治区农三师图木舒克市中心血站3029例健康献血者进行不规则抗体筛查阳性17例(0.56%),其中抗-D阳性9例,抗-E阳性4例,抗-e阳性1例,抗-M阳性2例,抗A1阳性1例。男性不规则抗体阳性率为0.43%(6/1397),女性不规则抗体阳性率为0.67%(11/1632),差异无统计学意义(P>0.05)。有输血史或妊娠史的健康献血者不规则抗体阳性率显著高于无输血史或妊娠史者(1.48% vs 0.10%,P<0.05)。结论常规对健康献血者血液标本进行不规则抗体筛查和鉴定,对保障临床用血安全、预防溶血性输血反应具有极其重要的意义。%Objective To investigate the significance of screening irregular antibodies among healthy blood do-nors for clinical blood transfusion .Methods 3 029 blood samples of healthy donors from Augst 2011 to October 2012 were collected .Saline medium method and Coombs test were used for irregular antibodies screening .The positive samples of the screening tests were further evaluated to analyze their antibody specificity .Results Among the 3 029 healthy donors ,17 cases (0 .56% ) were positive for irregular antibodies screening ,including 9 cases for anti-D anti-body ,4 cases for anti-E antibody ,1 case for anti-e antibody ,2 cases for anti-M antibody and 1 case for anti-A1 anti-body respectively .The positive rates of male and female were 0 .43% (6/1 397) and 0 .67% (11/1 632) .The differ-ence had no statistical significance (P>0 .05) .The positive rate of irregular antibody in healthy donors who had his-tory of blood transfusion or pregnancy was significantly higher

  15. A major Sm epitope anchored to sequential oligopeptide carriers is a suitable antigenic substrate to detect anti-Sm antibodies. (United States)

    Petrovas, C J; Vlachoyiannopoulos, P G; Tzioufas, A G; Alexopoulos, C; Tsikaris, V; Sakarellos-Daitsiotis, M; Sakarellos, C; Moutsopoulos, H M


    A sensitive, highly reproducible, solid-phase enzyme immunoassay (ELISA), was developed in order to investigate whether the synthetic heptapeptide PPGMRPP-a major epitope of the Sm autoantigen-anchored in five copies to a sequential oligopeptide carrier (SOC), [(PPGMRPP)5-SOC5] is a suitable antigenic substrate to identify anti-Sm/antibodies. Sera with different autoantibody specificities [45 anti-Sm, 40 anti-U1RNP, 40 anti-Ro (SSA)/La(SSB) positive, 21 Antinuclear antibody positive, but negative for antibodies to extractable nuclear antigens (ANA + /ENA - ) and 75 normal human sera, ANA negative] and 75 sera from patients with rheumatoid arthritis (RA) were tested for anti-(PPGMRPP)5-(SOC)5 reactivity in order to evaluate the specificity and sensitivity of the method to detect anti-Sm antibodies. RNA immunoprecipitation assays for the detection of anti-Sm and anti-U1RNP antibodies and counter immunoelectrophoresis (CIE) for the detection of anti-Ro(SSA) and anti-La(SSB) antibodies were used as reference techniques. The sensitivity of the method was 98% and the specificity was 68% for the determination of anti-Sm antibodies, while for the determination of anti-Sm and/or anti-U1RNP reactivity (antibodies to snRNPs) the corresponding values were 82% and 86%, respectively. In a comparison of the above assay with an ELISA, using Sm/U1RNP purified complex as immobilized antigen it was shown that the sensitivity of the anti-Sm/U1RNP ELISA in detecting anti-snRNPs was 74%; in addition sera with anti-Sm antibodies gave higher binding in the anti-(PPGMRPP)5-(SOC)5 ELISA compared with anti-Sm/U1RNP ELISA. Intra- and inter-assay precision was measured on four sera with reactivities extending into a wide range of absorbance values showed that the intra-assay coefficient of variation (CV%) ranged from 2.7 to 6 and the inter-assay CV% ranged from 9 to 14.5. These results indicate that the PPGMRPP peptide anchored to a pentameric SOC as a carrier is a suitable antigen for

  16. Screening and immunological identification of the human ScFv antibody against PSMA%全人源抗PSMA单链抗体的筛选及免疫活性鉴定

    Institute of Scientific and Technical Information of China (English)

    张才田; 刘金霞


    Objective To screen and identify a human single-chain variable fragment(scFv) antibody against prostate specific membrane antigen(PSMA) from a human scFv antibody library.Methods Using a synthetic PSMA peptide as the coating antigen, the antibody library was screened by five rounds of combining-eluting-amplification.The phage antibody against PSMA with high specificity was screened out from the human scFv antibody library and its binding ability to the antigen was tested by ELISA.The soluble antibody was produced by plasmids extracted from highly specific clones, whose binding ability to PSMA was further identified by Western blot and immunohistochemistry.The affinity constant of the soluble antibody was measured by non-competitive ELISA.Results The screened phage antibody was specific for PSMA by ELISA.The soluble antibody was also specific for PSMA, its molecular weight was about 30 kD by SDS-PAGE and its affinity constant was about 5.077 × 106 L/mol.Conclusions The screened scFv antibody is specific and has low immunogenicity.It can be further used in the target treatment of malignant tumors.%目的 从全人源单链噬菌体抗体库中筛选出抗前列腺特异性膜抗原(PSMA)特异性单链抗体并进行免疫活性鉴定.方法 以合成的PSMA多肽为抗原,经过五轮吸附-洗脱-扩增,从单链噬菌体抗体库中筛选出特异性抗PSMA噬菌体抗体,ELISA检测其抗原结合能力,并对特异性较强的克隆提取质粒,表达可溶性抗体.Western Blotting和免疫组织化学检测其抗原结合性,非竞争ELSIA法检测其亲和常数.结果 从单链噬菌体抗体库中筛选出的噬菌体抗体,经ELISA鉴定为抗PSMA的特异性噬菌体抗体.抗PSMA可溶性抗体相对分子质量约为3.0×104,与PSMA特异性结合,其亲和常数约为5.077×106L/mol.结论 所得全人源抗PSMA单链抗体保留完整抗体分子结合抗原的特异性,免疫原性弱,是肿瘤导向治疗的理想栽体.

  17. A simplified method to attach antibodies on liposomes by biotin-streptavidin affinity for rapid and economical screening of targeted liposomes. (United States)

    Papadia, Konstantina; Markoutsa, Eleni; Antimisiaris, Sophia G


    The biotin-Streptavidin (STREP) technique for attachment of monoclonal antibodies (mAbs) (or other ligand types) on liposome surface offers high attachment yield, however it is time consuming and expensive due to the number of steps used and the consumption of large quantities of STREP. Herein, a simplified, fast and economic technique, by incubating pre-mixed biotin-mAb/STREP with biotin-liposomes, at a 3:1:1 biotin-mAb/STREP/biotin-LIP ratio (mol/mol/mol) was evaluated. The physichochemical properties, final mAb attachment yield and targeting potential of liposomes decorated with an anti-transferrin receptor mAb (TfR-mAb), prepared by the simple method (SM) and the conventional method (CM), were compared. The vesicle uptake by hCMEC/D3 cells (known to overexpress TfR) were considered as a measure of liposome targeting capability. Results show that both targeted liposome types (SM and CM) have small size (mean diameters around 150 nm), low poly-dispersity (approx. 0.20) and similar mAb attachment yield (between 64-88%). However, the uptake of the SM-liposomes is slightly lower compared to CM-LIP (24-30% decrease), suggesting that the modulated conformation of mAbs on the liposome surface (triplets attached to one single STREP molecule) results in decreased targeting capability. Nevertheless, the simpler and faster one-step preparation procedure which has very high lipid recovery (> 95%) compared to the CM (50-60%) and 15-30 times lower consumption of STREP, may be a good alternative for initial screening of various mAbs as ligands for targeted liposomal or other nanotechnologies, during pre-clinical development.

  18. Detection of filaria-specific IgG4 antibodies and filarial DNA, for the screening of blood spots for Brugia timori. (United States)

    Fischer, P; Bonow, I; Supali, T; Rückert, P; Rahmah, N


    The establishment of simple, sensitive and specific tools for the diagnosis of brugian lymphatic filariasis is a prerequisite for a successful intervention to control the disease. In the simple and rapid Brugia Rapid (BR) test, an immunochromatographic dipstick is used to detect IgG(4) antibodies that are reactive with a recombinant Brugia malayi antigen. When sera from 109 individuals with Brugia microfilaraemias (12 with B. malayi and 97 with B. timori) were investigated using the BR test, all were found positive. In contrast, all of the 150 sera from individuals with Onchocerca volvulus or Mansonella infections investigated were found negative in BR tests. Some unwelcome cross-reactions were observed, however, with sera from individuals infected with Wuchereria bancrofti (three of 12 test-positive) and Dirofilaria (one of nine test-positive). In an attempt to facilitate sample collection and detect any cross-reactions, the BR dipstick was used to screen blood spots, that had been allowed to dry on filter paper, for B. timori microfilariae, before the dipstick-positive samples were tested with a PCR-based assay. Of the 66 individuals so tested, 37 (56%) were found positive by the BR test used on dry blood spots and eight (22%) by the filtration of fresh blood samples. Only nine of the 37 dipstick-positive samples were found PCR-positive. The combined use of BR tests and PCR-based assays, for testing blood spots in areas where brugian filariasis is endemic, appears to be a promising method not only for post-treatment monitoring but also for the certification activities planned within the framework of the Global Programme to Eliminate Lymphatic Filariasis.

  19. Computational Screening of the Human TF-Glycome Provides a Structural Definition for the Specificity of Anti-Tumor Antibody JAA-F11


    Tessier, Matthew B.; Oliver C Grant; Jamie Heimburg-Molinaro; David Smith; Snehal Jadey; Gulick, Andrew M.; John Glushka; Deutscher, Susan L.; Kate Rittenhouse-Olson; Woods, Robert J


    Recombinant antibodies are of profound clinical significance; yet, anti-carbohydrate antibodies are prone to undesirable cross-reactivity with structurally related-glycans. Here we introduce a new technology called Computational Carbohydrate Grafting (CCG), which enables a virtual library of glycans to be assessed for protein binding specificity, and employ it to define the scope and structural origin of the binding specificity of antibody JAA-F11 for glycans containing the Thomsen-Friedenrei...

  20. Screening for epitope specificity directly on culture supernatants in the early phase of monoclonal antibody production by an ELISA with biotin-labeled antigen

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Jensen, Charlotte H; Gregersen, Annemette;


    This report describes an assay for comparison of epitope specificity in groups of monoclonal antibodies against a given antigen. The only prerequisite is the biotin-labeled antigen. One of the monoclonal antibodies is captured onto a plastic surface via a rabbit anti-mouse Ig, and the other...... preincubated with biotinylated antigen. When the two antibodies react with the same epitope subsequent binding of the biotin-labeled antigen is abolished (inhibition). In the cases where no inhibition was observed, the two antibodies were considered to react with distinct, independent epitopes. The obvious...

  1. 胶体金法快速筛查HIV抗体的应用研究%The Study of Practical HIV-Antibody for Rapid Screening with Colloidal Gold Method

    Institute of Scientific and Technical Information of China (English)

    李雪; 姚勇; 漆兆李; 蒋玲


    Objectives This article is aimed to study the adaptability and feasibility of HIV-antibody for rapid screening with colloidal gold method. Methods Compared with the method of ELISA to the blood sample for HIV-antibody, we collected the sample from the exudation of the oral mucosa with colloidal gold method for rapid screening to the HIV-antibody. Results All cases of 150, we had analyzed with the method of ELISA and colloidal gold for the HIV-antibody, which were negative in 127 cases (84.7%) and positive in 23 cases (15.3%) in rapid screening with colloidal gold method for the exudation of oral mucosa while were negative in 129 cases (86.0%) and positive in 21 cases (14.0%) in the method of ELISA for the blood samples, found there is no obvious difference (P>0.05). Conclusion The method of rapid screening HIV-antibody with colloidal gold for the exudation of oral mucosa is worth to popularize and is feasibility in the basic medical unit.%  目的探讨胶体金法快速筛查 HIV 抗体的可行性和适用性。方法用胶体金法快速筛查测试对象口腔黏膜渗出液中 HIV 抗体,与ELISA 法检测相同测试对象血样 HIV 抗体作对照。结果本组研究对象150例,用胶体金法快速筛查其口腔黏膜渗出液样本的 HIV 抗体,阴性127例(阴性率84.7%),阳性23例(阳性率15.3%);再用 ELISA 法检测研究对象的血浆标本的 HIV 抗体,阴性129例(阴性率86.0%),阳性21例(阳性率14.0%),二者结果无显著差异(P >0.05)。结论用胶体金法检测人类口腔黏膜渗出液样本作为快速筛查HIV 抗体是可行的,值得基层医疗机构推广应用。

  2. Analysis of 88 Cases of Elderly Patients with Treponema Pallidum Antibody Positive Screening Results%88例老年患者梅毒螺旋体抗体筛查阳性结果的分析

    Institute of Scientific and Technical Information of China (English)

    杨清梅; 贾伟建


    Objective To investigate the value of Treponema pallidum antibody screening positive for elderly patients. Method The detection of Treponema pallidum antibody in the serum of 3520 elderly patients by chemiluminescence method, and then the positive results of Treponema pallidum gelatin agglutination test (TPPA) and toluidine red unheated serum test (TRUST) for a test, antibody indeterminate samples by immunoblotting of Treponema pallidum (Western blot, WB) method for the final confirmation. Results The chemiluminescence method screening of 88 cases of syphilis antibody positive, confirmed by TPPA positive 63 cases, TRUST was positive in 17 cases, which were positive for TPPA validation. 25 cases were identified by uncertainty, WB was positive in 1 cases, 7 cases of suspected, negative in 17 cases. Conclusion Elderly patients with Treponema pallidum antibody screening will have certain false positive, final confirmation of a single screening positive samples by Western blotting.%目的:探讨梅毒螺旋体抗体筛查阳性对老年患者的价值。方法采用化学发光法检测3520例老年患者血清中梅毒螺旋体抗体,阳性结果再用梅毒螺旋体明胶凝集试验(TPPA)和甲苯胺红不加热血清试验( TRUST)进行检测,对不确定标本再用免疫印迹(Western blot, WB)法进行最后确认。结果化学发光法共筛查出88例梅毒螺旋体抗体阳性,经TPPA确证阳性63例,TRUST阳性17例,当中TPPA确证均阳性。25例不确定标本,WB法确认1例阳性,7例可疑,17例阴性。结论老年患者梅毒螺旋体抗体筛查会有一定的假阳性,对单筛查阳性的标本应用免疫印迹法作最后确认。

  3. Rapid Screening for Potential Epitopes Reactive with a Polycolonal Antibody by Solution-Phase H/D Exchange Monitored by FT-ICR Mass Spectrometry (United States)

    Zhang, Qian; Noble, Kyle A.; Mao, Yuan; Young, Nicolas L.; Sathe, Shridhar K.; Roux, Kenneth H.; Marshall, Alan G.


    The potential epitopes of a recombinant food allergen protein, cashew Ana o 2, reactive to polyclonal antibodies, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Ana o 2 polyclonal antibodies were purified in the serum from a goat immunized with cashew nut extract. Antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:polyclonal antibody (Ag:pAb) complexes. Complexed and uncomplexed (free) rAna o 2 were then subjected to HDX-MS analysis. Four regions protected from H/D exchange upon pAb binding are identified as potential epitopes and mapped onto a homologous model.

  4. 211例血清抗核抗体及抗核抗体谱检测结果联合应用的分析%Joint detection of serum antinuclear antibodies and antinuclear antibody spectrum in 211 cases

    Institute of Scientific and Technical Information of China (English)

    李彩虹; 张振国; 乐宏元


    目的 探讨抗核抗体(ANA)与特异性抗核抗体谱(ANAs)检测结果联合应用的临床意义.方法 采用间接免疫荧光法(IIF)筛查211份临床送检血清标本的ANA,采用条带酶免分析法(LIA)检测ANAs,分析检测结果联合应用的临床意义.结果 211例标本血清IIF-ANA +/HA-ANAs+患者有65例,IIF-ANA+/HA-ANAs-患者有9例,IIF-ANA-/HA-ANAs+患者有5例,IIF-ANA-/LIA-ANAs-患者有132例,总符合率为93.37%.结论 IIF-ANA筛查和LIA-ANAs检测不能相互代替,两者同时检测可提高检出率.

  5. Comparative analysis of antinuclear antibody and antinuclear antibody spectrum for 2 513 samples%2513例抗核抗体与抗核抗体谱检测结果对比分析

    Institute of Scientific and Technical Information of China (English)

    蒲泽晏; 胥国强; 李祥坤; 李凤; 黄雪梅


    目的 分析本实验室抗核抗体(ANA)与抗核抗体谱(ANAs) 15项联合检测的结果,并探讨两者之间的关系.方法 分别用间接免疫荧光法(IIF)和免疫印迹法(IBT)检测患者ANA和ANAs,判断其ANA滴度、荧光模型和ANAs结果,并对ANA和ANAs检测结果进行对比分析.结果 471例ANA阳性患者标本中,颗粒型161例,均质型110例,胞质型96例、核仁型37例、着丝点型28例,其他型合计39例;滴度为1∶100者196例,占41.61%(196/471),其中ANAs阳性53例,占27.04%(53/196);滴度为1∶320者123例,占26.11%(123/471),其中ANAs阳性67例,占54.47%(67/123);滴度为1∶1 000者152例,占32.27%(152/471),其中ANAs阳性132例,占86.84%(132/152);2 042例ANA阴性标本中45例ANAs检测阳性,阳性率为2.2%(45/2042);高滴度ANA患者血清中ANAs的阳性率高于低滴度(x2=123.132,P<0.05).结论 不同滴度的ANA与ANAs之间有一定的关系,如果以ANA进行过筛,阳性再做ANAs会漏检部分患者,建议IIF和IBT联合应用,防止风湿病的漏诊.

  6. Clinical Evaluation of BioPlex 2200 HIV Ag-Ab, an Automated Screening Method Providing Discrete Detection of HIV-1 p24 Antigen, HIV-1 Antibody, and HIV-2 Antibody


    Salmona, Maud; Delarue, Severine; Delaugerre, Constance; Simon, François; Maylin, Sarah


    Early and accurate diagnosis is essential for optimal therapeutic outcomes in patients infected with HIV. Currently, none of the commercially available fourth-generation assays differentiate HIV-1 and HIV-2 antibodies (Ab) or the HIV-1 p24 antigen (Ag). The aim of this study was to evaluate the performance of a novel assay, the BioPlex 2200 HIV Ag-Ab. This assay uses a multiplex flow immunoassay design allowing the simultaneous detection and identification of antibodies to HIV-1 (groups M and...

  7. 抗体筛查细胞抗原涵盖范围与电子配血技术的安全应用%Application of antibody screening cell antigen coverage and electronic matching of blood

    Institute of Scientific and Technical Information of China (English)

    刘家瑞; 王远杰; 冯宁; 李红梅; 冯俊程


    Objective To study the significance of antibody screening cell antigen range in the electronic matching technology security application.Methods ABO/RhD of blood donor and blood recipient were detect with blood type and antibody.The blood matching were used by electronic matching technology and polybrene crossmatch.Results Of 22,790 blood samples of patients,there were 164 cases with positive antibody screening (0.72%).Of 15740 blood donor blood samples,there were 11 cases with positive antibody screening (0.07 %).Of 22,540 blood samples in line with the rules of the electronic distribution of blood,the implementation of electronic matching of blood ABO / RhD blood group incompatibility was not found by the computer.Of 22534 blood samples with blood compatible,6 blood samples polybrene crossmatch was incompatible with the main side and found no secondary side incompatible.6 blood samples spectrum cell antibody specific identification of all the MNS system anti-Mur,missed rate of 0.027%.Conclusion Clinical antibody screening cells only serological crossmatch,whose antibody missing rate is relatively high,which cannot fully protect the security of the technology of the electronic matching of blood.%目的 探讨抗体筛查细胞抗原涵盖范围在保障电子配血技术安全应用中的意义.方法 分别对供受血者进行ABO/RhD血型检测以及抗体筛查.患者二次血型鉴定结果一致,抗体筛查结果阴性,按电子配血规则进行电子配血,同时采用凝聚胺法对供受血者进行传统血清学交叉配血平行试验,分析抗体筛查细胞抗原涵盖范围与不规则抗体漏检情况以及对电子配血技术安全应用的影响.结果 22790份患者血样,抗体筛查阳性164例,阳性率0.72%.15740份供血者血样,抗体筛查阳性11例,占0.07%.22540份血样均符合电子配血规则,由计算机实施电子配血未发现ABO/RhD血型不相容.用凝聚胺法进行传统血清学

  8. 清远市273份HIV抗体初筛阳性标本与确证结果分析%Analysis on preliminary screening and confirmed results in 273 HIV antibody positive samples in Qingyuan City

    Institute of Scientific and Technical Information of China (English)

    邓远玲; 詹巧莉; 罗文玲; 朱劲涛


    目的 了解清远市人类免疫缺陷病毒(HIV)抗体初筛实验与确证实验结果符合率,为提高实验室HIV抗体检测技术提供依据.方法 对清远市2010-2011年经酶联免疫吸附试验(ELISA)初筛的273份标本HIV抗体阳性及可疑阳性的结果与确证实验的结果进行比较分析.结果 273份标本,经蛋白印迹(WB)试验确证HIV-1阳性257例,占94.14%;不确定9例,占3.3%;阴性7例,占2.56%.筛查阳性与确证阳性总体符合率为94.14%,确证条带gp160、gp120、p24出现率均有95%以上;出现8条以上条带的有166份,出现率为64.59%.结论 初筛试剂检测HIV抗体与WB确证试验的检测结果符合率较高,但判断HIV抗体阳性或艾滋病必须经WB试验确证,以排除假阳性.%[ Objective]To understand the coincidence rate of HIV antibody screening and confirmed test in Qingyuan City, provide a basis for improving the HIV antibody examination technology. [ Methods ] The ELISA results of preliminary screening (273 HIV antibody positive samples and probable positive samples) in Qingyuan City from 2010-2011 were compared with the results of confirmed test. [Results]In 273 HIV antibody positive samples of screening, 257 cases were confirmed by Western blot (WB) test, which accounted for 94.14% , while 9 (3. 3% ) cases were uncertain and 7 (2. 56% ) cases were negative. The total coincidence rate of screening and confirmed test for positive cases was 94. 14%. The occurrence rate of gpl60, gpl20 and p24 reactive bands was over 95%. 166 cases had more than 8 bands in WB test (64. 59% ). [ Conclusion] The coincidence rate of HIV antibody screening and confirmed WB test is high. However, HIV antibody positive cases and AIDS cases must be confirmed by WB test to exclude false positive reaction.

  9. Clinical Significance of Screening of Rh-negative Blood Group and Detection of Irregular Antibody%Rh阴性血型筛查及不规则抗体检测的临床意义

    Institute of Scientific and Technical Information of China (English)

    李岚; 伍昌林; 党鑫堂; 董洪强; 朱奕


    目的 通过Rh阴性血型筛查及不规则抗体的检测,对抗体阳性的Rh阴性患者选择相合的血液输注,并分析其临床意义.方法 采用微柱凝集法对363例RhD阴性输血患者进行抗体筛查及Rh血型鉴定,选择Rh因子相合的血液输注,对有抗-c和抗-e抗体Rh阴性患者在血源紧缺的情况下,选择O型ccdEE和CCdee表型红细胞输注,观察临床输血效果.结果 在363例RhD阴性患者中,不规则抗体阳性21例,其中抗-D抗体5例,抗-E抗体8例,抗-c抗体3例,抗-c、E抗体2例,抗-C抗体2例,抗-e抗体1例,对存在不规则抗体的患者选择Rh因子相合的血液输注,临床效果良好.结论 根据抗体筛查及Rh血型鉴定结果,选择Ph因子相合的血液输注,可避免不规则抗体的产生,提高临床输血的有效性,减少输血不良反应的发生.%Objective To choose the matching blood to Rh-negative patients who were positive for irregular antibody by the screening of Rh-negative blood group and the detection of irregular antibody, and analyze its clinical significance.Methods A total of 363 RhD-negative patients were subjected to antibody screening and Rh blood grouping by micro-column agglutination method,based on which the Rh factor-matched blood was selected for transfusion.For lack of matched blood source, the RhD-negative patients with anti-c and anti-e antibodies were transfused with erythrocytes group O of ccdEE and CCdee phenotypes.Results Of the 363 RhD-negative patients, 21 were positive for irregular antibody, including 5 for anti-D, 8 for anti-E, 3 for anti-c, 2 for anti-c and E,2 for anti-C and 1 for anti-e.The patients with irregular antibodies were transfused with selected Rh factor-matched blood, and sarisfactory clinical efficacy was observed.Conclusion By transfusion with Rh factor-matched blood selected based on antibody screening and Rh blood grouping, the production of irregular antibodies was avoided, the efficacy of clinical blood transfusion was

  10. The role of HLA antibodies in allogeneic SCT: is the 'type-and-screen' strategy necessary not only for blood type but also for HLA? (United States)

    Yoshihara, S; Taniguchi, K; Ogawa, H; Saji, H


    The role of HLA antibodies in SCT has drawn increasing attention because of the significantly increased number of patients who receive HLA-mismatched SCT, including cord blood transplantation, haploidentical SCT and unrelated SCT. Technical advancements in the methods of HLA Ab testing have realized rapid, accurate and objective identification, as well as quantification of specific HLA antibodies. Recent clinical studies have suggested that the presence of donor-specific HLA antibodies (DSA) in patients is associated with graft failure in HLA-mismatched SCT when the above-listed stem cell sources are used and results in different impacts. Of note, most of the 'HLA-matched' unrelated SCT actually involve HLA mismatches in HLA-DP and the presence of antibodies against this locus has been reported to be associated with graft failure. Thus, HLA Ab should be examined as a work-up for all patients who undergo SCT from 'alternative donors.' The simplest route for preventing HLA Ab-mediated graft failure in Ab-positive patients is to avoid donors who possess the target Ag of HLA antibodies. If SCT from such donors must be performed, treatment for DSA before SCT may improve the chances of successful donor engraftment.

  11. Influence of clinical features, serum antinuclear antibodies, and lung function on survival of patients with systemic sclerosis

    DEFF Research Database (Denmark)

    Jacobsen, Søren; Ullman, S; Shen, G Q;


    of all new patients with SSc. Disease onset was defined as the time of onset of cutaneous sclerosis. Vital status and causes of death were determined at the end of the observation period. Data on clinical status and pulmonary function were obtained. Antitopoisomerase I (anti-topo I), anticentromere, anti...... of 13.3 yrs; 16 died of an SSc related condition and 50 of other causes. Pulmonary fibrosis, DLCO reduction

  12. Antimitochondrial antibody (United States)

    ... page: // Antimitochondrial antibody To use the sharing features on this page, please enable JavaScript. Antimitochondrial antibodies (AMA) are substances ( antibodies ) that form against mitochondria. ...

  13. Analysising the result between HIV antibody screening laboratories in Liuzhou%2012年柳州市HIV抗体筛查实验室比对结果分析

    Institute of Scientific and Technical Information of China (English)

    陈威; 潘洁; 陈柳军; 杨振兴; 刘鑫


    目的 通过组织柳州市县HIV抗体筛查实验室进行一次室间比对,提高各个HIV抗体筛查实验室检测能力,减少其在日常检测中的差错.方法 通过下发盲样的方式,共36家HIV抗体筛查实验室,均采用ELISA的方法对考核标本(QC001、QC002、QC003、QC004、QC005)进行单独检测,并及时反馈结果.结果 36家实验室对5个盲样的检测结果显示,除了QC005的正确率为97.2%外,其他几个盲样的考评正确率均达100.0%,分析定量检测结果显示CV值波动较大,只有个别盲样CV值能控制在25%内.结论 全市HIV抗体筛查实验室建设已经初具规模并且有良好的检测能力,但质量控制仍需加强.%Objective To evaluate the level of test and standardize management in HIV antibody screening laboratories in Liuzhou city in order to promote them to improve their whole test ability and reduce the error in the routine detection.Methods Having 36 HIV antibody screening laboratories joined the examination.Them using the method of ELISA to detect these blind samples(QC001,QC002,QC003,QC004,QC005),then returned the result to us.Result The results of blind samples showed that the coincidence rates all were 100% but the QC005 was 97.2%.Analysising the quantitative testing results showed that CV value was fluctuant greatly,shows that the HIV antibody screening laboratory quality control should be improved.Conclusion The overall quality of the HIV screening laboratories in Liuzhou city is satifactory but need to enhance in the quality control.

  14. Comprehensive evaluation of two HLA-B17 monoclonal antibodies for flow cytometry-based HLA-B57/B58 screening prior to abacavir prescription. (United States)

    Stevens, R; Coates, E; Street, J; Cook, E; Darke, C


    Hypersensitivity reactions to the drug abacavir, used to treat HIV/AIDS patients, is associated with possession of HLA-B*57:01. We have carefully assessed two commercially available HLA-B57/B58 murine monoclonal antibodies [0196HA and BIH0243 (One Lambda Inc.)] in a simple flow cytometry-based assay. The evaluation involved tests on 228 reference and random samples covering 91% of all WHO recognized HLA-A, B and C specificities. These involved donors with six different HLA-B*57 alleles and included 19 examples of B*57:01. Both antibodies unambiguously detected B57, but there were small difference in their reactivity against B57-positive non-B*57:01 samples. Importantly, there was no reactivity against B57/B58-negative samples. The possible amino acid motifs involved in the reactivity of these antibodies with B57/B58 were delineated. Thus, HLA-B57/B58, normally present in <10% of patients, can be easily recognized using these two antibodies and further tested by a DNA-based typing method to identify B*57:01.

  15. The high incidence of anti-Ro/SSA and anti-p200 antibodies in female patients with connective tissue diseases confirms the importance of screening for congenital heart block-associated autoantibodies during pregnancy. (United States)

    Cozzani, E; Agnoletti, Arianna Fay; Pappalardo, F; Schiavetti, I; Torino, A; Parodi, A


    It is known that anti-Ro/SSA positivity leads to higher risk of miscarriage and fetal cardiac malformations. Particularly, anti-p200 antibodies against a finer specificity of the Ro/SSA antigen, have been associated with congenital heart block. The aim of the study was to assess the frequency of anti-p200 among female patients with different connective tissue diseases and, among these, the relevance of anti-p200 values in patients with cutaneous diseases compared to systemic diseases. Anti-p200 were investigated in 110 anti-Ro/SSA positive female sera, sent to our laboratory between 2008 and 2014 with suspect of connective disease, by using ELISA testing. Positivity was found in 40.9 % samples, 34 of them showed a strong positivity (values ≥ 1.0, cut off = 0.7). Patients with systemic diseases were anti-p200 positive in the 45.9 % of cases while patients with cutaneous diseases were positive in the 24.0 % of cases. Positivity for anti-p200 antibodies was revealed in 24.0 % of patients with discoid lupus erythematosus; 100 % of patients with dermatomyositis; 40.0 % of patients with mixed connective tissue disease; 25.0 % of patients with rheumatoid arthritis; 100 % of patients with Sjögren's syndrome; 33.3 % of patients with subacute cutaneous lupus erythematosus; 42.9 % of patients with systemic lupus erythematosus; 80.0 % of patients with systemic sclerosis. No significant difference in anti-p200 prevalence was found between systemic and cutaneous involvement, nevertheless, considering only positive sera, the antibody titer was higher in systemic diseases rather than in cutaneous diseases (2.6 ± 1.7 and 1.7 ± 1.9; p = 0.041). The authors think screenings for anti-Ro/SSA and anti-p200 antibodies should be included in the laboratory checklist for pregnancy.

  16. Rescreening and confirmation of anti-HIV antibody positive serum in primary screening in Qingdao%青岛市HIV抗体初筛阳性标本的复检及确证结果分析

    Institute of Scientific and Technical Information of China (English)

    史晓燕; 张洪花; 赵国有; 弋英; 汪照国


    Objective To rescreen and confirm anti-HIV antibody positive serum in primary screening in Qingdao from 2009 to 2011 and to analyze the related data,so as to provide basis for prevention and control of AIDS in Qingdao.Methods Samples positive for anti-HIV antibody in primary screening test were re-screened with both Electroselenium method and ELISA.Then the positive samples in either re-screening test were further confirmed with Western Blot (WB).Results In 1348 positive samples from primary screening test,positive were found in 745 cases in re-screening test.By WB confirmation,563 cases were positive,95 cases were suspected and 87 cases were negative.In contrast to WB,positive coincidence rates of ELISA and Electroselenium method were 81.5% and 84.3% respectively.The sensitivity and specificity of ELISA were 100% and 93.6 % respectively,and those of Electroselenium method were 100% and 94.5% respectively.Flouting population accounted for 58.4% and MSM accounted for 40.1% of total HIV infection.Conclusion Anti-HIV antibody rescreening tests could exclude most false positive samples in primary screening test.However,there is still some false positive rate in either rescreening test.Flouting population and MSM are the high risk population of HIV infection in Qingdao city and intervention should be strengthened among those people.%目的 复检和确认HIV抗体初筛阳性血清并分析相关资料,为青岛市艾滋病防治工作提供依据.方法 对初筛阳性血清,用胶体硒和ELISA试验复检,任一复检试验阳性的血清再用WB试验确证并对确证阳性病例进行流行病学分析.结果 1348份初筛阳性血清经复检745份阳性,WB确证563份HIV-l抗体阳性,不确定95份,阴性87份.ELISA、胶体硒法与WB的阳性符合率分别为81.5%和84.3%,二者的敏感度均为100%,特异度分别为93.6%和94.5%,确证阳性人群中,外来人员占58.4%,男男性行为人群占40.1%.结论 ELISA和胶体硒

  17. Nuclear energy in postwar Japan and anti-nuclear movements in the 1950s. (United States)

    Yamazaki, Masakatsu


    The atomic bombings of Hiroshima and Nagasaki in August 1945 revealed the most destructive power to-date of man-made weapons. Their impact was so great that Japanese scientists thought that a bigger disaster could be prevented only if war was abolished. Thus they welcomed the international control of atomic energy. It was, however, only after the occupation that the Japanese general public began to learn about the horror of these atomic disasters due to the censorship imposed by the occupational forces. The hydrogen bomb test by the US in the Bikini atoll on March 1, 1954 renewed fears of nuclear weapons. The crew of a Japanese fishing vessel, the "Daigo Fukuryu Maru" (Lucky Dragon No. 5) suffered from exposure to radiation from the test. Even after the incident the US did not stop nuclear tests which continued to radioactively contaminate fish and rains in Japan. As a result, the petition movement for the ban of nuclear trials suddenly spread all over the country. By the summer of 1955 the number of the signatures grew to more than one third of Japan's population at the time. Under the strong influence of anti-nuclear Japanese public opinion the Science Council of Japan announced the so-called three principles of atomic energy: "openness," "democracy," and "independence" to ensure atomic energy was used for peaceful uses only. These principles were included in the Atomic Energy Basic Law established in December 1955. With this law, military uses of nuclear energy were strictly forbidden.

  18. Screening for celiac disease in Down's syndrome patients revealed cases of subtotal villous atrophy without typical for celiac disease HLA-DQ and tissue transglutaminase antibodies

    Institute of Scientific and Technical Information of China (English)

    Oivi Uibo; Kaupo Teesalu; Kaja Metsküla; Tiia Reimand; Riste Saat; Tarvo Sillat; Koit Reimand; Tiina Talvik; Raivo Uibo


    AIM: To investigate the prevalence of celiac disease (CD) as well as CD marker antibodies and susceptibility HLA-DQ haplotypes in 134 karyotyped Down's syndrome (DS) patients.METHODS: Immunoglobulin A (IgA) and G (IgG)type anti-gliadin antibodies (AGA), IgA type anti-tissue transglutaminase (tTG) antibodies (anti-tTG) with antigen of guinea pig and human source were determined by enzyme-linked immunosorbent assay and endomysium antibodies (EMA) by indirect immunofluoresence test.HLA-DQA1*0501/DQB1*0201 (DQ2) was revealed by polymerase chain reaction. Celiac disease was diagnosed by revised ESPGHAN criteria.RESULTS: 41% of DS patients had AGA, 6.0% IgAanti-tTG with guinea pig antigen, and 3.0 % IgA EMA (all positive for anti-tTG with human tTG). Subtotal villous atrophy was found in 5 out of 9 DS patients who had agreed to small bowel biopsy. One of them had DQA1*0501/DQB1*0201 and anti-tTG and EMA i.e. typical for CD markers (this case also fulfilled the ESPGHAN diagnostic criteria), but other four lacked these markers. Three non-biopsied DS patients had also most probably CD because DQA1*0501/DQB1*0201 and IgA anti-tTG (EMA) were detected. Thus, the prevalence of CD among our DS patients population is 3.0 % (95 %of confidence interval [CI]: 0.1-5.9 %).CONCLUSION: We confirm the increased frequency of CD among DS patients. In addition, we have revealed a subgroup of patients with subtotal villous atrophy but without characteristic for CD immunological and genetic markers. Whether these cases represent CD (with atypical immunopathogenesis) or some other immune enteropathy, requires further investigations.

  19. Screening and identification of T helper 1 and linear immunodominant antibody-binding epitopes in spike 1 domain and membrane protein of feline infectious peritonitis virus. (United States)

    Takano, Tomomi; Morioka, Hiroyuki; Gomi, Kohji; Tomizawa, Keisuke; Doki, Tomoyoshi; Hohdatsu, Tsutomu


    Feline infectious peritonitis virus (FIP virus: FIPV) causes a fatal disease in wild and domestic cats. The development of an FIP-preventive vaccine requires an antigen that does not induce antibody-dependent enhancement, and T helper (Th)1 activity plays an important role in protect against FIPV infection. In the present study, we identified synthetic peptides including Th1 and a linear immunodominant antibody-binding epitope in the S1 domain and M protein of FIPV. We also identified peptides that strongly induce Th1 activity from those derived from the structural proteins (S, M, and N proteins) of FIPV based on this and previous studies (Satoh et al. [19]). No Th1 epitope-containing peptide was identified in the peptides derived from the S1 domain of type I FIPV. In contrast, 7 Th1 epitope-containing peptides were identified in the S1 domain of type II FIPV, and no linear immunodominant antibody-binding epitope was contained in any of these peptides. Eleven Th1 epitope-containing peptides common to each serotype were identified in the M protein-derived peptides, and 2 peptides (M-11 and M-12) contained the linear immunodominant antibody-binding epitope. Of the peptides derived from the S, M, and N proteins of FIPV, those that induced significantly stronger Th1 activity than that of the FIPV antigen were rescreened, and 4 peptides were identified. When 3 of these peptides (M-9, I-S2-15, and II-S1-24) were selected and administered with CpG-ODNs to SPF cats, M-9 and II-S1-24 induced Th1 activity. Our results may provide important information for the development of a peptide-based vaccine against FIPV infection.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Evaluation of an indirect enzyme-linked immunosorbent assay for routine screening of Theiler's murine encephalomyelitis virus antibodies in mice colonies. (United States)

    Laborde, Juan M; Carbone, Cecilia; Corva, Santiago G; Galosi, Cecilia M


    The current study demonstrates the ability of an indirect enzyme-linked immunosorbent assay (iELISA) to detect antibodies against Theiler's murine encephalomyelitis virus in mice colonies. The antigen was produced from infected baby hamster kidney (BHK)-21 cells and treated with 1% Nonidet P40 in saline buffer. Control antigen was prepared following the same procedure using uninfected BHK-21 cells. The optimal antigen and serum dilutions were established. The reaction was revealed using an anti-mouse-horseradish peroxidase conjugate and 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). Optimized iELISA was validated by detection of antibodies in known positive and negative serum samples before testing the samples of unknown status. Performance of the iELISA was compared with the indirect fluorescent antibody test, and the cutoff value was determined by receiver operating curve. Indirect ELISA showed 100% sensitivity, 99.38% specificity, and 97.78% predictive positive value. The antigen used is easy to produce, and no special equipment is required. The iELISA developed is simple and provides a rapid and less costly tool for diagnosis and research.

  2. Visualization of high-throughput and label-free antibody-polypeptide binding for drug screening based on microarrays and surface plasmon resonance imaging (United States)

    Chen, Shengyi; Deng, Tao; Wang, Tongzhou; Wang, Jia; Li, Xin; Li, Qiang; Huang, Guoliang


    This work presents a visualization method for the high-throughput monitoring of antibody-polypeptide binding by integrating a microarray chip with surface plasmon resonance imaging (SPRi). A prism-coupled SPRi system with smart images processing software and a 5×5 polypeptide microarray was developed. The modeling analysis was performed to optimize the system and the materials of prism and chip, looking for the optimal incident wavelength and angle of incidence for dynamic SPRi detection in solution. The system can dynamically monitor 25 tunnels of biomolecule interactions in solution without secondary tag reactants. In addition, this system can determine the specific profile of antibody-polypeptide binding in each tunnel and yield a visual three-dimensional histogram of dynamic combinations in all microarray tunnels. Furthermore, the detection limit of the label-free antibody-polypeptide binding reached 1 pg/μL in a one-step binding test, and an ultrasensitive detection of 10 fg/μL was obtained using three-step cascade binding. Using the peptide microarray, the amount of sample and reagents used was reduced to 80 nL per tunnel, and 20×20 tunnels of biomolecule interactions could be analyzed in parallel in a 7 mm×7 mm microreaction cells. This device and method offer a potential platform for high-throughput and label-free dynamic monitoring multiple biomolecule interactions for drug discovery and basic biomedical research.

  3. Engineering antibodies by yeast display. (United States)

    Boder, Eric T; Raeeszadeh-Sarmazdeh, Maryam; Price, J Vincent


    Since its first application to antibody engineering 15 years ago, yeast display technology has been developed into a highly potent tool for both affinity maturing lead molecules and isolating novel antibodies and antibody-like species. Robust approaches to the creation of diversity, construction of yeast libraries, and library screening or selection have been elaborated, improving the quality of engineered molecules and certainty of success in an antibody engineering campaign and positioning yeast display as one of the premier antibody engineering technologies currently in use. Here, we summarize the history of antibody engineering by yeast surface display, approaches used in its application, and a number of examples highlighting the utility of this method for antibody engineering.

  4. Screening and identification of T helper 1 and linear immunodominant antibody-binding epitopes in the spike 2 domain and the nucleocapsid protein of feline infectious peritonitis virus. (United States)

    Satoh, Ryoichi; Furukawa, Tomoko; Kotake, Masako; Takano, Tomomi; Motokawa, Kenji; Gemma, Tsuyoshi; Watanabe, Rie; Arai, Setsuo; Hohdatsu, Tsutomu


    The antibody-dependent enhancement (ADE) of feline infectious peritonitis virus (FIPV) infection has been recognized in experimentally infected cats, and cellular immunity is considered to play an important role in preventing the onset of feline infectious peritonitis (FIP). In the present study, we synthesized eighty-one kinds of peptides derived from the spike (S)2 domain of type I FIPV KU-2 strain, the S2 domain of type II FIPV 79-1146 strain, and the nucleocapcid (N) protein of FIPV KU-2 strain. To detect the T helper (Th)1 epitope, peripheral blood mononuclear cells (PBMCs) obtained from FIPV-infected cats were cultured with each peptide, and Th1-type immune responses were measured using feline interferon (fIFN)-γ production as an index. To detect the linear immunodominant antibody-binding epitope, we investigated the reactivity of plasma collected from FIPV-infected cats against each peptide by ELISA. Four and 2 peptides containing Th1 epitopes were identified in the heptad repeat (HR)1 and inter-helical (IH) regions of the S2 domain of type I FIPV, respectively, and these were located on the N-terminal side of the regions. In the S2 domain of type II FIPV, 2, 3, and 2 peptides containing Th1 epitopes were identified in the HR1, IH, and HR2 regions, respectively, and these were mainly located on the C-terminal side of the regions. In the S2 domain of type I FIPV, 3 and 7 peptides containing linear immunodominant antibody-binding epitopes were identified in the IH and HR2 regions, respectively. In the S2 domain of type II FIPV, 4 peptides containing linear immunodominant antibody-binding epitopes were identified in the HR2 region. The Th1 epitopes in the S2 domain of type I and II FIPV were located in different regions, but the linear immunodominant antibody-binding epitopes were mostly located in the HR2 region. Eight peptides containing Th1 epitopes were identified in N protein, and 3 peptides derived from residues 81 to 100 and 137 to 164 showed strong

  5. Antiphospholipid antibody syndrome presenting with hemichorea. (United States)

    Ayalew, Yezenash; Khattak, Fazlihakim


    A 25-year-old Bangladeshi lady presented to neurology with a three-month history of involuntary movements of her right arm, associated with loss of power. There was progression to the right leg, and she subsequently developed episodes of slurred speech and blurred vision. At the time of presentation, she was 12 weeks pregnant and the symptoms were reported to have started at conception. Past medical history was unremarkable apart from one first trimester miscarriage and there was no significant family history suggestive of a hereditary neurological condition. MRI of the head revealed no abnormalities but serology showed positive antinuclear antibodies (ANAs) at a titre of 1/400. Further investigations revealed strongly positive anticardiolipin antibodies (>120) and positive lupus anticoagulant antibodies. The patient had a second miscarriage at 19 weeks gestation strengthening the possibility that the chorea was related to antiphospholipid antibody syndrome and she was started on a reducing dose of Prednisolone 40 mg daily and aspirin 300 mg daily. Six months later, she had complete resolution of neurological symptoms. There are several reports of chorea as a feature of antiphospholipid syndrome, but no clear consensus on underlying pathophysiology.

  6. Antiphospholipid Antibody Syndrome Presenting with Hemichorea

    Directory of Open Access Journals (Sweden)

    Yezenash Ayalew


    Full Text Available A 25-year-old Bangladeshi lady presented to neurology with a three-month history of involuntary movements of her right arm, associated with loss of power. There was progression to the right leg, and she subsequently developed episodes of slurred speech and blurred vision. At the time of presentation, she was 12 weeks pregnant and the symptoms were reported to have started at conception. Past medical history was unremarkable apart from one first trimester miscarriage and there was no significant family history suggestive of a hereditary neurological condition. MRI of the head revealed no abnormalities but serology showed positive antinuclear antibodies (ANAs at a titre of 1/400. Further investigations revealed strongly positive anticardiolipin antibodies (>120 and positive lupus anticoagulant antibodies. The patient had a second miscarriage at 19 weeks gestation strengthening the possibility that the chorea was related to antiphospholipid antibody syndrome and she was started on a reducing dose of Prednisolone 40 mg daily and aspirin 300 mg daily. Six months later, she had complete resolution of neurological symptoms. There are several reports of chorea as a feature of antiphospholipid syndrome, but no clear consensus on underlying pathophysiology.

  7. CMIA法测定梅毒螺旋体抗体在临床筛检梅毒的应用%Evaluation of a chemiluminescent microparticle immunoassay for determination of Treponema pallidum antibodies in syphilis screening test

    Institute of Scientific and Technical Information of China (English)

    金国江; 邹大伟; 杜毅鑫; 陈俊丽


    目的:探讨CMIA法测定梅毒螺旋体抗体在临床筛检梅毒的应用.方法:用CMIA法测定患者梅毒螺旋体抗体,阳性标本再进行TPPA确认试验,用ROC曲线分析确定S/CO值筛检梅毒精确度.结果:在28468例受检患者中,CMIA法梅毒抗体阳性501例,其中TPPA阳性413例,通过ROC曲线分析确定S/CO为3.2时(曲线下面积0.953,95%可信区间0.936~0.970),预测梅毒精确度为(敏感性88.8%,特异性94.4%).结论:CMIA法检测梅毒抗体可以准确预测梅毒感染,根据S/CO值可以确定是否需要进行TPPA确认试验.%Objective: To explore the clinical application of a chemiluminescent microparticle immunoassay (CMIA) for determination of Treponema pallidum antibodies in syphilis screening test. Methods: All serum samples were detected for the Treponema pallidum antibodies by CMIA. Then the positive samples were confirmed by Treponema pallidum particle agglutination ( TPPA). Receiver - operating characteristic ( ROC ) curve analysis was performed to evaluate the diagnostic accuracy of Treponema pallidum antibody S/CO for syphilis diagnosis. Results: In total of 28,468 patients, 501 patients were CMIA positive, and 413 cases were TPPA positive. By ROC curve a-nalysis, Treponema pallidum antibody S/CO ratio ( area, 0. 953 ; 95% confidence interval, 0. 936 to 0. 970) accurately predicted the presence of Syphilis, with a cutoff value of 3. 2 ( sensitivity, 88. 8% ; specificity, 94. 4% ) . Conclusion: The CMIA for determination of Treponema pallidum antibodies can predict syphilis infection accurately. The S/CO ratio can be used to determine whether a TPPA test should be performed.

  8. Update: Routine screening for antibodies to human immunodeficiency virus, civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components, January 2011-June 2016. (United States)


    This report contains an update through June 2016 of the results of routine screening for antibodies to the human immunodeficiency virus (HIV) among civilian applicants for military service and among members of the active and reserve components of the U.S. Armed Forces. During the surveillance period, annual seroprevalences among civilian applicants for military service peaked in 2015 (0.31 per 1,000 tested), up 29% from 2014 (0.24 per 1,000 tested). Seroprevalences among Marine Corps reservists, Navy active component service members, and Navy reservists also peaked in 2015. In the Army National Guard and the reserve component of the Marine Corps, full-year seroprevalences have trended upward since 2011. Overall (January 2011-June 2016) seroprevalences were highest for Army reservists, Army National Guard members, Navy active component members, and Navy reservists. Among active and reserve component service members, seroprevalences continue to be higher among Army and Navy members and males than their respective counterparts.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. [Sensitivity of screening kits for anti-HIV antibodies. 1999 update. Retrovirus Working Group of the French Society for Blood Transfusion]. (United States)

    Couroucé, A M


    A comparative evaluation of the sensitivity of anti-HIV screening assays has been recently performed with a selected panel of 65 samples which included HIV1 group M (per-seroconversions, seroconversions and seropositives infected with genotypes A, B, C, D, E), HIV1 group O and HIV2. The results obtained with the 21 ELISA HIV1 + HIV2 screening assays are presented. Among these 21 assays, four are combined anti-HIV and p24 Ag assays. All the assays except four fulfilled the criteria defined at the beginning of the study, i.e., positive results on all the seropositives including seroconversions and positive results on at least 50% of the per-seroconversions. The main differences were observed with the ten per-seroconversion samples, the number of positive results on such samples varying from three to ten. The constant improvement of anti-HIV screening tests which leads one to shorten the 'window' period permits and earlier diagnosis of HIV infection and a progressive decrease of the transfusional risk.

  11. Hepatitis B virus screening in contacts of blood donors with antibodies against core protein (anti-HBc, but without surface antigen (HBsAg

    Directory of Open Access Journals (Sweden)

    Hildenete Monteiro Fortes


    Full Text Available To increase blood safety Brazil introduced screening for anti-HBc among blood donors in 1993. There was a decrease in the hepatitis B virus (HBV transmission, but this measure identified a great number of HBsAg-negative, anti-HBc-positive donors. Surveillance policy determines that contacts of HBV carriers should be screened to HBV markers, but there is no recommendation about how to guide contacts of HBsAg-negative, anti-HBc-positive donors. Aiming to evaluate whether the contacts of this group are at greater risk for HBV infection, a cross-sectional study was performed to compare prevalence of HBV infection between contacts of HBsAg-positive blood donors (group I and contacts of HBsAg-negative, anti-HBc-positive donors (group II. Contacts were submitted to a questionnaire and blood tests for HBV markers. In group I (n = 143, 53 (37.1% were anti-HBc-positive and 11 (7.7% were HBsAg-positive. In group II (n = 111, there were 9 and 0.9%, respectively. HBV exposure was associated with group I, sexual activity, blood transfusion, being one of the donor's parents, and living for more than ten years with the donor. Regarding the families as sample units, it was more common to find at least one member with HBV markers (p < 0.05 among the families of group I compared to group II. Contacts of HBsAg-negative, anti-HBc-positive individuals presented a much lower risk of having already been exposed to HBV and there is no need to screen them for HBV in low to moderate prevalence populations.

  12. Updates: Routine screening for antibodies to HIV-1, civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components. (United States)


    During routine testing of civilian applicants for U.S. military service, the overall seroprevalence of antibodies to HIV-1 was lower in 2010 than in any year since 1990. Among members of the active components of the U.S. Army and Air Force, HIV-1 seroprevalences were higher in 2008-2010 than in recent prior years. Among members of the active components of the U.S. Navy and Marine Corps, the Marine Corps Reserve, and the Army National Guard, HIV-1 seroprevalences have slightly declined or remained relatively stable for at least ten years. In the reserve components of most of the service branches, it is difficult to discern long-term trends because of instability of seroprevalences observed in the relatively small numbers of reserve component members tested each year.

  13. Antibody-based screening of cell wall matrix glycans in ferns reveals taxon, tissue and cell-type specific distribution patterns

    DEFF Research Database (Denmark)

    Leroux, Olivier; Sørensen, Iben; Marcus, Susan E.;


    plants, ferns have been largely neglected in cell wall comparative studies. Results: To explore fern cell wall diversity sets of monoclonal antibodies directed to matrix glycans of angiosperm cell walls have been used in glycan microarray and in situ analyses with 76 fern species and four species...... across the ferns and specifically associated with phloem cell walls and similarly the LM11 xylan epitope was associated with xylem cell walls. The LM5 galactan and LM6 arabinan epitopes, linked to pectic supramolecules in angiosperms, were associated with vascular structures with only limited detection...... in ground tissues. Mannan epitopes were found to be associated with the development of mechanical tissues. We provided the first evidence for the presence of MLG in leptosporangiate ferns. Conclusions: The data sets indicate that cell wall diversity in land plants is multifaceted and that matrix glycan...

  14. Role of signal-to-cut-off ratios of anti-hepatitis C virus antibody by enzyme immunoassays along with ID-NAT for screening of whole blood donors in India

    Directory of Open Access Journals (Sweden)

    Satyam Arora


    Full Text Available Background: The use of elevated signal-to-cut off ratios (S/CO as an alternate to further supplemental testing (i.e., RIBA has been included in the guidelines provided by the Centres for Disease Control and Prevention for HCV diagnostic purposes since 2003. With availability of screening by NAT and non availability of RIBA, further confirmation of HCV infection has been possible at the molecular level (RNA. Aims: To study the role of S/CO ratios of anti hepatitis C virus antibody detection by enzyme immunoassays (EIA along with ID-NAT for screening of whole blood donors. Methods: In this study we reviewed the donor screening status for anti HCV from January 2013 to May 2014. All the donations were screened for anti HCV with fourth generation ELISA (BioRad Monolisa Ag-Ab Ultra as well as with ID NAT (Procleix Ultrio. The S/CO ratio of all the anti-HCV reactive samples were analysed for their presence of HCV RNA. Results: On screening 21,115 donors for HCV, 83 donors (0.39% were found reactive on pilot tube and repeat plasma bag testing (S/Co ratio ≥1 by ELISA. 41 donors were HCV RNA reactive with ID-NAT. 4 samples out of 41 were NAT yields and 37 were concordant reactive with ELISA. The S/Co ratio of anti-HCV reactive samples ranged from 0.9-11.1 [mean = 5.1; SD ΁ 2.9] whereas S/Co ratio of anti HCV and NAT reactive samples (concordant positives ranged from 4.1-11.1 [mean 7.3]. In our analysis we found that S/CO ratio of 4 showed positive predictive value (PPV and sensitivity of 100%. Summary/Conclusions: Our study showed that S/CO of 4 for anti HCV on ELISA would have maximum positive predictive value of having donor with HCV RNA. S/CO ratio of 4 is very close to 3.8 which was the CDC guideline. The presence of anti-HCV does not distinguish between current or past infections but a confirmed anti-HCV-positive result indicates the need for counseling and medical evaluation for HCV infection.

  15. The application of cassette automatic blood analyzer in irregular antibody screening for blood donors%卡式全自动血型仪在献血者不规则抗体筛查中的应用

    Institute of Scientific and Technical Information of China (English)

    邹姣丽; 王庆; 邹文涛; 何子毅; 王若珩; 陈庆恺; 崔四平


    目的:比较卡式全自动血型仪法与传统血清学法对不规则抗体筛查的应用效果。方法分别采用卡式全自动血型仪法与传统血清学方法对2013年10月1日至2014年4月30日东莞市无偿献血者45709份标本进行不规则抗体筛查,对筛查阳性的标本采用抗球蛋白法进行鉴定,比较两种方法检出不规则抗体的性能,并对献血者不规则抗体分布进行分析。结果卡式全自动血型仪检出不规则抗体阳性95例,检出率为0.208%,传统血清学法检出不规则抗体16例,检出率0.035%,差异有统计学意义( P<0.05);卡式全自动血型仪法阳性检出符合率为56.547%,高于传统血清学法阳性检出符合率28.571%( P<0.05);RhD阴性献血者不规则抗体阳性率为2.242%,高于RhD阳性献血者不规则抗体率0.198%(P<0.05)。结论卡式全自动血型仪法鉴定献血者红细胞不规则抗体的阳性检出率高于传统血清学法,对RhD阴性献血者更应进行不规则抗体的筛查,献血者红细胞不规则抗体类型以Ig M型无临床意义的抗体为主。%Objective To compare the application effect between the method of cassette automatic blood analyzer and the method of traditional serological .Methods Using the method of cassette automatic blood analyzer and the method of traditional serological , respectively ,the samples of blood donors in Dongguan from October 1 ,2013 to April 30 ,2014 were performed irregular antibody screening .The positive samples of screening were identified by using antiglobulin method ,and the irregular antibodies of blood do‐nors were analyzed .Results There were 95 positive cases of irregular antibody by the method of cassette automatic blood analyzer , and the detection rate was 0 .208% .There were 16 positive cases of irregular antibody by the method of traditional serological ,the detection rate was 0 .035% ,and there was

  16. Effect of using heat-inactivated specimens with several HIV antibody screening and confirmatory assay kits%样品热灭活对HIV抗体筛查和确证试验结果的影响研究

    Institute of Scientific and Technical Information of China (English)

    郎文文; 张桂云; 邱景富; 曹薇; 李华荣; 邱茂锋; 蒋岩


    目的 研究血浆样品经热灭活(56℃30min)后,其艾滋病病毒(HIV)抗体筛查和确证试验结果是否会受到影响.方法 取5份HIV抗体阳性血浆样品,各分为2管,其中1管进行热灭活处理,然后对这2组样品分别进行10倍系列稀释,用1种HIV抗体酶联免疫吸附试验(ELISA)试剂检测,比较S/CO比值.取300份血浆样品,分为灭活组和普通组,分别用5种HIV抗体ELISA试剂(其中第三代试剂3种、第四代试剂2种)和3种HIV抗体快速检测试剂进行筛查检测,出现筛查阳性反应的样品进一步做确证试验.结果 随着稀释度的增加,5份HIV抗体阳性样品灭活前后的ELISA检测结果(S/CO比值)都逐渐减小,直至转为阴性反应,其中4份样品灭活后比灭活前早一个稀释度转阴,1份同时转阴,提示灭活过程会略微降低HIV抗体的浓度.用5种ELISA试剂、3种快速检测试剂检测300份样品,样品灭活前后的检测结果差异均无统计学意义;对出现筛查阳性反应的所有样品,灭活前后的确证试验检测结果一致.结论 经56℃30min热灭活后,尽管HIV阳性血浆样品中的HIV抗体浓度略有降低,但对于未经稀释的常规临床血浆样品来说,热灭活处理不会明显影响目前常用筛查、确证试剂的检测结果.%Objective To study the effect of using heat-inactivated (56 ℃ , 30min) plasma specimens with several HIV antibody screening and confirmatory assay kits. Methods Five HIV antibody-positive plasma specimens were aliquoted into 2 vials, respectively. One group of them was heat-inactivated by 56 ℃, 30min. Both groups were 1 : 10 serially diluted and then detected by an ELISA kit for HIV antibody. Three hundred plasma specimens were aliquoted into 2 vials, respectively, and one group of them was heat-inactivated. Both groups were detected by 5 ELISA kits (including 3 HIV antibody and 2 HIV antigen/antibody ELISAs) and 3 HIV rapid tests, respectively. The repeat-reactive specimens

  17. 7251例不规则抗体筛查结果分析%The Analysis of 7251 Cases Irregular Anti-bodies Screening

    Institute of Scientific and Technical Information of China (English)

    陈志远; 尹婷婷


    Objective Detected 7251 cases of irregular antibodies ( Irregular antibody,IA) type,then analysis potency fac-tors of IA positive producing and assessed clinical adverse events related IA positive. Methods Used salt water,micro gel method for prenatal immunological examination to determine qualitative and quantitative of IA,United clinical data were retrospectively analyzed IA impact on clinical adverse events. Results In 7251 cases of patients,IA-positive 54 cases(0. 61%),of which 31 cases(0. 42%) Rh system,including anti-D 23cases,anti-E 4 cases,anti-C2 cases,anti-c 1case,anti-e 1 case;other blood group system identified 8 cases(0. 19%),of which the anti-M3 cases,anti-Mur 2 cases,anti-Lea2 cases,anti-JKb1 cases;auto-antibodies detected in six cases;specific antibodies were not detected in 10 cases. The RA was higher occurrence ration in patients with antigen AB and Rh-,and the IA patented clinical value when antibody titer≥128. In IA-positive patients in 54 cases,12 cases of hepatitis patients(12/1500, 1. 47%),for 27 cases of pregnant women(27/2028,1. 33%),autoimmune diseases six cases(6/477,1. 26%),cancer patients five cases(5/2523,0. 20%),other four cases(4/723,0. 55%). This study further analyzed the correlation between IA and diseases,and found 45 cases of hemolytic reaction in IA-positive patients,that there were different degrees of jaundice and after clinical diagnosis confirmed;but patients with hemolytic reactions in 45 cases,the clinical inspection has 28 cases of patients with no history of blood transfusion and pregnancy history, but there were different degrees of inflammation. Conclusion The investigation included that IA played a role in clinic adverse events by according to hemolytic reaction,the relationship of IA and inflammation was discovered. The results of study may appear that the detection of IA had important value to decrease adverse transfusion reaction,at the same time,it may early warn the hemolytic reaction in persistent

  18. [Renal histology in 44 patients with specific antibodies of soluble nuclear antigens]. (United States)

    Meyer, O; Gaudreau, A; Peltier, A P


    The authors studied the correlations between renal histology and specific antinuclear antibodies of soluble nuclear antigens (anti-Sm, anti-RNP, anti-protein) in 44 patients with such auto-antibodies. They were mostly patients with lupus erythematosus (35/44), more rarely mixed collagen disease or Sjögren's disease. The presence of any one of the specific antibodies of nuclear antigens is not associated with any special renal prognosis; thus the presence of anti-RNP does not mean that there are no histological renal lesions. The renal prognosis depends in fact on the presence of anti-ADN native antibodies. Among the other laboratory parameters (rheumatoid factors, complement levels, cryoglobulinemia) only hypocomplementemia seems to be associated with a poor renal prognosis, the presence of rheumatoid factor has perhaps a protective role.

  19. Belgian recommendations on ANA, anti-dsDNA and anti-ENA antibody testing. (United States)

    Van Blerk, M; Bossuyt, X; Humbel, R; Mewis, A; Servais, G; Tomasi, J P; Van Campenhout, C; Van Hoovels, L; Vercammen, M; Damoiseaux, J; Coucke, W; Van de Walle, P


    Autoantibodies to nuclear antigens, i.e. antinuclear antibodies (ANA), antibodies to double-stranded DNA (dsDNA) and extractable nuclear antigens (ENA), are useful as diagnostic markers for a variety of autoimmune diseases. In March 2010, the Belgian national External Quality Assessment Scheme sent a questionnaire on ANA, anti-dsDNA and anti-ENA antibody testing designed by the Dutch EASI (European Autoimmunity Standardization Initiative) team, to all clinical laboratories performing ANA testing. Virtually all laboratories completed the questionnaire (97·7%, 127/130). This paper discusses the results of this questionnaire and provides valuable information on the state-of-the-art of ANA, anti-dsDNA and anti-ENA antibody testing as practiced in the Belgian laboratories. In addition, this work presents practical recommendations developed by the members of the advisory board of the scheme as a result of the outcome of this study.

  20. Identification of antigen-specific human monoclonal antibodies using high-throughput sequencing of the antibody repertoire. (United States)

    Liu, Ju; Li, Ruihua; Liu, Kun; Li, Liangliang; Zai, Xiaodong; Chi, Xiangyang; Fu, Ling; Xu, Junjie; Chen, Wei


    High-throughput sequencing of the antibody repertoire provides a large number of antibody variable region sequences that can be used to generate human monoclonal antibodies. However, current screening methods for identifying antigen-specific antibodies are inefficient. In the present study, we developed an antibody clone screening strategy based on clone dynamics and relative frequency, and used it to identify antigen-specific human monoclonal antibodies. Enzyme-linked immunosorbent assay showed that at least 52% of putative positive immunoglobulin heavy chains composed antigen-specific antibodies. Combining information on dynamics and relative frequency improved identification of positive clones and elimination of negative clones. and increase the credibility of putative positive clones. Therefore the screening strategy could simplify the subsequent experimental screening and may facilitate the generation of antigen-specific antibodies.

  1. 第四代HIV抗原抗体检测试剂在血液筛查中的应用%Application of 4th generation reagents for the detection of HIV antigen and antibody in blood screening

    Institute of Scientific and Technical Information of China (English)

    陈尚良; 郑欣; 曾月婷; 廖扬勋; 梁洁贞; 余文潮; 李结敏; 梁丽婷; 梁剑锋


    目的:研究第四代HIV抗原抗体检测试剂在血液筛查中的应用价值。方法对第四代HIV抗原抗体检测试剂进行灵敏度和重复性实验;分别采用第三代HIV抗体检测试剂和第四代HIV抗原抗体检测试剂对血液样本进行初复检,对第三代试剂检测阴性第四代试剂检测阳性的样本采用HIV核酸检测方法进行确认。结果第四代HIV抗原抗体检测试剂最低抗原检出浓度为1.25 U/mL,孔间精密度为3.2%;2009年10月~2011年3月,共筛查样本23480例,其中13例第三代试剂结果阴性而第四代试剂结果阳性,经HIV核酸检测的方法进行确认,全部结果均为阴性。结论在血液筛查中引入第四代HIV检测试剂的试剂功效尚待进一步验证,在已经广泛开展HIV抗体检测的基础上,引入第四代HIV抗原抗体检测试剂还是引入核酸检测具有更好的成本效益比,是一个值得待探讨的课题。%Objective To study the application of 4th-generation HIV reagents for detecting HIV antigens and antibodies in blood screening. Methods The sensitivity and repeatability experiments were tested respectively. All samples were detected by 3th-generation HIV reagents for only detecting HIV antibody and 4th-generation HIV reagents for detecting HIV antigens and antibodies, respectively, and the samples which were positive in 4th-generation HIV reagents detecting but negative in 3th-generation HIV reagents were confirmed by HIV nucleic acid detections. Results The minimum detectable concentration of HIV antigen was 1.25 U/mL and the precision was 3.2%for 4th-generation HIV reagent. In tolal 23 480 samples had been screened from October 2009 to March 2011, 13 samples were positive in 4th-generation HIV reagent detecting but negative in HIV 3th-generation HIV reagent, all of them were confired negative by HIV nucleic acid detection. Conclusion The efficacy of HIV detection attribute to applying 4th-generation HIV reagent

  2. Overcoming a "probable" diagnosis in antimitochondrial antibody negative primary biliary cirrhosis: study of 100 sera and review of the literature. (United States)

    Bizzaro, Nicola; Covini, Giovanni; Rosina, Floriano; Muratori, Paolo; Tonutti, Elio; Villalta, Danilo; Pesente, Fiorenza; Alessio, Maria Grazia; Tampoia, Marilina; Antico, Antonio; Platzgummer, Stefan; Porcelli, Brunetta; Terzuoli, Lucia; Liguori, Marco; Bassetti, Danila; Brusca, Ignazio; Almasio, Piero L; Tarantino, Giuseppe; Bonaguri, Chiara; Agostinis, Paolo; Bredi, Elena; Tozzoli, Renato; Invernizzi, Pietro; Selmi, Carlo


    Serum anti-mitochondrial antibodies (AMA) are the serological hallmark of primary biliary cirrhosis (PBC), yet up to 15% of PBC sera are AMA negative at routine indirect immunofluorescence (IIF) while being referred to as "probable" cases. The diagnostic role of PBC-specific antinuclear antibodies (ANA) remains to be determined. We will report herein data on the accuracy of new laboratory tools for AMA and PBC-specific ANA in a large series of PBC sera that were AMA-negative at IIF. We will also provide a discussion of the history and current status of AMA detection methods. We included IIF AMA-negative PBC sera (n=100) and sera from patients with other chronic liver diseases (n=104) that had been independently tested for IIF AMA and ANA; sera were blindly tested with an ELISA PBC screening test including two ANA (gp210, sp100) and a triple (pMIT3) AMA recombinant antigens. Among IIF AMA-negative sera, 43/100 (43%) manifested reactivity using the PBC screening test. The same test was positive for 6/104 (5.8%) control sera. IIF AMA-negative/PBC screen-positive sera reacted against pMIT3 (11/43), gp210 (8/43), Sp100 (17/43), both pMIT3 and gp210 (1/43), or both pMIT3 and Sp100 (6/43). Concordance rates between the ANA pattern on HEp-2 cells and specific Sp100 and gp210 ELISA results in AMA-negative subjects were 92% for nuclear dots and Sp100 and 99% for nuclear rim and gp210. Our data confirm the hypothesis that a substantial part of IIF AMA-negative (formerly coined "probable") PBC cases manifest disease-specific autoantibodies when tested using newly available tools and thus overcome the previously suggested diagnostic classification. As suggested by the recent literature, we are convinced that the proportion of AMA-negative PBC cases will be significantly minimized by the use of new laboratory methods and recombinant antigens.

  3. In vitro screening of major neurotransmitter systems possibly involved in the mechanism of action of antibodies to S100 protein in released-active form

    Directory of Open Access Journals (Sweden)

    Gorbunov EA


    Full Text Available Evgeniy A Gorbunov, Irina A Ertuzun, Evgeniya V Kachaeva, Sergey A Tarasov, Oleg I EpsteinOOO “NPF “MATERIA MEDICA HOLDING”, Moscow, Russian FederationAbstract: Experimentally and clinically, it was shown that released-active form of antibodies to S100 protein (RAF of Abs to S100 exerts a wide range of pharmacological activities: anxiolytic, antiasthenic, antiaggressive, stress-protective, antihypoxic, antiischemic, neuroprotective, and nootropic. The purpose of this study was to determine the influence of RAF of Abs to S100 on major neurotransmitter systems (serotoninergic, GABAergic, dopaminergic, and on sigma receptors as well which are possibly involved in its mechanism of pharmacological activity. Radioligand binding assays were used for assessment of the drug influence on ligand–receptor interaction. [35S]GTPγS binding assay, cyclic adenosine monophosphate HTRF™, cellular dielectric spectroscopy assays, and assays based on measurement of intracellular concentration of Ca2+ ions were used for assessment of agonist or antagonist properties of the drug toward receptors. RAF of Abs to S100 increased radioligand binding to 5-HT1F, 5-HT2B, 5-HT2Cedited, 5-HT3, and to D3 receptors by 142.0%, 131.9%, 149.3%, 120.7%, and 126.3%, respectively. Also, the drug significantly inhibited specific binding of radioligands to GABAB1A/B2 receptors by 25.8%, and to both native and recombinant human sigma1 receptors by 75.3% and 40.32%, respectively. In the functional assays, it was shown that the drug exerted antagonism at 5-HT1B, D3, and GABAB1A/B2 receptors inhibiting agonist-induced responses by 23.24%, 32.76%, and 30.2%, respectively. On the contrary, the drug exerted an agonist effect at 5-HT1A receptors enhancing receptor functional activity by 28.0%. The pharmacological profiling of RAF of Abs to S100 among 27 receptor provides evidence for drug-related modification of major neurotransmitter systems.Keywords: dopamine agent, released

  4. The Political Styles of Local Anti-Nuclear Waste Movements in Finland

    Energy Technology Data Exchange (ETDEWEB)

    Kojo, Matti [Univ. of Tampere (Finland). Dept. of Political Science and International Relations


    This paper aims to analyse the political styles of local anti-nuclear waste movements in Finland. The main focus is on the tension between the environmental impact assessment process (EIA) and the activity of local opposing groups. According to the EIA Act the purpose of the EIA process is to provide information and opportunities for citizens to participate in planning. It therefore aims to enhance the transparency of the decision-making process. The Finnish nuclear waste company, Posiva Oy, made great efforts locally to create opportunities for participation, but was much criticised by local activists. My questions are the following: How did these local movements participate in Posiva's EIA process? What kind of local differences were there in participating and how can these differences be explained? And finally; what did the EIA process mean to the political styles of movements? EIA meant a temporary change in political style on local level. Firstly; because it brought science into local politics very clearly. Secondly EIA with the Nuclear Energy Act framed decision-making with a timetable, which Posiva emphasised when local groups wanted to make a decision immediately. Thirdly EIA enabled the problem to be defined according to local needs but on the other hand plan level EIA separated nuclear waste issue from the use of nuclear power. Fourthly dialogue in the EIA process favoured rational discourse. Although local views were heard, the process did try to teach the people how to speak with the decision-making system, in the system's language. Thus the purpose of EIA was to make the local discussion controllable. All the local groups, the Romuvaara, Kivetty and Loviisa Movements and the Friends of the Earth in Pori region, took part in EIA dialogue but the reactions were different. The Romuvaara Movement was very active in participating and succeeded in exploiting EIA locally whereas the Loviisa Movement tried to displace the whole process. The Loviisa

  5. Update: routine screening for antibodies to human immunodeficiency virus, civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components, January 2010-June 2015. (United States)


    This report contains an update through June 2015 of the results of routine screening for antibodies to the human immunodeficiency virus (HIV) among civilian applicants for military service and among members of the active and reserve components of the U.S. Armed Forces. Seroprevalences among civilian applicants in 2014 and the first half of 2015 (0.21 and 0.22 per 1,000 tested, respectively) were markedly lower than in 2012 (0.28 per 1,000 tested). In nearly every component of every military service, seroprevalences in 2014 and 2015 were either lower than, or relatively similar to, prevalences in prior years; however, in the Army National Guard, seroprevalences increased each year and approximately doubled from 2010 (0.18 per 1,000 tested) to 2014-2015 (0.36-0.39 per 1,000 tested). Among active and reserve component service members, seroprevalences continue to be higher among Army and Navy members and males than their respective counterparts.

  6. Update: Routine screening for antibodies to human immunodeficiency virus, civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components, January 2009-June 2014. (United States)


    This report contains an update through June 2014 of the results of routine screening for antibodies to the human immunodeficiency virus (HIV) among civilian applicants for military service and among members of the active and reserve components of the U.S. Armed Forces. Seroprevalences among civilian applicants in 2013 and the first half of 2014 (0.19 and 0.15 per 1,000 tested, respectively) were markedly lower than in 2012 (0.27 per 1,000 tested). In nearly every component of every service, seroprevalences in 2013 and 2014 were either similar or lower than in prior years; however, in the Army National Guard, seroprevalences increased each year and approximately doubled from 2010 (0.18 per 1,000 tested) to 2013-2014 (0.35-0.41 per 1,000 tested). Among active and reserve component service members, seroprevalences continue to be higher among Army and Navy members and males than their respective counterparts.

  7. 孕产妇HIV抗体筛查阳性与免疫印迹试验对比%Comparison of Positive of HIV Antibody Screening and Western Blot Test in Pregnant Women

    Institute of Scientific and Technical Information of China (English)

    朱厚宏; 王晨笛; 刘杨; 张祁; 孟建彤; 黄薇


    Objective To explore the relationship between the results of HIV antibody screening test and Westem Blot test ( WB) in pregnant women. Methods Thirty three positive samples from pregnant women were tested with the ELISA and Westem blot test (WB). Those with indefinite result by WB test were re-tested 3 and 6 months later, and HIV-1 viral load testing ( VL) was carried out at 6th month. The results of ELISA and the WB test were analyzed. Result The 33 samples were all positive by ELISA and 20 were HIV-1 positive by WB test. The ELISA and the WB test positive coincidence rate was 60. 61 % (20/33). Of 20 samples 0were with S/CO value more than 8, the WB test results of HIV-1 antibody were all positive. Of 13 samples were with S/CO value less than 1 , 5 were negative and 8 were indefinite in WB test. The HIV-1 viral load test result at 6th month was less than the minimum detection limit (50 IU/ml ) . Conclusion The false positive in HIV antibody screening test (ELISA) was observed. The false positive sample were in the WB test negative and indefinite ones. ELISA and the WB test positive coincidence rate increased with the S/CO value of ELISA. The HIV WB test indefinite result in pregnant women was mostly due to non-specific immune response.%目的 探讨孕产妇人类免疫缺陷病毒(HIV)抗体筛查试验阳性与免疫印迹试验(WB)结果的关系.方法 对33份孕产妇HIV抗体待复查样本,进行酶联免疫吸附试验(ELISA )及免疫印迹试验(WB),对WB试验结果不确定者进行3个月、6个月随访检测,6个月随访样本补充HIV-1病毒载量检测,分析ELISA检测与WB试验检测结果.结果 33份样本ELISA复检均呈阳性,免疫印迹试验(WB)中20份确认为HIV-1抗体阳性,ELISA检测与WB试验阳性符合率为60.61%(20/33); S/CO值>8的20份样本,WB试验结果均为HIV-1抗体阳性,I < S/CO值<8的13份样本,WB试验5份为HIV抗体阴性,8份为HIV抗体不确定;6个月随访样本HIV-1病毒载量检测,

  8. 全人源抗鼻咽癌噬菌体单链抗体的筛选与鉴定%Screening and characterization of human scFv antibodies against nasopharyngeal carcinoma

    Institute of Scientific and Technical Information of China (English)

    李艳东; 谢平丽; 王甲甲; 李跃辉; 胡锦跃; 李官成


    目的 从全人源抗鼻咽癌噬菌体抗体库中筛选特异性单链抗体(ScFv),并对其特异性进行鉴定.方法 通过噬菌体表面展示技术把ScFv表达在噬菌体表面,以鼻咽癌细胞作为抗原,用抗原递减法,通过"吸附-洗脱-扩增"过程筛选并富集特异性抗体,及ELISA筛选,获得特异阳性克隆进行免疫组化鉴定并测序.结果 通过对抗体库进行三轮正负淘洗和富集后,随机挑选4212个克隆进行ELISA,发现3个克隆对CNE2呈强阳性反应,而与人正常细胞系HUVEC等呈弱阳性反应或不反应.对克隆HNSAO33进一步进行免疫细胞化学验证,结果与ELISA反应一致;免疫组织化学鉴定表明克隆HNSAO33与鼻咽癌组织和鼻咽组织阳性率的差别有统计学意义.结论 通过淘选富集、ELISA和免疫化学鉴定获得特异性较强的噬菌体克隆,为鼻咽癌发病机制的研究和临床诊断以及治疗奠定了基础.%Objective To screen the anti-nasopharyngeal carcinoma scFv from a human anti-nasopharyngeal carcinoma single-chain phage antibody library, and identify its characteristics. Methods The single-chain phage antibody library was subjected to three rounds of positive and negative cell panning and enrichment, and then it was selected by ELISA. The binding specificity of phage antibodies with naso-pharyngeal carcinoma cells was confirmed by immunohistochemistry. Results After panning, enrichment and testing by ELISA, 3 phage an-tibody clones reacting with CNE2 more strongly than HUVEC and NP69 were picked out from 4212 clones. One clone, HNSAO33, was fur-ther analyzed after DNA sequencing. The results of immunohistochemistry with cultured cells were similar to those of ELISA. HNSAO33 spe-cifically reacted to nasopharyngeal carcinoma cells in most human nasopharyngeal carcinoma tissue sections except a few human normal naso-pharyngeal tissue sections. The distinction of positive rates was of a great statistical significance. Conclusion ELISA

  9. 抗磷脂抗体对妊娠丢失筛查的临床价值%Clinical value of antiphospholipid antibody in the screening for pregnancy loss

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the relationship between history of pregnancy loss and antiphospholipid antibodies (APA),including anticardiolipin (ACA) and lupus anti-coagulant antibodies (LA).Methods One hundred and fifty patients with history of unexplained pregnancy loss as study group and 120 normal nonpregnant women as control group.The study group was further divided into three subgroups:embryo growth arrest (n =36),stillbirth (n =44),and recurrent abortion (n =70)levels of serum APA(including ACA and LA) were determined repectively and analyzed.Results The positive rates of APA,ACA,LA in the study group were significantly higher than those in control group (P < 0.05).The above significance was true in both stillbirth and recurrent abortion groups but not in embryo growth arrest group.Conclusions Levels of serum APA are associated with pregnancy loss,especially about recurrent abortion and stillbirth.We suggest routine screening of serum APA should be performed in patients with history of fetal wastage for the sake of early treatment.%目的 探讨妊娠丢失与抗磷脂抗体(APA)[包括抗心磷脂抗体(ACA)和狼疮抗凝抗体(LA)]的关系.方法 观察组为150例有妊娠丢失史的患者,其中分为胚胎停育组(36例)、死胎组(44例)和复发性流产组(70例).对照组为同期120例正常孕妇,分别测定观察组和对照组静脉血清APA水平,并进行对比分析.结果 整个观察组APA、ACA、LA的阳性率均高于对照组,差异有统计学意义(P<0.05).胚胎停育组与对照组比较,差异无统计学意义(P>0.05).死胎组和复发性流产组的APA、ACA、LA阳性率分别与对照组比较,差异均有统计学意义(P<0.05).结论 APA与妊娠丢失有关,尤其对于复发性流产和死胎者.因此,对有不良孕产史的患者常规筛查APA,有利于尽早对因治疗.

  10. Thyroid Antibodies (United States)

    ... e.g., at regular intervals after thyroid cancer treatment) Thyroid stimulating hormone receptor antibody, Thyroid Stimulating Immunoglobulin TRAb, TSHR Ab, TSI Graves disease When a person has symptoms of hyperthyroidism If a pregnant woman has a known autoimmune ...

  11. Serologic survey for antibodies to canine parvovirus and influenza virus in wild raccoon dogs (Nyctereutes procyonoides) in South Korea. (United States)

    Cha, Se-Yeoun; Seo, Hye-Suk; Kang, Min; Jang, Hyung-Kwan


    Sera from 102 wild raccoon dogs (Nyctereutes procyonoides) were screened for antibodies to canine parvovirus (CPV) and influenza A virus (IAV) in South Korea. Sixteen samples were antibody positive for CPV and all samples were negative for IAV antibodies.

  12. Analysis on positive confirmation result of HIV antibody prelimicnary screening in voluntary blood donation%无偿献血HIV抗体初筛阳性确认结果分析

    Institute of Scientific and Technical Information of China (English)

    李庚娣; 刘永梅


    目的:分析2009~2015年H IV 抗体筛查和确证试验结果,为制订在低危人群中招募献血者的招募策略和献血者回归提供依据。方法采用酶联免疫吸附试验(ELISA)检测 HIV抗体,分别用2个不同厂家试剂检测。HIV抗体检测阳性或可疑的标本送深圳市疾病预防控制中心进行免疫蛋白印迹确认。选取2009~2015年血液标本筛查确认结果进行统计分析。结果2009~2015年197766份血液标本,2009~2015年每年总的感染率分别为1.3/10万、4.7/10万、5.6/10万、5.4/10万、5.3/10万、4.6/10万、7.3/10万,平均感染率为0.0049%(4.9/10万),平均确诊阳性率为22.1%,假阳性率为77.9%。H IV抗体确认阳性的献血者中年龄18~30岁的占67.7%。结论重视献血前的征询招募工作,针对外来劳务工和文化程度相对低的献血者加强献血知识的普及,特别是有高危行为的献血者,采用小卡片的方式,引导其到疾控中心做专业的咨询检测,既提高输血安全系数又能预防艾滋病扩散。针对假阳性的献血者做好回访跟踪监测及献血资格的回归工作,减少献血者的流失。选择更加灵敏的试剂及选择更加灵敏的病毒核酸扩增的方法,可尽量缩小因窗口期感染的风险,确保输血安全。%Objective To analyze the screening and confirmation test results of human immunodeficiency virus (HIV) antibody during 2009-2015 to provide the basis for the recruiting strategies in recruiting blood donors among the low risk population and blood donor′s returning to the team .Methods The blood samples of voluntary blood donors were detected the HIV antibody by the ELISA method ,with the reagents provided by 2 different manufacturers .The samples of HIV antibody positive or suspected sam‐ples were submitted to the Shenzhen Municipal Center for Disease Control and Prevention CDC for conducting

  13. Antiparietal cell antibody test (United States)

    APCA; Anti-gastric parietal cell antibody; Atrophic gastritis - anti-gastric parietal cell antibody; Gastric ulcer - anti-gastric parietal cell antibody; Pernicious anemia - anti-gastric parietal cell antibody; ...

  14. Female Infertility and Serum Auto-antibodies: a Systematic Review. (United States)

    Deroux, Alban; Dumestre-Perard, Chantal; Dunand-Faure, Camille; Bouillet, Laurence; Hoffmann, Pascale


    On average, 10 % of infertile couples have unexplained infertility. Auto-immune disease (systemic lupus erythematosus, anti-phospholipid syndrome) accounts for a part of these cases. In the last 20 years, aspecific auto-immunity, defined as positivity of auto-antibodies in blood sample without clinical or biological criteria for defined diseases, has been evoked in a subpopulation of infertile women. A systematic review was performed (PUBMED) using the MESH search terms "infertility" and "auto-immunity" or "reproductive technique" or "assisted reproduction" or "in vitro fertilization" and "auto-immunity." We retained clinical and physiopathological studies that were applicable to the clinician in assuming joint management of both infertility associated with serum auto-antibodies in women. Thyroid auto-immunity which affects thyroid function could be a cause of infertility; even in euthyroidia, the presence of anti-thyroperoxydase antibodies and/or thyroglobulin are related to infertility. The presence of anti-phospholipid (APL) and/or anti-nuclear (ANA) antibodies seems to be more frequent in the population of infertile women; serum auto-antibodies are associated with early ovarian failure, itself responsible for fertility disorders. However, there exist few publications on this topic. The methods of dosage, as well as the clinical criteria of unexplained infertility deserve to be standardized to allow a precise response to the question of the role of serum auto-antibodies in these women. The direct pathogenesis of this auto-immunity is unknown, but therapeutic immunomodulators, prescribed on a case-by-case basis, could favor pregnancy even in cases of unexplained primary or secondary infertility.

  15. Prevalence and clinical significance of nonorgan specific antibodies in patients with autoimmune thyroiditis as predictor markers for rheumatic diseases (United States)

    Elnady, Basant M.; Kamal, Naglaa M.; Shaker, Raneyah H.M.; Soliman, Amal F.; Hasan, Waleed A.; Alghamdi, Hamed A.; Algethami, Mohammed M.; Jajah, Mohamed Bilal


    Abstract Autoimmune diseases are considered the 3rd leading cause of morbidity and mortality in the industrialized countries. Autoimmune thyroid diseases (ATDs) are associated with high prevalence of nonorgan-specific autoantibodies, such as antinuclear antibodies (ANA), antidouble-stranded deoxyribonucleic acid (anti-dsDNA), antiextractable-nuclear antigens (anti-ENAs), rheumatoid factor (RF), and anticyclic-citrullinated peptides (anti-CCP) whose clinical significance is unknown. We aimed to assess the prevalence of various nonorgan-specific autoantibodies in patients with ATD, and to investigate the possible association between these autoantibodies and occurrence of rheumatic diseases and, if these autoantibodies could be considered as predictor markers for autoimmune rheumatic diseases in the future. This study had 2 phases: phase 1; in which 61 ATD patients free from rheumatic manifestations were assessed for the presence of these nonorgan-specific autoantibodies against healthy 61 control group, followed by 2nd phase longitudinal clinical follow-up in which cases are monitored systematically to establish occurrence and progression of any rheumatic disease in association to these autoantibodies with its influences and prognosis. Regarding ATD patients, ANA, anti-dsDNA, Anti-ENA, and RF were present in a percentage of (50.8%), (18%), (21.3%), and (34.4%), respectively, with statistically significance difference (P < 0.5) rather than controls. Nearly one third of the studied group (32.8%) developed the rheumatic diseases, over 2 years follow-up. It was obvious that those with positive anti-dsDNA had higher risk (2.45 times) to develop rheumatic diseases than those without. There was a statistically significant positive linear relationship between occurrence of disease in months and (age, anti-dsDNA, anti-CCP, RF, and duration of thyroiditis). Anti-dsDNA and RF are the most significant predictors (P < 0.0001). ATD is more associated with rheumatic

  16. Measuring and evaluating interferon beta-induced antibodies in patients with multiple sclerosis

    DEFF Research Database (Denmark)

    Ross, C; Clemmesen, K M; Sørensen, P S;


    Administration of interferons (IFNs) may induce antibodies that interfere with therapeutic efficacy. We have optimized and validated methods for large-scale economic screening. Sera from patients with relapsing-remitting multiple sclerosis (MS) were investigated for binding antibody (BAb) by prot......Administration of interferons (IFNs) may induce antibodies that interfere with therapeutic efficacy. We have optimized and validated methods for large-scale economic screening. Sera from patients with relapsing-remitting multiple sclerosis (MS) were investigated for binding antibody (BAb...

  17. Comparison of different diagnostic tests for antinuclear antibodies%抗核抗体不同检测方法的对比分析

    Institute of Scientific and Technical Information of China (English)

    武永康; 王兰兰; 唐江涛; 陈捷


    目的 评价以Hep-2细胞为底物的间接免疫荧光法(IFA)和包被多种纯化核抗原的酶联免疫吸附试验(ELISA)检测抗核抗体(ANA)的特点及其在系统性红斑狼疮(SLE)诊断中的应用价值.方法 同时采用IFA法和ELISA法检测血清ANA共409例,其中SLE 226例和健康对照183名,将两种方法检测结果的敏感度、特异度、准确度、阳性预测值、阴性预测值、阳性似然比、阴性似然比、结果符合率、判断一致性κ值及秩相关系数进行比较分析;通过ROC曲线比较两种方法的准确度;结果不一致标本进行可提取性核抗原抗体谱(ENA)检测分析;同时将IFA法检测不同荧光模型样本分别与ELISA结果进行相关分析.结果 SLE患者组IFA法和ELISA法检测ANA的敏感度分别为91.15%和92.04%,特异度分别为96.17%和92.90%,准确度分别为93.40%和92.42%,阳性预测值分别为96.71%和94.12%,阴性预测值分别为89.80%和90.43%,两种方法比较差异均无统计学意义(P>0.05),ROC曲线对两种方法准确度分析差异无统计学意义(P=0.409);两方法检测结果符合率为91.20%,秩相关系数R=0.823,判断一致性κ=0.825;在36例两种方法检测结果不一致标本中,14例IFA(+)ELISA(-),其中1例IFA法滴度达1:1000的高尔基体型,其余为临界值左右的核仁型和斑点型,而在22例IFA(-)ELISA(+)患者中,其中有11例ELISA法吸光度(A)2.67~30.5;将此36例血清标本进行ENA抗体谱检测,14例IFA(+)ELISA(-)患者ENA阳性率为14.29%,而22例IFA(-)ELISA(+)患者ENA阳性率高达68.18%,两者比较差异有统计学意义(P<0.01).IFA法检测的不同荧光模型的阳性标本中,斑点型滴度与ELISA法A值相关性较差R=0.083,均质型相关性最好,R=0.595.结论 IFA作为ANA检测的推荐方法具有直观、可分型等特点,且能提供与自身免疫性疾病相关的核型信息,但需要荧光显微镜及较丰富判断经验的技术人员;而ELISA法操作简单,并可通过测定A值来评价患者体内抗体的浓度,不需再作滴度检测;IFA法和ELISA法检测ANA均有较高的敏感度、特异度、准确性,结果符合率、一致性判断系数κ和秩相关系数均较高,而后者的漏诊率更低,更适合进行ANA筛查;若对ELISA板再增加包被高尔基体、核仁等稀有荧光模型的抗原,则可使ELISA法检测结果更准确、更全面,对申请ANA检测的标本,可先采用ELISA法筛选出阳性标本后再进行IFA法检测,这一工作流程可加快报告速度,节约检验成本,提高工作效率.

  18. 11578例自身抗体检测结果及临床分析%Retrospective Analysis of Anti-nuclear Antibodies Detection in 11578 Patients

    Institute of Scientific and Technical Information of China (English)

    罗雯斌; 姚燕珍; 于倩


    目的 分析浙江省舟山海岛地区抗核抗体和抗核抗体谱阳性分布趋势,及其在就诊人群中的诊断价值.方法 分别用间接免疫荧光法(IIF)和线性免疫印迹法(LIA)检测抗核抗体.资料分布的比较采用x2检验.结果 抗核抗体(ANA)阳性患者总阳性率为16.3% (1886/11578),女性阳性率(20.6%,431/6955)高于男性(9.8%,5/4623),差异有统计学意义(x2=236.7,P<0.05).21 ~40岁组阳性率(1.5%,38/2036)显著高于其他年龄组,差异有统计学意义(x2=115.8,P<0.05).抗核抗体谱阳性患者2722例,阳性率为23.5%.抗核抗体谱中抗SS-A、抗Ro-52、抗着丝点抗体的阳性率最高,分别为31.7%、28.4%和8.3%.女性抗U1-nRNP、抗核糖体P蛋白、抗dsDNA等抗体的阳性率明显高于男性(P<0.05).不同年龄组的抗核抗体谱阳性率分析结果显示抗U1-nRNP、抗Sm、抗核糖体P蛋白等抗体的阳性率差异具有统计学意义(P<0.05).263例不同自身免疫性疾病组中抗核抗体谱阳性率的分析显示RA、SLE、SS、MCTD、PBC组阳性率最高的自身抗体分别是抗Ro-52抗体、抗SS-A抗体、抗SS-B抗体、抗U1-nRNP抗体和抗M-2抗体.结论 舟山海岛地区就诊人群中自身免疫性疾病好发于中年女性,在临床检测中应采用多种自身抗体的联合检测,以提高自身免疫性疾病的诊断率.

  19. 广东中山市2011-2012年HIV抗体检测与自愿咨询检测数据分析%Analysis of HIV antibody screening and VCT data in Zhongshan City,Guangdong Province (2011-2012)

    Institute of Scientific and Technical Information of China (English)

    陈建海; 汪涛; 李雷; 王曼


    目的 了解广东中山市不同人群HIV流行形势,为艾滋病科学防治提供依据.方法 2011-2012年中山市辖区HIV筛查实验室和确诊实验室HIV抗体检测数据,从《中国疾病预防控制信息系统》下载2011-2012年该市自愿咨询检测(VCT)检测数据;采用SPSS进行数据分析.结果 中山市2011-2012年接受HIV筛查累计达77.4万人次,估算每年常住人口中接受HIV筛查的比例约为12.00%.HIV总的筛查阳性率为0.18% (95% CI:0.17%~0.19%),其中男男性行为人群筛查阳性率为10.65%(95% CI:8.56% ~ 12.74%);强制/劳教戒毒人员筛查阳性率为4.30%(95% CI:3.79% ~4.82%),医院一般人群筛查阳性率0.10%(95%CI:0.09%~0.10%).结论 中山市一般人群HIV仍处于低流行阶段,但是男男性行为人群、吸毒人群感染率高.HIV抗体筛查样本量大,为评估不同人群HIV流行形势、疫情估计提供重要参考.%Objective To understand the HIV epidemic characteristics among different groups in Zhongshan city,Guangdong province,to provide information for HIV prevention and control.Methods From 2011 to 2012,the HIV antibody testing data of screening laboratories and confirmed laboratory were collected.The HIV voluntary counseling and testing data were downloaded from < China information system for disease control and prevention >.SPSS was used for data analysis.Results A total of 77.4 million man-time received HIV antibody screening from 2011 to 2012,and the proportion of those receiving HIV screening among the permanent inhabitant was estimated to be 12.00% each year.The overall HIV antibody screening positive rate was 0.18% (95% CI:0.17%-0.19%).The HIV antibody screening positive rate of men who had sex with men,drug users of compulsory/drug treatment center,and common people visiting hospitals were 10.65% (95% CI:8.56%-12.74%),4.30% (95% CI:3.79%-4.82%),and 0.10% (95% CI:0.09%-0.10

  20. Screening Anti-LPS VHH from Llama Single Domain Antibody Li-brary and Preparation of Anti-LPS Nanobody Pentamer%抗脂多糖纳米抗体的筛选及其五聚体的制备

    Institute of Scientific and Technical Information of China (English)

    武文; 李胜华; 张伟京; 王炳奇


    目的::筛选抗脂多糖(LPS)纳米单域抗体,并制备抗LPS纳米抗体五聚体。方法:以LPS为抗原,从驼源天然单域重链抗体库中筛选抗LPS纳米抗体,利用分子克隆技术将抗LPS单域抗体基因组装入志贺杆菌样毒素B亚基蛋白结构域(VTB)的五聚体特异性表达载体中进行可溶性表达,并用ELISA法鉴定所获抗体的抗原结合活性和特异性。结果:获得抗LPS纳米单域抗体及LPS纳米抗体五聚体;经鉴定,LPS纳米抗体五聚体的抗原结合活性优于抗LPS单域抗体。结论:利用驼源天然单域重链抗体库制备了抗LPS纳米单域抗体及抗LPS纳米抗体五聚体,为脓毒血症的分子诊断、预后判断及寻找生物治疗新靶点奠定了基础。%Objective: To obtain anti-LPS VHH and prepare anti-LPS nanobody pentamer. Methods: Using phage display screening technology, we selected anti-LPS single domain antibody(sdAb, or nanobody) from a phage display library of llama single domain antibody, and then cloned the anti-LPS single-domain antibody gene into specific expression vector pVTB1 with molecular cloning technology, and expressed anti-LPS pentamer effi-ciently in E.coli. The affinity of different type of anti-LPS nanobodies were detected by ELISA. Results: We ob-tained anti-LPS single domain antibody and anti-LPS antibody pentamer, and proved anti-LPS nanobody pentamer is extremely enhance the specificity and affinity strength of anti-LPS antibody. Conclusion: The selected anti-LPS antibody pentamer is a leading candidate for therapies against LPS-mediated sepsis.

  1. Screening for toxoplasmosis in pregnancy. (United States)

    Broadbent, E J; Ross, R; Hurley, R


    The prevalence of antibody against Toxoplasma gondi in a population of 715 pregnant women has been evaluated by two methods: indirect haemagglutination antibody (IHA) and indirect fluorescent antibody (IFA) test and all positive sera were checked by the dye test. Five hundred of the study population were questioned on diet and on animal contact to elucidate a possible relation to the prevalence of antibody. Results are expressed in international units (IU) of antibody against T gondi. Of the 715 sera, 171 were positive by IHA and 173 by IFA. One hundred and sixty-seven sera were positive by both tests, ninety-eight (58%) correlating exactly, as to the concentration of antibody. The ten sera which were not positive by both tests all had detectable antibody at the minimum concentration only (12 IU). The dye test confirmed all sera positive by both tests with the exception of three. It also confirmed one of four sera positive by IHA antibody alone and two of six positive by IFA alone. All sera that proved dye test-negative had low antibody concentrations (12 IU) by IHA or IFA. The IHA test, which is commercially available in kit form, would be suitable for use as a screening test during pregnancy. The estimated annual rate of antibody acquisition over the age range 16-40 years is 1.2% per annum with the highest rate in the 36-40 age group (2.5% per annum) and the lowest in the 26-30 age group (0.4% per annum). The clinical history was not significantly different between those with and those without antibody against T gondi but significantly more women in the 36-40 age group had a history of animal contact than those in the 26-30 age group. No conclusive evidence of recent or current infection was found.

  2. Depression Screening (United States)

    ... Centers Diseases + Condition Centers Mental Health Medical Library Depression Screening (PHQ-9) - Instructions The following questions are ... this tool, there is also text-only version . Depression Screening - Manual Instructions The following questions are a ...

  3. Cancer Screening

    Directory of Open Access Journals (Sweden)

    Krishna Prasad


    Full Text Available Cancer screening is a means to detect cancer early with the goal of decreasing morbidity and mortality. At present, there is a reasonable consensus regarding screening for breast, cervical and colorectal cances and the role of screening is under trial in case of cancers of the lung,  ovaries and prostate. On the other hand, good screening tests are not available for some of the commonest cancers in India like the oral, pharyngeal, esophageal and stomach cancers.

  4. Autoimmune encephalitis: Clinical diagnosis versus antibody confirmation

    Directory of Open Access Journals (Sweden)

    Asha Caroline Cyril


    Full Text Available Context: Autoimmune encephalitis is a heterogeneous disorder which is being diagnosed with increasing frequency. The diagnosis of these disorders is based on the detection of autoantibodies and characteristic clinical profiles. Aims: We aimed to study the antibody profile in encephalitis patients with suspected autoimmune etiology presenting to a tertiary care center. Settings and Design: The subjects were selected by screening all patients with clinical profile suggesting autoimmune encephalitis admitted in the neuromedical intensive care unit (ICU of a tertiary care center in South India. Materials and Methods: Patients who fulfilled modified Zuliani et al.′s, criteria for autoimmune encephalitis were identified during the period December 2009-June 2013. Blood samples from these subjects were screened for six neuronal antibodies. Statistical analysis used: Chi-square test was applied to compare the antibody positive and negative patients. Results: Out of 1,227 patients screened, 39 subjects (14 males: 25 females were identified with a mean age of 15.95 years and 19 cases were assessed in the acute and 20 in the convalescent phase of the illness. Seizure (87.8 % was the most common presenting symptom; status epilepticus occurred in 23 (60.5% patients during the course of the illness. Fourteen (35.9% patients were N-methyl-D-aspartate receptor (NMDAR antibody-positive and all were negative for the other antibodies tested. Conclusions: One-third of patients presenting with acute noninfective encephalitis would be positive for NMDAR antibodies with the remaining two-thirds with clinically suspected autoimmune encephalitis being antibody-negative. There are few markers in the clinical and investigative profiles to distinguish antibody-positive and -negative patients.

  5. Colon cancer screening (United States)

    Screening for colon cancer; Colonoscopy - screening; Sigmoidoscopy - screening; Virtual colonoscopy - screening; Fecal immunochemical test; Stool DNA test; sDNA test; Colorectal cancer - screening; Rectal ...


    Serum samples from 251 wild carnivores from different regions of Spain were tested for antibodies to Neospora caninum by the commercial competitive screening enzyme linked immunosorbent assay (c-ELISA) and confirmed by Neospora agglutination test (NAT) and/or by indirect fluorescent antibody test (I...

  7. Monoclonal antibody technologies and rapid detection assays (United States)

    Novel methodologies and screening strategies will be outlined on the use of hybridoma technology for the selection of antigen specific monoclonal antibodies. The development of immunoassays used for diagnostic detection of prions and bacterial toxins will be discussed and examples provided demonstr...

  8. Selection of antibodies from synthetic antibody libraries. (United States)

    Harel Inbar, Noa; Benhar, Itai


    More than 2 dozen years had passed since the field of antibody engineering was established, with the first reports of bacterial [1-3] and mammalian cells [4] expression of recombinant antibody fragments, and in that time a lot of effort was dedicated to the development of efficient technological means, intended to assist in the creation of therapeutic monoclonal antibodies (mAbs). Research focus was given to two intertwined technological aspects: the selection platform and the recombinant antibody repertoires. In accordance with these areas of interest, it is the goal of this chapter to describe the various selection tools and antibody libraries existing, with emphasis on the later, and their applications. This chapter gives a far from exhaustive, subjective "historic account" of the field, describing the selection platforms, the different formats of antibody repertoires and the applications of both for selecting recombinant antibodies. Several excellent books provide detailed protocols for constructing antibody libraries and selecting antibodies from those libraries [5-13]. Such books may guide a newcomer to the field in the fine details of antibody engineering. We would like to offer advice to the novice: although seemingly simple, effective library construction and antibody isolation provide best benefits in the hands of professionals. It is an art as much as it is science.

  9. Prevalence and chemotherapy-induced reactivation of occult hepatitis B virus among hepatitis B surface antigen negative patients with diffuse large B-cell lymphoma: Significance of hepatitis B core antibodies screening

    Directory of Open Access Journals (Sweden)

    Tamer A. Elbedewy


    Conclusion: The study concluded that anti-HBc screening is mandatory before chemotherapy. HBsAg-negative/anti-HBc-positive patients should be closely observed for signs of HBV reactivation through the regular monitoring of ALT. Prophylaxis lamivudine is recommended for anti-HBc positive patients before chemotherapy.

  10. Are the current attempts at standardization of antiphospholipid antibodies still useful? Emerging technologies signal a shift in direction. (United States)

    Andreoli, Laura; Rizzini, Silvia; Allegri, Flavio; Meroni, Pierluigi; Tincani, Angela


    The pathogenic role of antiphospholipid antibodies (aPL) has been widely established over past years in several experimental models and clinical studies. Accordingly, the detection of aPL by immunoassays (anticardiolipin antibodies; anti-beta2 glycoprotein I antibodies) has become a routine practice in the clinical workup of patients with systemic autoimmune diseases. aPL are mostly assayed using commercial ELISA kits, whose performance has not been found to be sufficiently concordant among the different manufacturers. In the past years, collaborative groups have spent considerable effort to reach some form of standardization but this process is still ongoing. Such lack of standardization has recently become even more crucial, as manufacturers have had to face an increasing demand for fully automated tests for aPL, like those test systems that have been developed for other autoantibodies (e.g., antinuclear antibodies, anti-ENA antibodies). We therefore report our recent experience with two newly developed automated methods for anticardiolipin antibodies testing. In particular, we discuss the results obtained using routine samples, as we believe that these better reflect the "real-life" situation in which those automated methods will operate. We also mention other emerging technologies in the field of aPL detection.

  11. 人源Fab抗体库的构建和抗hPRLR抗体的筛选鉴定%Screening and identification of human Fab antibody against hPRLR from large phage-display library originated from breast cancer patients

    Institute of Scientific and Technical Information of China (English)

    屈芫; 魏钦俊; 姚俊; 鲁雅洁; 王天明; 曹新; 冯振卿


    cWe aim to get specific Fab antibody against human prolactin receptor (hPRLR) from the human Fab antibody library constructed by phage display technology. Human lymphocytes were collected from peripheral blood of 24 breast cancer patients. And then the total RNA was extracted and reversely transcribed to cDNA. Genes of light and heavy chains of human antibody were amplified by RT-PCR to construct anti-hPRLR immunized human antibody library. After three rounds of panning and one round of crossed-panning with against his-hPRLR fusion protein, BSA-polypeptide (epitopes of hPRLR) and GST-hPRLR fusion protein, positive clones were chosen by Phage-ELISA and DNA sequencing. Then the positive clones were transformed into E. coli Top 10' and induced to express antibody protein. The results indicated that the human Fab phage-display library consisting of l.0×l09 clones were successfully constructed, and six clones were selected after four rounds. One of them named FabG2 expressed protein correctly. ELISA and Western blot analysis showed that FabG2 could bind hPRLR specifically. We concluded that the hPRLR specific Fab antibody selected from large phage-display library could be used as candidates for therapy agent of breast cancer which over-expresses hPRLR.%目的 构建人源Fab噬菌体抗体库,筛选抗hPRLR抗体片段并进行初步鉴定.方法 从乳腺癌患者外周血提取总RNA,通过RT-PCR扩增人抗体轻链和重链基因,构建抗hPRLR人源Fab抗体库.分别以His-hPRLR融合蛋白、BSA-hPRLR表位多肽融合蛋白和GST-hPRLR融合蛋白作为抗原包板,经过3轮循环的吸附一洗脱一扩增的筛选及1轮交叉筛选,挑单克隆用Phage-ELISA、DNA测序筛选阳性克隆,将筛选到的阳性克隆Fabc2转化至Top10’受体菌,诱导表达可溶性Fab抗体,通过Western blot和ELISA进行特异性的鉴定.结果 构建的人源Fab库容为1.0×109,4轮的筛选,获得6株能与hPRLR结合的人源抗体克隆.选取的Fabc2能够进行

  12. Screening CO

    NARCIS (Netherlands)

    Ramírez, A.; Hagedoorn, S.; Kramers, L.; Wildenborg, T.; Hendriks, C.


    This paper describes the development and application of a methodology to screen and rank Dutch reservoirs suitable for long-term large scale CO2 storage. The screening focuses on off- and on-shore individual aquifers, gas and oil fields. In total 176 storage reservoirs have been taken int

  13. Neonatal molecular pathologies induced by maternal anti-Ro and anti-La antibodies

    Directory of Open Access Journals (Sweden)

    Herrera-Esparza R


    Full Text Available Maternal antinuclear antibodies with anti-Ro or anti-La specificity might be pathogenic to the fetus and could induce molecular neonatal pathologies, such as neonatal lupus (NL with or without congenital heart block (CHB. The cutaneous manifestations of neonatal lupus appear at birth or a few weeks later, and skin lesions may persist for weeks. While CHB is characterized by intrauterine bradycardia or low heart rates at birth and may persist for months, depending on the degree of blockage. Clinical and experimental data demonstrated that anti-Ro and anti-La autoantibodies functionally inhibit L-type calcium channels and induce abnormalities in electrical conduction of the cardiac myocytes. It has been 38 years since the first clinical description of CHB. Presently, the pathophysiology of CHB has been clarified through clinical and basic research studies.

  14. The significance of antiphospholipid antibodies in pregnant women with chronic hypertension

    NARCIS (Netherlands)

    Zeeman, GG; Alexander, JM; McIntire, DD; Leveno, KJ


    The objective of this study was to perform antiphospholipid antibody screening in women with chronic hypertension to assess whether the presence of such antibodies is associated with adverse pregnancy outcome. Serum for anticardiolipin antibodies and lupus anticoagulant was obtained in pregnant wome


    NARCIS (Netherlands)



    Antibodies against ribosomal phosphoproteins (anti-P antibodies) are found in about 10% of patients with systemic lupus erythematosus (SLE). Using an ELISA with a synthetic peptide for screening and an immunoblotting technique as a confirmation test for detection of these antibodies, we found 16 pos

  16. Antibody-based resistance to plant pathogens. (United States)

    Schillberg, S; Zimmermann, S; Zhang, M Y; Fischer, R


    Plant diseases are a major threat to the world food supply, as up to 15% of production is lost to pathogens. In the past, disease control and the generation of resistant plant lines protected against viral, bacterial or fungal pathogens, was achieved using conventional breeding based on crossings, mutant screenings and backcrossing. Many approaches in this field have failed or the resistance obtained has been rapidly broken by the pathogens. Recent advances in molecular biotechnology have made it possible to obtain and to modify genes that are useful for generating disease resistant crops. Several strategies, including expression of pathogen-derived sequences or anti-pathogenic agents, have been developed to engineer improved pathogen resistance in transgenic plants. Antibody-based resistance is a novel strategy for generating transgenic plants resistant to pathogens. Decades ago it was shown that polyclonal and monoclonal antibodies can neutralize viruses, bacteria and selected fungi. This approach has been improved recently by the development of recombinant antibodies (rAbs). Crop resistance can be engineered by the expression of pathogen-specific antibodies, antibody fragments or antibody fusion proteins. The advantages of this approach are that rAbs can be engineered against almost any target molecule, and it has been demonstrated that expression of functional pathogen-specific rAbs in plants confers effective pathogen protection. The efficacy of antibody-based resistance was first shown for plant viruses and its application to other plant pathogens is becoming more established. However, successful use of antibodies to generate plant pathogen resistance relies on appropriate target selection, careful antibody design, efficient antibody expression, stability and targeting to appropriate cellular compartments.

  17. Blame for climatic catastrophe. An unholy alliance of reactor engineers and antinuclear campaigners; Schuld an der Klimakatastrophe. Eine unheilige Allianz von Reaktorbauern und Kernkraftgegnern

    Energy Technology Data Exchange (ETDEWEB)

    Frenk, H.


    May be that dedicated climate protectors harm the climate than protect it? LWRs should'nt play a role in future because of their remaining risk but HTRs without this risk should be promoted and can so reduce CO2 emission. But envy of concurrence of the LWR lobby and dogmatic stubborness of the antinuclear campaigners take steps against this nonpolluting perspective. [German] Kann es sein, dass engagierte Klimaschuetzer dem Klima eher schaden als nuetzen? Helmuth Frenk bejaht diese Frage, denn nach seiner Erkenntnis wirken die Aktionen der Kernkraftgegner mit ihrer Ablehnung jeglicher Atomkraftnutzung kontraproduktiv. Waehrend Leichtwasserreaktoren (LWR) wegen des erhoehten atomaren Restrisikos kaum fuer einen verstaerkten Einsatz zur Minderung des Kohlendioxidausstosses in Frage kommen, sollten die risikofreien Hochtemperaturreaktoren (HTR) gefoerdert werden. Denn sie wuerden mit der vielfachen Effizienz der LWR, erst recht aller alternativen Stromerzeuger, die CO{sub 2}-Emission reduzieren. Konkurrenzneid der LWR-Lobby und dogmatische Verbohrtheit der Kernkraftgegner stehen dieser umweltfreundlichen Perspektive entgegen. (orig.)

  18. Structure Based Antibody-Like Peptidomimetics

    Directory of Open Access Journals (Sweden)

    Mark I. Greene


    Full Text Available Biologics such as monoclonal antibodies (mAb and soluble receptors represent new classes of therapeutic agents for treatment of several diseases. High affinity and high specificity biologics can be utilized for variety of clinical purposes. Monoclonal antibodies have been used as diagnostic agents when coupled with radionuclide, immune modulatory agents or in the treatment of cancers. Among other limitations of using large molecules for therapy the actual cost of biologics has become an issue. There is an effort among chemists and biologists to reduce the size of biologics which includes monoclonal antibodies and receptors without a reduction of biological efficacy. Single chain antibody, camel antibodies, Fv fragments are examples of this type of deconstructive process. Small high-affinity peptides have been identified using phage screening. Our laboratory used a structure-based approach to develop small-size peptidomimetics from the three-dimensional structure of proteins with immunoglobulin folds as exemplified by CD4 and antibodies. Peptides derived either from the receptor or their cognate ligand mimics the functions of the parental macromolecule. These constrained peptides not only provide a platform for developing small molecule drugs, but also provide insight into the atomic features of protein-protein interactions. A general overview of the reduction of monoclonal antibodies to small exocyclic peptide and its prospects as a useful diagnostic and as a drug in the treatment of cancer are discussed.

  19. 抗核抗体及抗核抗体谱的实验检测方法及结果分析%The Analysis of Antinuclear Antibodies and Antinuclear Antibody Spectrum Assay Method and Result of Autoimmune Dis-eases

    Institute of Scientific and Technical Information of China (English)

    朱凤丹; 尹秀杉


    Objective :To explore the value and results that using indirect immunofluorescence and immunoblottingto detect ANA and ANAs .Methods :Analysis the 2 317 cases were grouped by age ,sex and the results of ANA and AN‐As .Results:In this 2 317 cases ,the positive of ANA and/or ANAs were 914 cases (39 .4% ) ,women accounted for 77.7% ,males accounted for 22 .3% ,36 years and over age group ,women accounted for 50 .1% ,accounting for 16 .1%of men;ANA positive 740 cases (31 .9% ) ,ANAs tested positive for 723 cases (31 .2% ) ,ANA and ANAs were posi‐tive in 549 cases ,ANA (+ ) and ANAs (+ ) in line with the rate of 74 .2% ,ANA (-) and ANAs (-) compliance rate of 89 .0% ,the overall compliance rate of 84 .2% .Conclusion:Combined detection of ANA and ANAs ,not only can reduce the false‐negative and false‐positive rate ,but also provide the basis for more effective laboratory diagnosis and differential diagnosis of autoimmune disease .%目的:探讨分析实验室采用间接免疫荧光法和免疫印迹法检测ANA和ANAs的结果及价值。方法:对2317例自身抗体受检者的年龄、性别及ANA和ANAs检测结果进行分析。结果:在2317例受检者中,ANA和/或ANAs阳性914例(39.4%),女性占77.7%,男性占22.3%,36岁及以上年龄段,女性占50.1%,男性的占16.1%;A N A阳性740例(31.9%),ANAs阳性723例(31.2%),ANA 和 ANAs 均阳性549例,ANA (+)和 ANAs (+)符合率74.2%,ANA(-)和ANAs(-)符合率89.0%,总体符合率84.2%。结论:联合检测ANA和ANAs ,不仅可以减低假阴性率和假阳性率,而且能为诊断和鉴别诊断自身免疫病提供更有力的实验室依据。

  20. Quadruple screen test (United States)

    Quad screen; Multiple marker screening; AFP plus; Triple screen test; AFP maternal; MSAFP; 4-marker screen; Down syndrome - quadruple; Trisomy 21 - quadruple; Turner syndrome - quadruple; Spina bifida - ...

  1. Hypertension screening (United States)

    Foulke, J. M.


    An attempt was made to measure the response to an announcement of hypertension screening at the Goddard Space Center, to compare the results to those of previous statistics. Education and patient awareness of the problem were stressed.

  2. Airport Screening (United States)

    ... cannot become radioactive from this procedure. However, some photographic film may need to be hand-screened because ... level. An American National Standards Institute/Health Physics Society industry standard states that the maxi- mum allowable ...

  3. Toxicology screen (United States)

    Toxicology screening is most often done using a blood or urine sample. However, it may be done soon after the person swallowed the medication, using stomach contents taken through gastric lavage (stomach pumping) or after vomiting.

  4. Towards Universal Screening for Toxoplasmosis: Rapid, Cost-Effective, and Simultaneous Detection of Anti-Toxoplasma IgG, IgM, and IgA Antibodies by Use of Very Small Serum Volumes. (United States)

    Augustine, Swinburne A J


    Rapid, cost-effective, and early determination of the serological status of potentially infected individuals, particularly pregnant women, can be critical in preventing life-threatening infections and subsequent fetal congenital abnormalities. An article in this issue of the Journal of Clinical Microbiology (X. Li, C. Pomares, G. Gonfrier, B. Koh, S. Zhu, M. Gong, J. G. Montoya, and H. Dai, J Clin Microbiol 54:1726-1733, 2016, describes an innovative multiplexed immunoassay that offers a path toward universal screening.

  5. Construction and screening of phage display single chain antibody library against histidine-rich protein Ⅱ of Plasmodium falciparum%抗恶性疟原虫富含组氨酸蛋白Ⅱ单链抗体库的构建及筛选

    Institute of Scientific and Technical Information of China (English)

    侯云霞; 董文其; 徐伟文; 王萍; 陈白虹; 李明


    Objective To construct phage display single-chain antibody fragments (scFvs) library against histidine-rich protein Ⅱ (HRP-Ⅱ) of Plasmodium falciparum and select specific scFvs of anti- HRP-Ⅱ for the purpose of malaria diagnosis. Method The genes of variable fragments of heavy chain (VH) and light chain (VL) were gained from the spleen cells of BALB/c mice immunized with HRP-Ⅱ protein. The VH and VL genes were then assembled by the method of splicing overlapping extension and cloned into phagemid vector pCANTAB 5E. The scFv phage antibodies were expressed at the surface of the phage after the rescue by helper phage M13K07. HRP- Ⅱ protein was used as antigenic reagent for panning and screening. Results The total RNA from the spleen cells was isolated, and cDNA obtained and VH and VL gene regions amplified using PCR. The VH and VL gene regions were combined with a flexible linker ligated into the pCANTAB 5E phagemid vector, and transformed into TG1 Escherichia coli. The repertoire of the phage antibody was about 106. After panning and screening, 8 positive clones expressed scFv antibodies which were specific for HPR-Ⅱ as demonstrated by ELISA. Conclusion Phage display technology can be used as a powerful tool in making scFv antibodies which have the potential to be used as reagents in the diagnosis and therapy of malaria.%目的构建抗恶性疟原虫富含组氨酸蛋白Ⅱ(Histidine- rich protein II, HRP-II)单链抗体库,并筛选出阳性克隆。方法用噬菌体抗体库技术构建抗恶性疟原虫HRP-Ⅱ单链抗体库,并以HRP-Ⅱ为靶抗原对该库进行了三轮"亲和吸附-援救-感染扩增"的富集,挑取单菌落筛选并鉴定阳性克隆。结果获得目的基因并成功构建抗恶性疟原虫HRP-Ⅱ的单链抗体库,库容为106,并从中筛选出8株阳性克隆。结论噬菌体抗体库技术具有高效的筛选性能,抗 HRP-Ⅱ单链抗体的制备为其在恶性疟的免疫快速诊断方法中的应用奠定了基础。

  6. The antibody mining toolbox: an open source tool for the rapid analysis of antibody repertoires. (United States)

    D'Angelo, Sara; Glanville, Jacob; Ferrara, Fortunato; Naranjo, Leslie; Gleasner, Cheryl D; Shen, Xiaohong; Bradbury, Andrew R M; Kiss, Csaba


    In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput screening methods such as enzyme-linked immunosorbant assay. The high cost and the need for bioinformatics experts and powerful computer clusters, however, have limited the general use of deep sequencing in antibody selections. Here, we describe the AbMining ToolBox, an open source software package for the straightforward analysis of antibody libraries sequenced by the three main next generation sequencing platforms (454, Ion Torrent, MiSeq). The ToolBox is able to identify heavy chain CDR3s as effectively as more computationally intense software, and can be easily adapted to analyze other portions of antibody variable genes, as well as the selection outputs of libraries based on different scaffolds. The software runs on all common operating systems (Microsoft Windows, Mac OS X, Linux), on standard personal computers, and sequence analysis of 1-2 million reads can be accomplished in 10-15 min, a fraction of the time of competing software. Use of the ToolBox will allow the average researcher to incorporate deep sequence analysis into routine selections from antibody display libraries.

  7. Acetylcholine receptor antibody (United States)

    ... page: // Acetylcholine receptor antibody To use the sharing features on this page, please enable JavaScript. Acetylcholine receptor antibody is a protein found in the blood ...

  8. Lyme disease antibody (United States)

    ... JavaScript. The Lyme disease blood test looks for antibodies in the blood to the bacteria that causes ... needed. A laboratory specialist looks for Lyme disease antibodies in the blood sample using the ELISA test . ...

  9. New monoclonal antibodies directed against human renin. Powerful tools for the investigation of the renin system.


    Galen, F X; Devaux, C.; Atlas, S; Guyenne, T; Menard, J; Corvol, P; Simon, D.; Cazaubon, C; Richer, P; Badouaille, G


    Monoclonal antibodies directed against human renin were obtained by the fusing of myeloma cells with spleen cells from Balb/c or high-responder Biozzi mice injected with pure tumoral or highly purified renal renin. These procedures resulted in the production of seven stable monoclonal antibodies to human renin. Antibodies in the hybridoma culture medium were screened by binding to pure iodinated renin or insolubilized renin in a solid phase assay. The concentration of purified antibodies that...

  10. The antibody mining toolbox


    D'Angelo, Sara; Glanville, Jacob; Ferrara, Fortunato; Naranjo, Leslie; Gleasner, Cheryl D.; Shen, Xiaohong; Bradbury, Andrew RM; Kiss, Csaba


    In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput ...

  11. HCC screening; HCC-Screening

    Energy Technology Data Exchange (ETDEWEB)

    Albrecht, T. [Charite-Unversitaetsmedizin,Freie Universitaet und Humboldt-Universitaet zu Berlin, Klinik und Hochschulambulanz fuer Radiologie und Nuklearmedizin,Campus Benjamin Franklin, Berlin (Germany)


    Hepatocellular carcinoma (HCC) is one of the most frequently diagnosed tumour diseases throughout the world. In the vast majority of cases those affected are high-risk patients with chronic viral hepatitis and/or liver cirrhosis, which means there is a clearly identifiable target group for HCC screening. With resection, transplantation, and interventional procedures for local ablation, following early diagnosis curative treatment options are available with which 5-year survival rates of over 60% can be reached. Such early diagnosis is a reality only in a minority of patients, however, and in the majority of cases the disease is already in an advanced stage at diagnosis. One of the objects of HCC screening is diagnosis in an early stage when curative treatment is still possible. Precisely this is achieved by screening, so that the proportion of patients treated with curative intent is decisively higher. There is not yet any clear evidence as to whether this leads to a lowering of the mortality of HCC. As lower mortality is the decisive indicator of success for a screening programme the benefit of HCC screening has so far been neither documented nor refuted. Nonetheless, in large regions of the world it is the practice for high-risk patients to undergo HCC screening in the form of twice-yearly ultrasound examination and determination of AFP. (orig.) [German] Das hepatozellulaere Karzinom (HCC) ist eine der weltweit haeufigsten Tumorerkrankungen. Es tritt in der grossen Mehrzahl der Faelle bei Hochrisikopatienten mit chronischer Virushepatitis bzw. Leberzirrhose auf, woraus sich eine klar identifizierbare Zielgruppe fuer das HCC-Screening ergibt. Mit der Resektion, der Transplantation und interventionellen lokal ablativen Verfahren stehen bei rechtzeitiger Diagnosestellung kurative Therapieoptionen zur Verfuegung, die 5-Jahres-Ueberlebensraten von >60% erreichen. Diese rechtzeitige Diagnosestellung erfolgt jedoch nur bei einer Minderzahl der Patienten, waehrend die

  12. Analysis of positive anti-ENA antibodies comparing with their antinuclear antibodies in serum%抗可提取性核抗原抗体与抗核抗体的对照分析

    Institute of Scientific and Technical Information of China (English)



    目的 探讨抗可提取性核抗原(ENA)多肽抗体不同项目阳性时与抗核抗体(ANA)之间的相关性.方法 对比分析 323例抗ENA抗体(抗U1-nRNP、Sm、SS-A、SS-B、Scl-70和Jo-1抗体)阳性病例的多肽谱类型与其304例ANA阳性核型.结果 抗ENA抗体类型不同,ANA荧光核型有很大差别,没有绝对的规律可言.结论 不能简单地依据荧光核型来推断抗ENA抗体的具体类型.

  13. 抗核抗体荧光核型与特异性抗体相关性分析%Correlation between antinuclear antibody fluorescence patterns and specific antibodies

    Institute of Scientific and Technical Information of China (English)

    吴丽娜; 郭萍; 丁宁


    目的 探讨抗核抗体荧光核型与特异性抗体之间的相关性,以期为临床提供更优、更经济的抗核抗体检测组合方式.方法 应用欧蒙间接免疫荧光法(IIFA)检测抗核抗体(ANA)判断荧光核型,欧蒙印迹法(LIA)抗核抗体谱(ANAs)15项检测特异性抗体.对3 783例患者IIFA-ANA、LIA-ANAs结果进行统计分析,根据检测结果分为IIFA-ANA+/LIA-ANAs-组1 548例、IIFA-ANA+/LIA-ANAs+组2 220例、IIFA-ANA-/LIA-ANAs+组15例.结果 3768例IIFA-ANA+患者,LIA-ANAs的阳性率为58.92%;1 704例IIFA-ANA+/LIA-ANAs+单一荧光核型患者,IIFA-ANA与LIA-ANAs的符合率分别为核颗粒型82.61%、核均质82.80%、核仁型82.86%、着丝点型100%、胞浆型69.29%.IIFA-ANA-/LIA-ANAs+组降低IIFA-ANA稀释倍数后,出现与特异性抗体相对应的荧光核型.结论 不能简单依靠IIFA-ANA的荧光核型推断特异性抗体;临床医生可以根据患者病情、IIFA-ANA荧光核型决定患者是否需要检测LIA-ANAs.

  14. Karyotype Distribution of Antinuclear Antibodies and Its Correlation with Specific Antibody%抗核抗体核型分布及核型与特异性抗体相关性分析

    Institute of Scientific and Technical Information of China (English)

    李正; 陈杰; 袁文娟; 周珣


    目的 通过对ANA核型分布及核型与特异性抗体相关性分析,为临床对ANA和ANA谱检测结果判读提供参考.方法 用间接免疫荧光法检测ANA,免疫印迹法检测ANA谱,对449例ANA或ANA谱阳性的结果进行核型和特异性抗体分析.结果 ANA核型分布以核颗粒型为主,占78.2%;核型与特异性抗体有一定的相关性,但同一种自身抗体可出现不同的荧光核型.结论 ANA和ANA谱同时检测能提高AID的检出率,有助于准确的判断特异性抗体,对AID的诊断、病情监测、预后判断有十分重要的作用.

  15. Construction of a human na(i)ve Fab library and screening of phage antibody against arginine vasopressin%人源Fab噬菌体抗体库的构建与抗精氨酸加压素抗体的筛选

    Institute of Scientific and Technical Information of China (English)

    董越华; 胡占东; 公倩; 李常颖; 畅继武; 朱铁虹


    Objective: To construct a naive human Fab phage display library, screen and identify arginine vasopressin Fab antibody from the library. Methods: Total RNA was extracted from peripheral blood lymphocytes of 18 healthy donors, and the light chain and heavy chain Fd genes were amplified by RT-PCR. Then the amplification products were sequentially cloned into phagemid vector pComb3XSS to construct a human Fab phage antibody library. The insertion of the light chain or heavy chain Fd genes were identified by cutting with endonucleases and PCR amplification. Arginine vasopressin was used as target antigen to pan the original Fab antibody library. After five rounds of panning were carried out,fifty randomly selected clones were assayed by phage-ELISA analysis. The positive clones were analyzed by DNA sequencing. Results: A large human Fab phage antibody library consisting of 2.4 × 108 members was successfully constructed. After having been panned by AVP,we obtained six positive clones which had specificity and binding reactivity towards AVP. The C4 clone was analyzed and showed that its heavy chain belonged to IgG subvariety and its light chain to X family. Conclusion : We successfully constructed a large human Fab phage antibody library and isolated the specific human anti-AVP Fab antibodies,which provided a solid foundation for the establishment of rapid detection method of arginine vasopressin in future research.%目的:构建人源天然Fab噬菌体抗体库,筛选抗精氨酸加压素特异性抗体并进行初步鉴定.方法:从18位健康成人的外周血淋巴细胞,提取总RNA.利用RT-PCR扩增人Fab抗体基因片段,将其克隆至噬菌粒载体pComb3XSS内,构建人源天然Fab噬菌体抗体库.以固相化的精氨酸加压素为靶抗原对抗体库进行五轮筛选后,随机挑取50个单克隆进行phage-ELISA检测,阳性克隆行DNA测序分析.结果:成功构建库容为2.4×108的噬菌体抗体库,从中筛选到6株阳性克隆能够与精

  16. Screening of neutralizing antibody titers against enterovirus 71 in human intravenous immunoglobulin ( pH 4 )%静注人免疫球蛋白(pH4)中肠道病毒71型中和抗体效价的筛查

    Institute of Scientific and Technical Information of China (English)

    王敏力; 王威; 孙思才; 丁勇; 侯继锋


    Objective To screen the neutralizing antibody titers against enterovirus 71 (EV71) in commercial human intravenous immunoglobulin (IVIG)(pH 4) and experimental intravenous EV71 immunoglobulin,and provide a basis for further passive immunotherapy of hand-foot-mouth disease (HFMD). Methods The neutralizing antibody titers against EV71 in domestic 1VIG (pH 4) and in experimental intravenous EV71 immunoglobulin prepared with screened raw plasma were determined by microcytopathic effect (micro-CPE) method with two virus strains in two laboratories. Results The neutralizing antibody titers of 55 batches of domestic IVIG manufactured by 17 manufacturers determined with strain-01 in laboratory A were 104 ~ 256,of which 23. 64% (13/55) were not less than 239,12. 73% (7/55) were not less than 256,while the GMTs of neutralizing antibodies of 3 batches of experimental intravenous EV71 immunoglobulin were 1 203. However,the GMTs of neutralizing antibody against EV71 in 3 batches of experimental intravenous EV71 immunoglobulin (1 402) determined with strain-01 in NIFDC laboratory were significantly higher than those of 12 batches of IVIG manufactured by 8 manufacturers (P 0. 05). Conclusion All the neutralizing antibody against EV71 in domestic IVIG manufactured by various manufacturers were positive,of which the titers ranged from 104 to 630. However,the neutralizing antibody titers against EV71 in intravenous EV71 immunoglobulin prepared with the screened plasma were significantly higher than those in IVIG.%目的 对目前市售静注人免疫球蛋白(Human intravenous immunoglobulin,简称IVIG)(pH 4)以及静注肠道病毒71型(Enterovirus71,EV71)免疫球蛋白试验品中的EV71中和抗体效价进行筛查,为手足口病(Hand-foot-mouth disease,HFMD)的被动免疫治疗提供参考.方法 采用微量细胞病变法,在2个实验室,采用2个毒株检测国内IVIG产品以及经原料血浆筛查后制备的静注EV71免疫球蛋白试验品的EV71

  17. Update: routine screening for antibodies to human immunodeficiency virus, civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components, January 2008-June 2013. (United States)


    This report contains an update through June 2013 on the results of screening for HIV infection among civilian applicants for military service and among members of the active and reserve components of the Armed Forces. Among civilian applicants, annual rates of prevalence of HIV infection showed a continuing downward trend. Rates among black, non-Hispanic applicants were higher than other racial/ethnic groups but have declined sharply since 2008. Among service members, annual rates have varied by service and component, with higher rates in the Army and Navy and lower rates in the Marine Corps and Air Force. Members of the Army and Air Force Reserves have had consistently higher rates than members of their respective active components. For both civilian applicants and service members, rates among men are notably higher than among women. The possible roles of unprotected sex and pre-deployment behaviors and the associated challenges to prevention of HIV infection are discussed.

  18. Generation and characterization of novel stromal specific antibodies

    Institute of Scientific and Technical Information of China (English)


    Rheumatoid synovial fibroblasts were used as an immunogen to produce monoclonal antibodies selected for their reactivity with stromal cell antigens. Mice were immunised with low passage whole cell preparations and the subsequent hybridomas were screened by immunohistochemistry on rheumatoid synovium and tonsil sections. The aim was to identify those antibodies that recognised antigens that were restricted to stromal cells and were not expressed on CD45 positive leucocytes. A significant number of antibodies detected antigen that identified endothelial cells. These antibodies were further characterised to determine whether the vessels identified by these antibodies were vascular or lymphatic.From five fusions clones were identified with predominant reactivity with: 1) fibroblasts and endothelial cells; or 2)broad stromal elements (fibroblast, endothelium, epithelium, follicular dendritic cells). A fibroblast-specific antibody that did not also identify vessels was not generated. Examples of each reactivity pattern are discussed.

  19. Updates: Routine screening for antibodies to human immunodeficiency virus, type 1 (HIV-1), civilian applicants for U.S. military service and U.S. Armed Forces, active and reserve components. (United States)


    During routine testing of civilian applicants for U.S. military service, the overall seroprevalence of antibodies to HIV-1 in 2011 was the second lowest of any year since 1990. Among members of the active components of the U.S. Army, HIV-1 seroprevalences were higher during 2008 to 2011 than in recent prior years. Among members of the active components of the U.S. Air Force, Navy and Marine Corps, the Marine Corps Reserve, and the Army National Guard, HIV-1 seroprevalences have slightly declined or remained relatively stable for at least ten years. In the reserve components of most service branches, it is difficult to discern long-term trends because of instability of seroprevalences in the relatively small numbers of reserve component members tested each year. Monitoring of HIV-1 seroprevalences can help target and focus prevention initiatives. The recent repeal of the Don't Ask Don't Tell policy has created opportunities for prevention messages targeted to men who have sex with men.

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

    Directory of Open Access Journals (Sweden)

    Mehrangiz Rajaii


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

  1. Discovery of functional antibodies targeting ion channels. (United States)

    Wilkinson, Trevor C I; Gardener, Matthew J; Williams, Wendy A


    Ion channels play critical roles in physiology and disease by modulation of cellular functions such as electrical excitability, secretion, cell migration, and gene transcription. Ion channels represent an important target class for drug discovery that has been largely addressed, to date, using small-molecule approaches. A significant opportunity exists to target these channels with antibodies and alternative formats of biologics. Antibodies display high specificity and affinity for their target antigen, and they have the potential to target ion channels very selectively. Nevertheless, isolating antibodies to this target class is challenging due to the difficulties in expression and purification of ion channels in a format suitable for antibody drug discovery in addition to the complexity of screening for function. In this article, we will review the current state of ion channel biologics discovery and the progress that has been made. We will also highlight the challenges in isolating functional antibodies to these targets and how these challenges may be addressed. Finally, we also illustrate successful approaches to isolating functional monoclonal antibodies targeting ion channels by way of a number of case studies drawn from recent publications.

  2. Study of non-organ-specific antibodies in children with Genotype 4 chronic Hepatitis C

    Directory of Open Access Journals (Sweden)

    Mohammed E Hamed


    Full Text Available Background/Aim: Adult studies established a relationship between hepatitis C virus (HCV infection and the presence of non-organ-specific antibodies (NOSAs. Most studies were carried out on genotypes 1 and 2. Only a few studies addressed that issue in pediatrics. No studies have been carried out on autoimmunity and genotype 4 in children. We aim to investigate NOSAs in 80 Egyptian children with chronic HCV infection along with studying the underlying genotype of HCV, and correlating autoimmunity with the epidemiological, clinical, biochemical, and virological features. Materials and Methods: HCV-RNA was assayed by the polymerase chain reaction and viral genotypes were determined. NOSAs were measured and liver biopsies were taken for histopathological examination. Results:Genotype 4 was the only detected genotype in the included 80 patients. Anti-smooth muscle antibodies (ASMA were the only detected antibodies in 32 (40% patients, always with V specificity (vessels only at titers ranging from 1:20 and 1:160. Anti-nuclear antibodies (ANA and liver-kidney microsomal antibodies-1 (LKMA-1 were not detected in any of our patients. Epidemiologic and clinical features did not significantly differ between autoantibody-positive and -negative patients. Among biochemical features, significantly high levels of total bilirubin, albumin, immunoglobulins, alkaline phosphatase, and gamma-glutamyl transpeptidase were found in the antibody-positive group. Conclusion: Genotype 4 HCV is the prevailing genotype in Egyptian children with chronic HCV infection. A consistent proportion of these children with chronic HCV infection circulate non-organ-specific autoantibodies. The prevalence of ASMA and the absence of ANA and LKMA-1 might be related to the unique situation in Egypt with unique prevalence of genotype 4. More studies are warranted on larger pediatric population to validate these findings.

  3. [VGKC-complex antibodies]. (United States)

    Watanabe, Osamu


    Various antibodies are associated with voltage-gated potassium channels (VGKCs). Representative antibodies to VGKCs were first identified by radioimmunoassays using radioisotope-labeled alpha-dendrotoxin-VGKCs solubilized from rabbit brain. These antibodies were detected only in a proportion of patients with acquired neuromyotonia (Isaacs' syndrome). VGKC antibodies were also detected in patients with Morvan's syndrome and in those with a form of autoimmune limbic encephalitis. Recent studies indicated that the "VGKC" antibodies are mainly directed toward associated proteins (for example LGI-1 and CASPR-2) that complex with the VGKCs themselves. The "VGKC" antibodies are now commonly known as VGKC-complex antibodies. In general, LGI-1 antibodies are most commonly detected in patients with limbic encephalitis with syndrome of inappropriate secretion of antidiuretic hormone. CASPR-2 antibodies are present in the majority of patients with Morvan's syndrome. These patients develop combinations of CNS symptoms, autonomic dysfunction, and peripheral nerve hyperexcitability. Furthermore, VGKC-complex antibodies are tightly associated with chronic idiopathic pain. Hyperexcitability of nociceptive pathways has also been implicated. These antibodies may be detected in sera of some patients with neurodegenerative diseases (for example, amyotrophic lateral sclerosis and Creutzfeldt-Jakob disease).

  4. Construction of human naive phage antibody library and primary screening of the gab antibodies against gp96%人天然噬菌体抗体库的构建及抗gp96抗体的初步筛选

    Institute of Scientific and Technical Information of China (English)

    马小兵; 畅继武; 李成文; 李慧忠; 王馨


    Objective To construct a naive human Fab fragment phage display library,from which the anti-gp96 antibodies may be panned by the gp96 purified from the tissue of urothelial carcinoma in the urinary bladder and provide a basis to new therapy for the malignant tumors.Methods Peripheral blood lymphocytes were isolated from 800 ml of blood,which was obtained from four healthy blood donors.The heavy chain Fd and light chain cDNA synthesized from the total RNA of lympbocytes were amplified by PCR with variable regions 5' and 3' primers of heavy and light chain, and the amplification products were ligated into the phagemid vector pComb3, then the ligated sample was transformed into competent E.coli XL1-Blue by electroperation.The transformed cells were infected with VCSM13 helper phage to yield recombinant phage antibody Fabs.The phagemids abstracted from amplified E.coli were cut with endonucleases such as Sac Ⅰ,Xba Ⅰ,Xho Ⅰ and Spe Ⅰ, and both the phage antibody Fabs and phage-raids abstracted from amplified E.coil were amplified by PCR to monitor the insertion of the genes of light chain or heavy chain Fd fragment.The gp96 purified from the urothelial carcinoma tissue of the bladder by affinity chromatog-raphy on eoncanavalin-A sephnrose and DEAE-sephnrose ion exchange chromatography were utilized as antigens to process three rounds of panning to the original Fab antibody library.Results The quantity of total RNA and cDNA were qualified.By combination of light chain and heavy chain genes, an antibody library containing 6.6×106 clones was obtained, and both the cutting of enzymes and PCR showed that there were the genes of light chain or heavy chain Fd fragment in the phagemids.The gp96 protein was obtained from urothelial carcinoma tissue in the urinary bladder.After having been panned by gp96, the original antibody library gained enrichment by 68 times.Conclusion Utilizing the technology of phage surface display, specific antibody can be gained from the human

  5. 酶联免疫吸附试验检测单纯疱疹病毒抗体在生殖器疱疹筛查中的应用%Application of ELISA detection of herpes simplex virus antibodies in genital herpes screening

    Institute of Scientific and Technical Information of China (English)

    徐雪峰; 皮治兵


    目的:探讨酶联免疫吸附试验检测单纯疱疹病毒(HSV)抗体对生殖器疱疹筛查的意义。方法:选取50例2010年2月至2013年10月来我院治疗疑似生殖器疱疹患者,将其作为试验组,使用酶联免疫吸附试验进行HSV抗体检测,另外选择50例健康体检人员血清作为对照组。结果:在50例疑似生殖器疱疹患者HSV-I IgM,HSV-I IgG的检出率依次为14%,32%,HSV-ⅡIgM检出率为20%,HSV-ⅡIgG的检出率为34%;在对照组中HSV-I IgM,HSV-I IgG的检出率依次为0.8%,14%,HSV-ⅡIgM检出率为10%,HSV-ⅡIgG的检出率为24%,两组患者HSV抗体检测阳性率有显著的不同,二者明显的差异具有统计学意义(P<0.05)。结论:疑似生殖器疱疹患者单纯疱疹病毒抗体检出率高,酶联免疫检测HSV抗原的方法可直接检测出泌尿生殖道及皮损中HSV病原体,在生殖器疱疹筛查中具有一定的优势。%Objectives:To investigate the application of ELISA detection of herpes simplex virus (HSV) antibody in screening genital herpes.Method:50 patients,from February 201 0 to October 201 3,in our hospital with suspected genital herpes were selected as the test group,using enzyme-linked immunosorbent assay to detect HSV antibodies.In addition,50 healthy people were selected as control group.Results:In the 50 cases of test group,the detection rates of HSV-I IgMand HSV-I IgG were 1 4%and 32%respectively.HSV-ⅡIgMdetec-tion rate was 20%.HSV-ⅡIgG detection rate was 34%.In the control group,HSV-I IgM and HSV-I IgG de-tection rates were 0.8% and 1 4%,respectively.HSV-Ⅱ IgM detection rate was 1 0%.HSV-Ⅱ IgG detection rate was 24%.The difference of the HSV antibody positive rates in the two groups were statistically significant (P<0. 05 ).Conclusion:The detection rate of simplex virus antibody in patients with suspected genital herpes is high. ELISA method can directly detect HSV antigen

  6. Combinative application CD4+ CD25+ regulatory T cells test and irregular antibody screening in the diagnosis of invalid clinical transfusion%联合检测CD4+ CD25+调节性T细胞和血清不规则抗体在红细胞输注无效诊断的应用研究

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate whether the T cells (CD4+ CD25+) levels and the irregular antibodies screening cloud improve the diagnosis of invalid red blood cells transfusion.Methods Thirty-one patients with red blood cell invalid transfusion in the People's Hospital of Yubei District of Chongqing were selected.Flow cytometry was used to detect the changes of CD4+CD25+ regulatory T cells.Standard cells Ⅰ,Ⅱ,Ⅲ were used to screen on irregular antibodies in red blood cells (RBC).Results The rate of invalid RBC transfusion was 7.52% (31/412) in Yubei District.The incidence rate of medical diseases was 77.42% (24/31),much higher than surgical disease (22.58% (7/31,P =0.002),and the major departments were oncology department,hematological department and infectious department.The CD4+ CD25+ regulatory T cells were decrease from (22.18±1.58) % to (16.57±1.77) %(P=0.023).Conclusion Combin CD4+CD25+ regulatory T cells test and irregular antibody screening can help prevent and reduce the invalid transfusionof red blood cells.%目的 通过联合检测红细胞输注前后调节性T细胞(CD4+ CD25+)的变化和不规则抗体存在情况,探讨其是否有利于提高患者红细胞输注无效的检出率.方法 选取渝北区人民医院红细胞输注无效患者31例,采用流式细胞术检测输血前后CD4+ CD25+调节性T细胞变化,采用标准筛查细胞Ⅰ、Ⅱ、Ⅲ检测血清低效价、低亲和力不规则抗体.结果 渝北区红细胞输注无效发生率为7.52%(31/412),内科系统患者红细胞输注无效构成比为77.42%(24/31),远大于外科系统患者的22.58%(7/31) (P=0.002),红细胞输注无效发生以肿瘤科、血液内科、感染科为主;红细胞输注无效患者CD4+CD25+调节性T细胞表达由(22.18±1.58)%下降至(16.57±1.77)% (P=0.023).结论 联合检测CD4+ CD25+T细胞的变化和不规则抗体表达,有利于提高红细胞输注无效的检出率,预防和减少红细胞输注无效.

  7. Drug-induced hepatitis superimposed on the presence of anti-SLA antibody: a case report

    Directory of Open Access Journals (Sweden)

    Etxagibel Aitziber


    Full Text Available Abstract Introduction Autoimmune hepatitis is a necroinflammatory disorder of unknown etiology characterized by the presence of circulating antibodies, hypergammaglobulinemia, and response to immunosuppression. It has the histological features of chronic hepatitis. The onset is usually insidious, but in some patients the presentation may be acute and occasionally severe. Certain drugs can induce chronic hepatitis mimicking autoimmune hepatitis. Different autoantibodies have been associated with this process but they are not detectable after drug withdrawal and clinical resolution. Case presentation We describe a case of drug-induced acute hepatitis associated with antinuclear, antisoluble liver-pancreas and anti-smooth muscle autoantibodies in a 66-year-old woman. Abnormal clinical and biochemical parameters resolved after drug withdrawal, but six months later anti-soluble liver-pancreas antibodies remained positive and liver biopsy showed chronic hepatitis and septal fibrosis. Furthermore, our patient has a HLA genotype associated with autoimmune hepatitis. Conclusion Patient follow-up will disclose whether our patient suffers from an autoimmune disease and if the presence of anti-soluble liver antigens could precede the development of an autoimmune hepatitis, as the presence of antimitochondrial antibodies can precede primary biliary cirrhosis.

  8. Perinuclear antineutrophilic cytoplasmic antibody-positive cutaneous polyarteritis nodosa associated with minocycline therapy for acne vulgaris. (United States)

    Schaffer, J V; Davidson, D M; McNiff, J M; Bolognia, J L


    Minocycline is an oral antibiotic widely used for the long-term treatment of acne vulgaris. Unusual side effects of this medication include two overlapping autoimmune syndromes: drug-induced lupus and autoimmune hepatitis. In addition, in a few patients livedo reticularis or subcutaneous nodules have developed in association with arthritis and serum perinuclear antineutrophil cytoplasmic antibodies (P-ANCA) during long-term minocycline therapy. We report the cases of two young women receiving long-term minocycline therapy (>3 years) in whom P-ANCA-positive cutaneous polyarteritis nodosa developed. Both patients presented with a violaceous reticulated pattern on the lower extremities. Histologic examination of biopsy specimens from a reticulated area and a subcutaneous nodule showed necrotizing vasculitis of medium-sized arteries in the deep dermis, consistent with the diagnosis of polyarteritis nodosa. The cutaneous lesions rapidly resolved on discontinuation of minocycline and initiation of prednisone therapy. A high index of suspicion and testing for antineutrophil cytoplasmic antibody in addition to the standard antinuclear antibody panel can facilitate diagnosis of minocycline-related autoimmune disorders.

  9. Non-covalent carriage of anticancer agents by humanized antibody trastuzumab. (United States)

    Yadav, Arpita; Sharma, Sweta; Yadav, Veejendra Kumar


    This article explores the internalization and non-covalent carriage of small molecule anticancer agents like vinca alkaloids by humanized monoclonal antibody trastuzumab. Such carriage is marked by significant reduction in side effects and increased therapeutic value of these anticancer agents. This study is coherent with few clinical observations of enhanced efficiency of these anticancer agents when co-administered with therapeutic antibodies. This study will also serve as the foundation for screening a database of anticancer agents for possible compounds that may be co-delivered alongwith the antibody. Based on this study vincristine conformation inside antibody and its charge environment may be used as descriptors for screening purposes.


    Directory of Open Access Journals (Sweden)

    Shahnaz Rafiei Tehrani


    Full Text Available Fifty-seven patients suffering from systemic lupus erythematosus with different clinical features were studied from various aspects of immunological abnormalities, including anti-nuclear antibodies; anti-DNA; anti-Sm (Smith's antigen; and, other extractable nuclear antigens."nThe tests employed were microhemagglutination, counter immunoelecterphoresis, and radio immunoassay (RIA."nThe comparison between the two techniques for detectiong anti-DNA titers showed a significant correlation between them. There was also a significant positive correlation between the presence of anti-DNA and lupus nephritis (P>O.05. However, lupus nephritis has a negative association with anti-Sm."nWe will discuss anti-Sm in SLE patients that may have an inhibitory effect on developing the lupus nephritis. The association of anti-Sm in SLE may help the physicians to prognose the disease and manipulate the treatment.

  11. The association of anti-annexin1 antibodies with the occurrence of skin lesions in systemic lupus erythematosus. (United States)

    Meng, Z; Shi, Z-R; Tan, G-Z; Yin, J; Wu, J; Mi, X-B; Wang, L


    Anti-annexin1 antibodies are associated with the subtypes of cutaneous lupus and are elevated in systemic lupus erythematosus (SLE) patients. In this study, we investigated the correlation of this antibody with the incidence of SLE skin lesions. The presence of anti-annexin1-IgG and-IgM determined by Western blot was no different among healthy controls and SLE patients with and without skin lesions. Serum levels of anti-annexin1-IgG and -IgM measured by enzyme-linked immunosorbent assay were comparable between patients with and without skin lesions, whereas anti-annexin1-IgM was lower in SLE patients than in healthy controls. Annexin1 was abundantly detected in each epidermal layer in lupus lesional skin. Additionally, anti-annexin1-IgG was higher in SLE patients with arthritis and negatively correlated with white blood cells (WBC). Anti-annexin1-IgM was higher in patients with antinuclear antibody (ANA)-positive sera, and was positively related to hemoglobin and total serum IgM. Collectively, anti-annexin1 antibodies are not related to the incidence of skin lesions in SLE, and annexin1 abundantly distributes in epidermis in lesional skin.

  12. 单纯疱疹病毒检测在宫颈疾病中的价值%Study on significance of high-risk type HSV antibody testing in screening of cervical diseases

    Institute of Scientific and Technical Information of China (English)

    赵杨; 李晓平


    目的 探讨单纯疱疹病毒(HSV)感染在宫颈疾病中的临床特点以及价值.方法 采用酶联免疫吸附试验(ELISA)对患者血清标本进行单纯疱疹(HSV-1及HSV-2)特异性病毒抗体(IgM)检测.结果 1095例患有子宫颈糜烂、滴虫性和霉菌性阴道炎及淋病患者血清标本HSV-IgM阳性检出率分别为32.5%、23.4%、15.7%、22.8%,除淋病组与滴虫组、霉菌组与淋病组比较差异无统计学意义外,其他几组都有HSV感染率均有显著性差异.378例宫颈糜烂患者根据程度不同HSV阳性率分别为16.6%、33.6%、43.5%,三组之间有显著性差异,且随糜烂程度加重HSV阳性率增高.86例不同病理分型患者HSV阳性检出率分别为26.1%、40%、50、62.5%,HSV阳性检出率随病理分级严重程度呈趋势增高,但无显著性差异.结论 HSV感染可以与上述疾病单发或混合感染,HSV是宫颈疾病发生、进展的关键因素之一;HSV感染的检测对妇女生殖疱器疹的诊断及治疗,对围产医学和优生优育等工作均有重大作用.%Objective To evaluate clinical features and significance of herpes simplex virus infection in patients with cervical diseases. Methods Serum levels of anti - HSV -1 and anti - HSV - 2 specific IgM antibodies in patients were detected by using enzyme - linked immu-nosorbent assay ( ELISA ). Results The positive rates of serum anti - HSV - IgM in 1095 patients with cervical erosion, gonorrhea, trichomoniasis or fungal vaginitis were 32. 5% , 22. 8% , 23.9% and 15. 6% respectively. There was significant difference in HSV infection rates among patients in all groups except gonorrhea group with trichomoniasis group and fungus infection group with gonorrhea group. The positive rates of anti - HSV -IgM in 378 patients with different degrees of cervical erosion were 16.6% , 33. 6% and 43.5% respectively, and there was significant difference among these three groups. The positive rate of anti - HSV IgM was raised following the

  13. Pyridoxal-5'-phosphate-dependent catalytic antibodies. (United States)

    Gramatikova, Svetlana; Mouratou, Barbara; Stetefeld, Jörg; Mehta, Perdeep K; Christen, Philipp


    Strategies for expanding the catalytic scope of antibodies include the incorporation of inorganic or organic cofactors into their binding sites. An obvious choice is pyridoxal-5'-phosphate (PLP), which is probably the most versatile organic cofactor of enzymes. Monoclonal antibodies against the hapten N(alpha)-(5'-phosphopyridoxyl)-L-lysine, a stable analog of the covalent coenzyme-substrate adducts were screened by a competition ELISA for binding of the PLP-amino acid Schiff base adduct. The Schiff base with its C4'-N alpha double bond is, in contrast to the hapten, a planar compound and is an obligatory intermediate in all PLP-dependent reactions of amino acids. This highly discriminating screening step eliminated all but 5 of 24 hapten-binding antibodies. The five remaining antibodies were tested for catalysis of the PLP-dependent alpha,beta-elimination reaction of beta-chloroalanine. Antibody 15A9 complied with this selection criterion and catalyzed in addition the cofactor-dependent transamination reaction of hydrophobic D-amino acids and oxo acids (k(cat)'=0.42 min(-1) with D-alanine at 25 degrees C). Homology modeling together with alanine scanning yielded a 3D model of Fab 15A9. The striking analogy between antibody 15A9 and PLP-dependent enzymes includes the following features: (1) The binding sites accommodate the planar coenzyme-amino acid adduct. (2) The bond at C alpha to be broken lies together with the C alpha-N bond in a plane orthogonal to the plane of coenzyme and imine bond. (3) The alpha-carboxylate group of the substrate is bound by an arginine residue. (4) The coenzyme-substrate adduct assumes a cisoid conformation. (5) PLP markedly contributes to catalytic efficiency, being a 10(4) times more efficient amino group acceptor than pyruvate. The protein moiety, however, ensures reaction as well as substrate specificity, and further accelerates the reaction (in 15A9 k(cat (Ab x PLP))'/k(cat (PLP))'=5 x 10(3)). The analogies of antibody 15A9 with

  14. Rapid isolation of antibody from a synthetic human antibody library by repeated fluorescence-activated cell sorting (FACS.

    Directory of Open Access Journals (Sweden)

    Sung Sun Yim

    Full Text Available Antibodies and their derivatives are the most important agents in therapeutics and diagnostics. Even after the significant progress in the technology for antibody screening from huge libraries, it takes a long time to isolate an antibody, which prevents a prompt action against the spread of a disease. Here, we report a new strategy for isolating desired antibodies from a combinatorial library in one day by repeated fluorescence-activated cell sorting (FACS. First, we constructed a library of synthetic human antibody in which single-chain variable fragment (scFv was expressed in the periplasm of Escherichia coli. After labeling the cells with fluorescent antigen probes, the highly fluorescent cells were sorted by using a high-speed cell sorter, and these cells were reused without regeneration in the next round of sorting. After repeating this sorting, the positive clones were completely enriched in several hours. Thus, we screened the library against three viral antigens, including the H1N1 influenza virus, Hepatitis B virus, and Foot-and-mouth disease virus. Finally, the potential antibody candidates, which show K(D values between 10 and 100 nM against the target antigens, could be successfully isolated even though the library was relatively small (∼ 10(6. These results show that repeated FACS screening without regeneration of the sorted cells can be a powerful method when a rapid response to a spreading disease is required.

  15. High throughput production of mouse monoclonal antibodies using antigen microarrays

    DEFF Research Database (Denmark)

    De Masi, Federico; Chiarella, P.; Wilhelm, H.;


    Recent advances in proteomics research underscore the increasing need for high-affinity monoclonal antibodies, which are still generated with lengthy, low-throughput antibody production techniques. Here we present a semi-automated, high-throughput method of hybridoma generation and identification....... Monoclonal antibodies were raised to different targets in single batch runs of 6-10 wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen-coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated...

  16. Expression of recombinant antibodies. (United States)

    Frenzel, André; Hust, Michael; Schirrmann, Thomas


    Recombinant antibodies are highly specific detection probes in research, diagnostics, and have emerged over the last two decades as the fastest growing class of therapeutic proteins. Antibody generation has been dramatically accelerated by in vitro selection systems, particularly phage display. An increasing variety of recombinant production systems have been developed, ranging from Gram-negative and positive bacteria, yeasts and filamentous fungi, insect cell lines, mammalian cells to transgenic plants and animals. Currently, almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with "human-like" post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. In this review, we focus on current antibody production systems including their usability for different applications.

  17. Therapeutic Recombinant Monoclonal Antibodies (United States)

    Bakhtiar, Ray


    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  18. Recombinant renewable polyclonal antibodies. (United States)

    Ferrara, Fortunato; D'Angelo, Sara; Gaiotto, Tiziano; Naranjo, Leslie; Tian, Hongzhao; Gräslund, Susanne; Dobrovetsky, Elena; Hraber, Peter; Lund-Johansen, Fridtjof; Saragozza, Silvia; Sblattero, Daniele; Kiss, Csaba; Bradbury, Andrew R M


    Only a small fraction of the antibodies in a traditional polyclonal antibody mixture recognize the target of interest, frequently resulting in undesirable polyreactivity. Here, we show that high-quality recombinant polyclonals, in which hundreds of different antibodies are all directed toward a target of interest, can be easily generated in vitro by combining phage and yeast display. We show that, unlike traditional polyclonals, which are limited resources, recombinant polyclonal antibodies can be amplified over one hundred million-fold without losing representation or functionality. Our protocol was tested on 9 different targets to demonstrate how the strategy allows the selective amplification of antibodies directed toward desirable target specific epitopes, such as those found in one protein but not a closely related one, and the elimination of antibodies recognizing common epitopes, without significant loss of diversity. These recombinant renewable polyclonal antibodies are usable in different assays, and can be generated in high throughput. This approach could potentially be used to develop highly specific recombinant renewable antibodies against all human gene products.

  19. Breast cancer screening (United States)

    Mammogram - breast cancer screening; Breast exam - breast cancer screening; MRI - breast cancer screening ... performed to screen women to detect early breast cancer when it is more likely to be cured. ...

  20. Development and evaluation of an enzyme-labeled antibody test for the rapid detection of hog cholera antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Saunders, G.C.


    A rapid enzyme-labeled antibody (ELA) microtechnique for the screening of swine for hog cholera antibodies was developed and evaluated with a blind study, using a 640-sample hog cholera serum bank. The total time to run a group of 22 samples was approximately 1 hour. The ELA test results correlated >99% with hog cholera serum-neutralization test results on the same serums. Test results also indicated that the ELA test shares with the hog cholera serum-neutralization test the problem of cross reactions between the antibodies of hog cholera and bovine viral diarrhea.

  1. Anti-insulin antibody test (United States)

    Insulin antibodies - serum; Insulin Ab test; Insulin resistance - insulin antibodies; Diabetes - insulin antibodies ... You appear to have an allergic response to insulin Insulin no longer seems to control your diabetes

  2. Profiling anti-cyclic citrullinated peptide antibodies in patients with juvenile idiopathic arthritis

    Directory of Open Access Journals (Sweden)

    Tebo Anne E


    Full Text Available Abstract Background Anti-citrullinated protein/peptide antibodies (ACPA, have high specificity for rheumatoid arthritis (RA. Some children with juvenile idiopathic arthritis (JIA, phenotypically resemble RA and test positive for rheumatoid factor (RF a characteristic biomarker of RA. We investigated the prevalence of ACPA and its relationship to other serologic markers associated with RA in a well-characterized JIA cohort. Methods Cases were 334 children with JIA, 30 of whom had RF + polyarticular JIA. Sera from all cases and 50 healthy pediatric controls were investigated by ELISA at a single time point for anti-cyclic citrullinated peptide (anti-CCP IgG, RF IgM, IgA and IgG, anti-RA33 IgG, and antinuclear antibodies (ANA. Comparisons between cases and controls were made using Chi-square or Fisher exact tests and T-tests. Results The prevalence of RF was 8% among controls, and 12% among cases (ns. The prevalence of ACPA was 2% in controls and 14.3% in cases (OR 8.2, p Conclusions ACPAs are detectable in 14% of children with JIA. Children with positive ACPA but negative RF are frequent, and may define a distinct subset of children with JIA. ACPA testing should be included in the classification of JIA.

  3. Monoclonal antibody "gold rush". (United States)

    Maggon, Krishan


    The market, sales and regulatory approval of new human medicines, during the past few years, indicates increasing number and share of new biologics and emergence of new multibillion dollar molecules. The global sale of monoclonal antibodies in 2006 were $20.6 billion. Remicade had annual sales gain of $1 billion during the past 3 years and five brands had similar increase in 2006. Rituxan with 2006 sales of $4.7 billion was the best selling monoclonal antibody and biological product and the 6th among the top selling medicinal brand. It may be the first biologic and monoclonal antibody to reach $10 billion annual sales in the near future. The strong demand from cancer and arthritis patients has surpassed almost all commercial market research reports and sales forecast. Seven monoclonal antibody brands in 2006 had sales exceeding $1 billion. Humanized or fully human monoclonal antibodies with low immunogenicity, enhanced antigen binding and reduced cellular toxicity provide better clinical efficacy. The higher technical and clinical success rate, overcoming of technical hurdles in large scale manufacturing, low cost of market entry and IND filing, use of fully human and humanized monoclonal antibodies has attracted funds and resources towards R&D. Review of industry research pipeline and sales data during the past 3 years indicate a real paradigm shift in industrial R&D from pharmaceutical to biologics and monoclonal antibodies. The antibody bandwagon has been joined by 200 companies with hundreds of new projects and targets and has attracted billions of dollars in R&D investment, acquisitions and licensing deals leading to the current Monoclonal Antibody Gold Rush.

  4. Antiphospholipid antibody syndrome. (United States)

    Kutteh, William H; Hinote, Candace D


    Antiphospholipid antibodies (aPLs) are acquired antibodies directed against negatively charged phospholipids. Obstetric antiphospholipid antibody syndrome (APS) is diagnosed in the presence of certain clinical features in conjunction with positive laboratory findings. Obstetric APS is one of the most commonly identified causes of recurrent pregnancy loss. Thus, obstetric APS is distinguished from APS in other organ systems where the most common manifestation is thrombosis. Several pathophysiologic mechanisms of action of aPLs have been described. This article discusses the diagnostic and obstetric challenges of obstetric APS, proposed pathophysiologic mechanisms of APS during pregnancy, and the management of women during and after pregnancy.

  5. Debye screening (United States)

    Brydges, David C.; Federbush, Paul


    The existence and exponential clustering of correlation functions for a classical coulomb system at low density or high temperature are proven using methods from constructive quantum field theory, the sine gordon transformation and the Glimm, Jaffe, Spencer expansion about mean field theory. This is a vindication of a belief of long standing among physicists, known as Debye screening. That is, because of special properties of the coulomb potential, the configurations of significant probability are those in which the long range parts of r -1 are mostly cancelled, leaving an effective exponentially decaying potential acting between charge clouds. This paper generalizes a previous paper of one of the authors in which these results were obtained for a special lattice system. The present treatment covers the continuous mechanics situation, with essentially arbitrary short range forces and charge species. Charge symmetry is not assumed.

  6. Squirrel monkey cytomegalovirus antibodies in free-ranging black howler monkeys (Alouatta caraya), Misiones, Argentina. (United States)

    Ferreyra, Hebe; Argibay, Hernan; Rinas, Miguel A; Uhart, Marcela


    Serum from four black howler monkeys (Alouatta caraya) was screened for antibodies to seven viruses by dot immunoassay. Cytomegalovirus antibodies were detected in three of four individuals and provide the first evidence of exposure by black howler monkeys to this virus.

  7. Triagem sorológica de familiares de pacientes com doença celíaca: anticorpos anti-endomísio, antitransglutaminase ou ambos? Serological screening of relatives of celiac disease patients: antiendomysium antibodies, anti-tissue transglutaminase or both?

    Directory of Open Access Journals (Sweden)

    Shirley Ramos da Rosa Utiyama


    -negativas. O impacto desse fato implica que tais familiares deixarão de ser submetidos a biopsia intestinal para confirmação do diagnóstico da doença, e conseqüentemente, ao tratamento adequado e precoce.BACKGROUND: Celiac disease is the most common intestinal disorder of caucasian populations and presents a prevalence of 8% to 18% between the relatives of patients. The anti-endomysial (IgA-EmA and anti-tissue transglutaminase antibodies (IgA-tTG have represented an important non invasive and sensitivity method of screening and diagnosis of celiac disease in risk groups and populations. AIM: To investigate the prevalence of IgA-EmA and IgA-tTG antibodies in relatives of celiac patients and verify the degree of concordance between them. METHODS: One hundred and seventy seven relatives of celiac patients (76(feminino; 101(masculino; 2-79 years and 93 healthy individuals were evaluated (34(feminino; 59(masculino; 2-71 years. IgA-EmA were detected by indirect immunofluorescence, with human umbilical cord as substrate, while anti-IgA-tTG titers were measured by enzyme-linked immunosorbent assay (ELISA, using commercial kit. RESULTS: Total positivity to antibodies in relatives of celiac patients was of 21% (37/177, and showed significant difference compared to control group (0%; 0/93. Twelve percent (21/177 of celiac disease relatives were positive to IgA-EmA, 13.56% (24/177 to IgA-tTG, and 4.52% (8/177 to both assays simultaneously. The concordance between both methods was 83.6% (148/177 and the discordance was 16.4% (29/177, with a positive and significant correlation (r = 0.435. Among the concordant results, 79.1% (140/177 were negative and 4.52% (8/177 were positive to both antibodies. Among the discordant results, 7.34% (13/177 were positive to IgA-EmA and negative to IgA-tTG, while 9.04% (16/177 were negative to IgA- EmA and positive to IgA-tTG. CONCLUSION: Although the high positivity to IgA-EmA and IgA-tTG emphasizes the importance of the serological screening in

  8. Research of serum pepsinogen and Helicobacter pylori-IgG antibody in the screening of atrophic gastritis%血清胃蛋白酶原及幽门螺杆菌-IgG抗体在查萎缩性胃炎中的作用研究

    Institute of Scientific and Technical Information of China (English)



    Objective To analyze the use of serum pepsinogen(PG) and Helicobacter pylori(Hp)-IgG antibody in the screening of atrophic gastritis.Methods A total of 222 patients were divided into two groups according to the gastroscopy results:159 cases in observation group(atrophic gastritis) and 63 cases in control group(non-atrophic gastritis).Patients in observation group were divided into gastric antrum group(50cases),gastric body group (71 cases) and full stomach multifocal group (3 8 cases) according to shrinking parts.The serum PG Ⅰ and PG Ⅱ was determined by enzyme linked immunosorbent assay method,and PGR (PG Ⅰ / PG Ⅱ) was calculated.The serum Hp-IgG antibody was examined by qualitative analysis.Results The serum PG Ⅰ and PGR was (57.82 ± 23.15) μ g/L,4.41 ± 1.82 in observation group,which was lower than that in control group[(125.04 ± 29.36) μ g/L,10.10 ± 2.01],and there was significant difference (P <0.01).The positive rate of the serum Hp-IgG antibody was 86.16%(137/159) in observation group,which was higher than that in control group [53.97% (34/63)],and there was significant difference (P < 0.01).There was significant difference in the serum PG Ⅰ and PGR between gastric antrum group and gastric body group,full stomach multifocal group (P < 0.01).There was no significant difference in the serum PG Ⅰ and PGR among different atrophy degree in gastric antrum group(P> 0.05),but there was significant difference in the serum PG Ⅰ and PGR among different atrophy degree in gastric body group and full stomach multifocal group (P < 0.01).Conclusions The serum PG Ⅰ and PGR and Hp-IgG antibody is related to atrophic gastritis.They are helpful to the screening of atrophic gastritis.%目的 分析血清胃蛋白酶原(PG)及幽门螺杆菌(Hp)-IgG抗体在萎缩性胃炎筛查中的作用.方法 222例患者根据胃镜检查结果分为两组:观察组(萎缩性胃炎)159例及对照组(非萎缩性胃炎)63例.其中观

  9. Anti-cartilage antibody. (United States)

    Greenbury, C L; Skingle, J


    Antibody to cartilage has been demonstrated by indirect immunofluorescence on rat trachea in the serum of about 3% of 1126 patients with rheumatoid arthritis. Titres ranged from 1:20 to 1:640. The antibody was not found in 284 patients with primary or secondary osteoarthritis or in 1825 blood donors, nor, with the exception of two weak reactors, in 1314 paraplegic patients. In most cases the antibody appears to be specific for native type II collagen. Using this as an antigen in a haemagglutination test 94% of anti-cartilage sera were positive, whereas among 100 rheumatoid control sera there were only three weak positives. More than 80% of patients with antibody had some erosion of articular cartilage, but there was no correlation with age, sex, duration of disease, nor any recognisable clinical event or change.

  10. Antithyroid microsomal antibody (United States)

    ... to confirm the cause of thyroid problems, including Hashimoto thyroiditis . The test is also used to find ... positive test may be due to: Granulomatous thyroiditis Hashimoto thyroiditis High levels of these antibodies have also ...

  11. Serum herpes simplex antibodies (United States)

    ... 2. HSV-1 most often causes cold sores (oral herpes). HSV-2 causes genital herpes. How the Test ... whether a person has ever been infected with oral or genital herpes . It looks for antibodies to herpes simplex virus ...

  12. A novel assay for monitoring internalization of nanocarrier coupled antibodies

    Directory of Open Access Journals (Sweden)

    Pickering Edward M


    Full Text Available Abstract Background Discovery of tumor-selective antibodies or antibody fragments is a promising approach for delivering therapeutic agents to antigen over-expressing cancers. Therefore it is important to develop methods for the identification of target- and function specific antibodies for effective drug delivery. Here we describe a highly selective and sensitive method for characterizing the internalizing potential of multivalently displayed antibodies or ligands conjugated to liposomes into tumor cells. The assay requires minute amounts of histidine-tagged ligand and relies on the non-covalent coupling of these antibodies to fluorescent liposomes containing a metal ion-chelating lipid. Following incubation of cells with antibody-conjugated liposomes, surface bound liposomes are gently removed and the remaining internalized liposomes are quantitated based on fluorescence in a high throughput manner. We have termed this methodology "Chelated Ligand Internalization Assay", or CLIA. Results The specificity of the assay was demonstrated with different antibodies to the ErbB-2 and EGF receptors. Antibody-uptake correlated with receptor expression levels in tumor cell lines with a range of receptor expression. Furthermore, Ni-NTA liposomes containing doxorubicin were used to screen for the ability of antibodies to confer target-specific cytotoxicity. Using an anti-ErbB2 single chain Fv (scFv (F5 antibody, cytotoxicity could be conferred to ErbB2-overexpressing cells; however, a poly(ethylene glycol-linked lipid (DSPE-PEG-NTA-Ni was necessary to allow for efficient loading of the drug and to reduce nonspecific drug leakage during the course of the assay. Conclusion The CLIA method we describe here represents a rapid, sensitive and robust assay for the identification and characterization of tumor-specific antibodies capable of high drug-delivery efficiency when conjugated to liposomal nanocarriers.

  13. Developing recombinant antibodies for biomarker detection

    Energy Technology Data Exchange (ETDEWEB)

    Baird, Cheryl L.; Fischer, Christopher J.; Pefaur, Noah B.; Miller, Keith D.; Kagen, Jacob; Srivastava, Sudhir; Rodland, Karin D.


    Monoclonal antibodies (mAbs) have an essential role in biomarker validation and diagnostic assays. A barrier to pursuing these applications is the reliance on immunization and hybridomas to produce mAbs, which is time-consuming and may not yield the desired mAb. We recommend a process flow for affinity reagent production that utilizes combinatorial protein display systems (eg, yeast surface display or phage display) rather than hybridomas. These systems link a selectable phenotype-binding conferred by an antibody fragment-with a means for recovering the encoding gene. Recombinant libraries obtained from immunizations can produce high-affinity antibodies (<10 nM) more quickly than other methods. Non-immune libraries provide an alternate route when immunizations are not possible, or when suitable mAbs are not recovered from an immune library. Directed molecular evolution (DME) is an integral part of optimizing mAbs obtained from combinatorial protein display, but can also be used on hybridoma-derived mAbs. Variants can easily be obtained and screened to increase the affinity of the parent mAb (affinity maturation). We discuss examples where DME has been used to tailor affinity reagents to specific applications. Combinatorial protein display also provides an accessible method for identifying antibody pairs, which are necessary for sandwich-type diagnostic assays.

  14. Experiência da adoção do I e II Consensos Brasileiros de Fator Antinuclear por Imunofluorescência Indireta em Células HEp-2 em um hospital universitário Experience of an university hospital on the implementation of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells

    Directory of Open Access Journals (Sweden)

    Claudia Cilene Fernandes Correia Laurino


    Full Text Available OBJETIVO:Analisar a prevalência, padrões e títulos do Fator Antinuclear (FAN por imunofluorescência indireta (IFI em células HEp-2 em um hospital universitário após a adoção do I e II Consensos Nacional para Padronização dos Laudos de FAN em Células HEp-2. MÉTODO: Foi realizado um estudo transversal, em que foram revisados os laudos de FAN por IFI originários de solicitações encaminhadas ao Serviço de Patologia Clínica do Hospital de Clínicas de Porto Alegre (SPC/HCPA entre 2002 e 2005. RESULTADOS:Foram analisados 12.095 testes de FAN no período entre 2002 e 2005. As solicitações com resultado reagente foram de 2.577 (21,30%, com média anual de 644±233. Houve um aumento significativo na proporção de resultados reagentes posterior à adoção dos Consensos (p OBJECTIVE: To evaluate the prevalence of patterns and titers of antinuclear antibodies (ANA detected by indirect immunofluorescence (IIF technique on HEp-2 cells in a university hospital following the introduction of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells. METHODS:A transversal study was performed between 2002 and 2005 during which all ANA orders to Serviço de Hospital de Clínicas de Porto Alegre (SPC/HCPA and cognate results were reviewed. RESULTS:12.095 tests of ANA were revised. The number of positive results during this period was 2.577 (21.30%, annual mean 644 (SD: 233. A marked increase in the number of positive results was observed following the introduction of the Consensuses (p < 0.001. Rheumatology was the medical specialty which requested the highest number of ANA testing per patient although a significant decrease of these numbers was observed after the introduction of the Consensus in 2004 (p < 0,001. Nuclear fine speckled immunofluorescence labeling was the most frequently ANA pattern observed, 52.3% (453/866, and low ANA titers (1/80 and 1/160 more commonly detected (27.8% and 29.4%, respectively. CONCLUSION

  15. Heparin-Induced Thrombocytopenia Antibody Test (United States)

    ... Global Sites Search Help? Heparin-induced Thrombocytopenia PF4 Antibody Share this page: Was this page helpful? Also known as: Heparin-PF4 Antibody; HIT Antibody; HIT PF4 Antibody; Heparin Induced Antibody; ...

  16. Human single chain antibody to vascular endothelial growth factor: gene cloning, high-level expression, affinity maturation and bioactivity

    Institute of Scientific and Technical Information of China (English)

    阎锡蕴[1; 汤健[2; 吴小平[3; 王凤采[4; 李建生[5; 杨东玲[6


    Using antibody phage display technique, a human single chain antibody to vascular endothelial growth factor (VEGF) has been cloned. The antibody expression reached 45% of the total bacterial proteins. The purification and refolding of the antibody were completed in one step by using gel filtration chromatograph. ELISA analysis showed that the antibody not only specifically bound to human VEGF, but also competitively inhibited VEGF reacting with its receptors. In order to raise the affinity of the single chain antibody, its heavy chain variable region was randomly mutated using error-prone PCR and an antibody mutant library was constructed, from which a mutant with higher affinity was screened out. The three-dimensional structure and binding affinity of wild type and mutant antibody were compared. Our study provided a potential reagent for tumor angiogenic therapy and a significant model for antibody high-level expression and affinity maturation.

  17. Human single chain antibody to vascular endothelial growth factor:gene cloning, high-level expression, affinity maturation and bioactivity

    Institute of Scientific and Technical Information of China (English)


    Using antibody phage display technique,a human single chain antibody to vascular endothelial growth factor (VEGF) has been cloned.The antibody expression reached 45% of the total bacterial proteins.The purification and refolding of the antibody were completed in one step by using gel filtration chromatograph.ELISA analysis showed that the antibody not only specifically bound to human VEGF,but also competitively inhibited VEGF reacting with its receptors.In order to raise the affinity of the single chain antibody,its heavy chain variable region was randomly mutated using error-prone PCR and an antibody mutant library was constructed,from which a mutant with higher affinity was screened out.The three-dimensional structure and binding affinity of wild type and mutant antibody were compared.Our study provided a potential reagent for tumor angiogenic therapy and a significant model for antibody high-level expression and affinity maturation.

  18. [New antibodies in cancer treatment]. (United States)

    Pestalozzi, B C; Knuth, A


    Since the development of hybridoma technology in 1975 monoclonal antibodies with pre-defined specificity can be produced. Only twenty years later did it become possible to make therapeutic use of monoclonal antibodies in oncology. To this end it was necessary to attach the antigen-binding site of a mouse antibody onto the scaffold of a human antibody molecule. Such chimeric or "humanized" antibodies may be used in passive immunotherapy without eliciting an immune response. Rituximab and trastuzumab are such humanized antibodies. They are used today routinely in the treatment of malignant lymphoma and breast cancer, respectively. These antibodies are usually used in combination with conventional cytostatic anticancer drugs.

  19. Engineering antibodies for cancer therapy. (United States)

    Boder, Eric T; Jiang, Wei


    The advent of modern antibody engineering has led to numerous successes in the application of these proteins for cancer therapy in the 13 years since the first Food and Drug Administration approval, which has stimulated active interest in developing more and better drugs based on these molecules. A wide range of tools for discovering and engineering antibodies has been brought to bear on this challenge in the past two decades. Here, we summarize mechanisms of monoclonal antibody therapeutic activity, challenges to effective antibody-based treatment, existing technologies for antibody engineering, and current concepts for engineering new antibody formats and antibody alternatives as next generation biopharmaceuticals for cancer treatment.

  20. Specificities of monoclonal antibodies to domain I of alpha-gliadins. (United States)

    Ellis, H J; Doyle, A P; Wieser, H; Sturgess, R P; Ciclitira, P J


    Eight monoclonal antibodies were raised against a sequenced 54-amino-acid peptide of alpha-gliadin, which is thought to exacerbate coeliac disease. Five of the antibodies cross-reacted with coeliac non-toxic cereals. Two of eight of the antibodies bound specifically to coeliac toxic prolamins. These two antibodies cross-reacted with high molecular weight gliadins, which are closely related to alpha-gliadins and whose toxicity to patients with coeliac disease is unclear. The antibodies were screened by enzyme-linked immunosorbent assay against three amino-acid-sequenced peptides of alpha-gliadin with single amino-acid differences. Differential binding of antibody WC2 suggested that this antibody binds in the region of amino-acid residue 36, a proline residue, where there may be an antigenic beta-reverse turn. This proline residue forms part of a tetrapeptide motif, QQQP, which is thought to be present in all coeliac-active peptides.

  1. Generation and characterization of monoclonal antibodies specific to Coenzyme A

    Directory of Open Access Journals (Sweden)

    Malanchuk O. M.


    Full Text Available Aim. Generation of monoclonal antibodies specific to Coenzyme A. Methods. Hybridoma technique. KLH carrier protein conjugated with CoA was used for immunization. Screening of positive clones was performed with BSA conjugated to CoA. Results. Monoclonal antibody that specifically recognizes CoA and CoA derivatives, but not its precursors ATP and cysteine has been generated. Conclusion. In this study, we describe for the first time the production and characterization of monoclonal antibodies against CoA. The monoclonal antibody 1F10 was shown to recognize specifically CoA in Western blotting, ELISA and immunoprecipitation. These properties make this antiboby a particularly valuable reagent for elucidating CoA function in health and disease.

  2. Lung Cancer Screening (United States)

    ... Treatment Lung Cancer Prevention Lung Cancer Screening Research Lung Cancer Screening (PDQ®)–Patient Version What is screening? Go ... These are called diagnostic tests . General Information About Lung Cancer Key Points Lung cancer is a disease in ...

  3. Skin Cancer Screening (United States)

    ... Genetics of Skin Cancer Skin Cancer Screening Research Skin Cancer Screening (PDQ®)–Patient Version What is screening? ... These are called diagnostic tests . General Information About Skin Cancer Key Points Skin cancer is a disease ...

  4. Mental Health Screening Center (United States)

    ... Releases & Announcements Public Service Announcements Partnering with DBSA Mental Health Screening Center These online screening tools are not ... you have any concerns, see your doctor or mental health professional. Depression This screening form was developed from ...

  5. Testicular Cancer Screening (United States)

    ... Health Professional Testicular Cancer Treatment Testicular Cancer Screening Testicular Cancer Screening (PDQ®)–Patient Version What is screening? Go ... These are called diagnostic tests . General Information About Testicular Cancer Key Points Testicular cancer is a disease in ...

  6. Natural and Man-made Antibody Repertories for Antibody Discovery

    Directory of Open Access Journals (Sweden)

    Juan C eAlmagro


    Full Text Available Antibodies are the fastest-growing segment of the biologics market. The success of antibody-based drugs resides in their exquisite specificity, high potency, stability, solubility, safety and relatively inexpensive manufacturing process in comparison with other biologics. We outline here the structural studies and fundamental principles that define how antibodies interact with diverse targets. We also describe the antibody repertoires and affinity maturation mechanisms of human, mice and chickens, plus the use of novel single-domain antibodies in camelids and sharks. These species all utilize diverse evolutionary solutions to generate specific and high affinity antibodies and illustrate the plasticity of natural antibody repertoires. In addition, we discuss the multiple variations of man-made antibody repertoires designed and validated in the last two decades, which have served as tools to explore how the size, diversity and composition of a repertoire impact the antibody discovery process.

  7. Anticorpos antinucleossomo e síndrome antifosfolipídica: estudo observacional Antinucleosome antibodies and primary antiphospholipid syndrome: an observational study

    Directory of Open Access Journals (Sweden)

    Alexandre Wagner Silva de Souza


    Full Text Available OBJETIVO: Avaliar a associação entre a presença de anticorpos antinucleossomo (anti-NCS e a síndrome antifosfolipídica primária (SAFP e o posterior desenvolvimento de lúpus eritematoso sistêmico (LES. MATERIAIS E MÉTODOS: Trinta e seis mulheres com o diagnóstico de SAFP foram avaliadas prospectivamente para manifestações de doenças reumáticas autoimunes e para a presença de anticorpos antifosfolípides, anticorpos antinucleares e anti-NCS/cromatina. RESULTADOS: Após um período médio de seguimento de 45,7 meses, anticorpos anti-NCS/cromatina foram detectados em apenas uma paciente (2,8%, que desenvolveu manifestações de LES tais como poliartrite, linfopenia, neurite óptica, lesões compatíveis com esclerose múltipla em substância branca cerebral e perfil de autoanticorpos altamente sugestivo de LES. CONCLUSÃO: A frequência de anticorpos anti-NCS/cromatina é baixa em pacientes com SAFP, e sua presença pode associar-se ao desenvolvimento de manifestações de LES.OBJECTIVE: To study the association of anti-nucleosome (anti-NCS antibodies in primary antiphospholipid syndrome (APS and the development of systemic lupus erythematosus (SLE during follow-up. MATERIALS AND METHODS: Thirty-six women with primary APS were evaluated prospectively for clinical features of systemic autoimmune diseases and for the presence of antiphospholipid antibodies, antinuclear antibodies and anti-NCS/chromatin antibodies. RESULTS: After a mean follow-up period of 45.7 months, anti-NCS/chromatin antibodies were detected in only one patient (2.8%, who developed features of SLE including polyarthritis, lymphopenia, optic neuritis, multiple sclerosis-like lesions, and an autoantibody profile suggestive of SLE. CONCLUSION: The frequency of anti-NCS/chromatin antibodies in primary APS patients is very low, and they may be associated with the development of SLE manifestations.

  8. Serological Testing in Screening for Adult Celiac Disease

    Directory of Open Access Journals (Sweden)

    Helen Rachel Gillett


    Full Text Available Assays for celiac-related antibodies are becoming widely available, and the present review aims to clarify the use of these investigations in the diagnosis of, management of and screening for adult celiac disease. The sensitivities and specificities of various antibody tests are discussed, along with their clinical use as an adjunct to small bowel biopsy, and as a first-line investigation for patients with atypical symptoms of celiac disease or patients at high risk of developing sprue.

  9. Modification and identification of a vector for making a large phage antibody library

    Institute of Scientific and Technical Information of China (English)

    ZHANG Guo-min; CHEN Yü-ping; GUAN Yuan-zhi; WANG Yan; AN Yun-qing


    Background The large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies.Methods scFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC Ⅲ encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated.Results The phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression.Conclusions The reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy.

  10. Antibody performance in western blot applications is context-dependent. (United States)

    Algenäs, Cajsa; Agaton, Charlotta; Fagerberg, Linn; Asplund, Anna; Björling, Lisa; Björling, Erik; Kampf, Caroline; Lundberg, Emma; Nilsson, Peter; Persson, Anja; Wester, Kenneth; Pontén, Fredrik; Wernérus, Henrik; Uhlén, Mathias; Ottosson Takanen, Jenny; Hober, Sophia


    An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13 000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data.

  11. Antiphospholipid antibodies in Brazilian hepatitis C virus carriers

    Directory of Open Access Journals (Sweden)

    A.M. Atta


    Full Text Available Hepatitis C, a worldwide viral infection, is an important health problem in Brazil. The virus causes chronic infection, provoking B lymphocyte dysfunction, as represented by cryoglobulinemia, non-organ-specific autoantibody production, and non-Hodgkin's lymphoma. The aim of this research was to screen for the presence of antiphospholipid autoantibodies in 109 Brazilian hepatitis C virus carriers without clinical history of antiphospholipid syndrome. Forty healthy individuals were used as the control group. IgA, IgG, and IgM antibodies against cardiolipin and β2-glycoprotein I were measured with an enzyme-linked immunosorbent assay, using a cut-off point of either 20 UPL or 20 SBU. While 24 (22.0% hepatitis C carriers had moderate titers of IgM anticardiolipin antibodies (median, 22.5 MPL; 95%CI: 21.5-25.4 MPL, only three carriers (<3% had IgG anticardiolipin antibodies (median, 23 GPL; 95%CI: 20.5-25.5 GPL. Furthermore, IgA anticardiolipin antibodies were not detected in these individuals. Male gender and IgM anticardiolipin seropositivity were associated in the hepatitis C group (P = 0.0004. IgA anti-β2-glycoprotein-I antibodies were detected in 29 of 109 (27.0% hepatitis C carriers (median, 41 SAU; 95%CI: 52.7-103.9 SAU. Twenty patients (18.0% had IgM anti-β2-glycoprotein I antibodies (median, 27.6 SMU; 95%CI: 23.3-70.3 SMU, while two patients had IgG antibodies against this protein (titers, 33 and 78 SGU. Antiphospholipid antibodies were detected in only one healthy individual, who was seropositive for IgM anticardiolipin. We concluded that Brazilian individuals chronically infected with hepatitis C virus present a significant production of antiphospholipid antibodies, mainly IgA anti-β2-glycoprotein I antibodies, which are not associated with clinical manifestations of antiphospholipid syndrome.

  12. Screening and identification of dengue virus-specific antigens and the establishment of ELISA detection method for dengue antibody%登革病毒特异性抗原片段的筛选鉴定及其ELISA方法的建立

    Institute of Scientific and Technical Information of China (English)

    唐博恒; 任瑞文; 洪文艳; 方美玉


    目的 筛选、鉴定登革病毒共同及型特异性抗原,用纯化抗原建立检测登革病毒抗体的ELISA方法.方法 分别利用DNAStar及ANTHEPROT软件对登革病毒1~4型、流行性乙型脑炎及黄热病毒M、E、NS1蛋白进行分析,预测可能的抗原表位.并根据表位位置和氨基酸序列的相似性,分析登革病毒的共有及型特异性抗原表位,并参考GenBank中的序列信息进行比对,分析其在不同登革病毒株中的保守性.然后选择预测得分值较高的表位,利用pET32a、pMAl-c2x原核表达系统进行原核表达,Western blot验证其免疫学反应特异性.Western blot检测阳性抗原片段经亲和纯化后,包被ELISA微孔板,并对ELISA反应条件进行系统优化.结果经系统的生物信息学分析,共预测获得登革病毒抗原表位18个,登革病毒型特异性抗原表位25个,并对其中得分值较高的5段进行了高效表达,经Western blot分析,获得登革1~4型(Den-Ag5),登革2、4型(Den-Ag3),登革1~3型(Den-Ag2)病毒共同抗原片段各一段,登革1、2、4型( Den-Ag1、Den-Ag4)共同抗原片段两段,与流行性乙型脑炎病毒、黄热病毒及所用甲病毒多克隆抗体均无交叉反应.选择Den-Ag5、Den-Ag1和Den-Ag2作为检测用抗原,建立了检测登革病毒抗体的ELISA方法,初步应用表明,所建立方法具备良好特异性,可检测50~200倍稀释的患者血清,S/N比值均在15以上.结论 经系统筛选,获登革病毒特异性抗原片段5段,并建立了检测登革病毒抗体的ELISA方法.%Objective To screen and identify the dengue virus-specific antigens,then establish the ELISA detection method for dengue virus antibody.Methods Using bioinformatic software DNAStar and ANTHEPROT to analyze the hydrophilicity,flexibility,surface probability and antigenicity of dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus M,E and NSI protein amino acid sequence and also consider the influence of

  13. Antibody affinity maturation

    DEFF Research Database (Denmark)

    Skjødt, Mette Louise

    surface expression of various antibody formats in the generated knockout strain. Functional scFv and scFab fragments were efficiently displayed on yeast whereas impaired chain assembly and heavy chain degradation was observed for display of full-length IgG molecules. To identify the optimal polypeptide...... linker for yeast surface display of scFv and scFab fragments, we compared a series of different Gly-Ser-based linkers in display and antigen binding proficiency. We show that these formats of the model antibody can accommodate linkers of different lengths and that introduction of alanine or glutamate...... fragments by in vivo homologous recombination large combinatorial antibody libraries can easily be generated. We have optimized ordered assembly of three CDR fragments into a gapped vector and observed increased transformation efficiency in a yeast strain carrying a deletion of the SGS1 helicase...

  14. Antibody informatics for drug discovery

    DEFF Research Database (Denmark)

    Shirai, Hiroki; Prades, Catherine; Vita, Randi;


    to the antibody science in every project in antibody drug discovery. Recent experimental technologies allow for the rapid generation of large-scale data on antibody sequences, affinity, potency, structures, and biological functions; this should accelerate drug discovery research. Therefore, a robust bioinformatic...... infrastructure for these large data sets has become necessary. In this article, we first identify and discuss the typical obstacles faced during the antibody drug discovery process. We then summarize the current status of three sub-fields of antibody informatics as follows: (i) recent progress in technologies...... for antibody rational design using computational approaches to affinity and stability improvement, as well as ab-initio and homology-based antibody modeling; (ii) resources for antibody sequences, structures, and immune epitopes and open drug discovery resources for development of antibody drugs; and (iii...

  15. Systemic Lupus Erythematosus and Antineutrophil Cytoplasmic Antibody-Associated Vasculitis Overlap Syndrome in Patients With Biopsy-Proven Glomerulonephritis. (United States)

    Jarrot, Pierre-Andre; Chiche, Laurent; Hervier, Baptiste; Daniel, Laurent; Vuiblet, Vincent; Bardin, Nathalie; Bertin, Daniel; Terrier, Benjamin; Amoura, Zahir; Andrés, Emmanuel; Rondeau, Eric; Hamidou, Mohamed; Pennaforte, Jean-Loup; Halfon, Philippe; Daugas, Eric; Dussol, Bertrand; Puéchal, Xavier; Kaplanski, Gilles; Jourde-Chiche, Noemie


    The aim of the study was to report the clinical, biological, and pathological characteristics of patients with glomerulonephritis (GN) secondary to systemic lupus erythematosus (SLE)/antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) overlap syndrome.A nationwide survey was conducted to identify cases of SLE/AAV overlap syndrome. Data were collected from SLE and AAV French research groups. Inclusion criteria were diagnosis of both SLE and AAV according to international classification criteria and biopsy-proven GN between 1995 and 2014. Additional cases were identified through a systematic literature review. A cohort of consecutive biopsy-proven GN was used to study the prevalence of overlapping antibodies and/or overlap syndrome.The national survey identified 8 cases of SLE/AAV overlap syndrome. All patients were female; median age was 40 years. AAV occurred before SLE (n = 3), after (n = 3), or concomitantly (n = 2). Six patients had rapidly progressive GN and 3/8 had alveolar hemorrhage. All patients had antinuclear antibodies (ANA); 7/8 had p-ANCA antimyeloperoxidase (MPO) antibodies. Renal biopsies showed lupus nephritis (LN) or pauci-immune GN. Remission was obtained in 4/8 patients. A literature review identified 31 additional cases with a similarly severe presentation. In the GN cohort, ANCA positivity was found in 30% of LN, ANA positivity in 52% of pauci-immune GN, with no correlation with pathological findings. The estimated prevalence for SLE/AAV overlap syndrome was 2/101 (2%).In patients with GN, SLE/AAV overlap syndrome may occur but with a low prevalence. Most patients have an aggressive renal presentation, with usually both ANA and anti-MPO antibodies. Further studies are needed to assess shared pathogenesis and therapeutic options.

  16. Antithyroglobulin Antibodies and Antimicrosomal Antibodies in Various Thyroid Diseases

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Gwon Jun; Hong, Key Sak; Choi, Kang Won; Lee, Kyu; Koh, Chang Soon; Lee, Mun Ho; Park, Sung Hoe; Chi, Je Geun; Lee, Sang Kook [Seoul National University College of Medicine, Seoul (Korea, Republic of)


    The authors investigated the incidence of antithyroglobulin antibodies and antibodies and antimicrosomal antibodies measured by tanned red cell hemagglutination method in subjects suffering from various thyroid disorders. 1) In 15 normal patients, neither suffering from any thyroid diseases nor from any other autoimmune disorders, the antithyroglobulin antibodies were all negative, but the antimicrosomal antibody was positive only in one patient (6.7%). 2) The antithyroglobulin antibodies were positive in 31.5% (34 patients) of 108 patients with various thyroid diseases, and the antimicrosomal antibodies were positive in 37.0% (40 patients). 3) of the 25 patients with Graves' diseases, 7 patients (28.0%) showed positive for the antithyroglobulin antibodies, and 9 (36.0%) for the antimicrosomal antibodies. There was no definite differences in clinical and thyroid functions between the groups with positive and negative results. 4) Both antibodies were positive in 16 (88.9%) and 17 (94.4%) patients respectively among 18 patients with Hashimoto's thyroiditis, all of them were diagnosed histologically. 5) Three out of 33 patients with thyroid adenoma showed positive antibodies, and 3 of 16 patients with thyroid carcinoma revealed positive antibodies. 6) TRCH antibodies demonstrated negative results in 2 patients with subacute thyroiditis, but positive in one patient with idiopathic primary myxedema. 7) The number of patients with high titers(>l:802) was 16 for antithyroglobulin antibody, and 62.5% (10 patients) of which was Hashimoto's thyroiditis. Thirteen (65.0) of 20 patients with high titers (>l:802) for antimicrosomal antibody was Hashimoto's thyroiditis. TRCH test is a simple, sensitive method, and has high reliability and reproducibility. The incidences and titers of antithyroglobulin antibody and antimicrosomal antibody are especially high in Hashimoto's thyroiditis.

  17. Prediction of Antibody Epitopes

    DEFF Research Database (Denmark)

    Nielsen, Morten; Marcatili, Paolo


    Antibodies recognize their cognate antigens in a precise and effective way. In order to do so, they target regions of the antigenic molecules that have specific features such as large exposed areas, presence of charged or polar atoms, specific secondary structure elements, and lack of similarity...... to self-proteins. Given the sequence or the structure of a protein of interest, several methods exploit such features to predict the residues that are more likely to be recognized by an immunoglobulin.Here, we present two methods (BepiPred and DiscoTope) to predict linear and discontinuous antibody...

  18. Characterization of Two Human Monoclonal Antibodies Neutralizing Influenza A H7N9 Viruses (United States)

    Wang, Jianmin; Chen, Zhe; Bao, Linlin; Zhang, Weijia; Xue, Ying; Pang, XingHuo; Zhang, Xi


    H7N9 was a cause of significant global health concern due to its severe infection and approximately 35% mortality in humans. By screening a Fab antibody phage library derived from patients who recovered from H7N9 infections, we characterized two human monoclonal antibodies (HuMAbs), HNIgGD5 and HNIgGH8. The epitope of these two antibodies was dependent on two residues in the receptor binding site at positions V186 and L226 of the hemagglutinin glycoprotein. Both antibodies possessed high neutralizing activity. PMID:26063436

  19. Measurement of anti-DFS70 antibodies in patients with ANA-associated autoimmune rheumatic diseases suspicion is cost-effective. (United States)

    Gundín, Simón; Irure-Ventura, Juan; Asensio, Esther; Ramos, David; Mahler, Michael; Martínez-Taboada, Victor; López-Hoyos, Marcos


    The presence of antinuclear antibodies (ANA) is associated with a wide range of ANA-associated autoimmune rheumatic diseases (AARD). The most commonly method used for the detection of ANA is indirect immunofluorescence (IIF) on HEp-2 cells. This method is very sensitive but unspecific. As a consequence, ANA testing on HEp-2 substrates outside a proper clinical specialist framework may lead to inappropriate referrals to tertiary care specialists and, worst case inappropriate and potentially toxic therapy for the patient. Among ANA, isolated anti-DFS70 antibodies represent a potentially important biomarker that can be clinically used to discriminate AARD from non-AARD patients in ANA IIF positive individuals. Therefore, their presence may avoid unnecessary follow-up testing and referrals. In our study, we investigated if the implementation of a new ANA workup algorithm allowing for the identification of anti-DFS70 antibodies is cost-effective through the reduction of both unnecessary follow-up testing and outpatient clinic visits generated by the clinical suspicion of a potential AARD. None of the 181 patients included with a positive monospecific anti-DFS70 antibody result developed SARD during the follow-up period of 10 years. The reduction in number of tests after ANA and anti-DFS70 positive results was significant for anti-ENA (230 vs. 114 tests; p < 0.001) and anti-dsDNA antibodies (448 vs. 114 tests; p < 0.001). In addition, the outpatient clinic visits decreased by 70 % (p < 0.001). In total, the adoption of the new algorithm including anti-DFS70 antibody testing resulted in a cost saving of 60869.53 € for this pilot study. In conclusion, the use of anti-DFS70 antibodies was clearly cost-efficient in our setting.

  20. Compositions, antibodies, asthma diagnosis methods, and methods for preparing antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Hongjun; Zangar, Richard C.


    Methods for preparing an antibody are provided with the method including incorporating 3-bromo-4-hydroxy-benzoic acid into a protein to form an antigen, immunizing a mammalian host with the antigen, and recovering an antibody having an affinity for the antigen from the host. Antibodies having a binding affinity for a monohalotyrosine are provided as well as composition comprising an antibody bound with monohalotyrosine. Compositions comprising a protein having a 3-bromo-4-hydroxy-benzoic acid moiety are also provided. Methods for evaluating the severity of asthma are provide with the methods including analyzing sputum of a patient using an antibody having a binding affinity for monohalotyrosine, and measuring the amount of antibody bound to protein. Methods for determining eosinophil activity in bodily fluid are also provided with the methods including exposing bodily fluid to an antibody having a binding affinity for monohalotyrosine, and measuring the amount of bound antibody to determine the eosinophil activity.

  1. Human germline antibody gene segments encode polyspecific antibodies. (United States)

    Willis, Jordan R; Briney, Bryan S; DeLuca, Samuel L; Crowe, James E; Meiler, Jens


    Structural flexibility in germline gene-encoded antibodies allows promiscuous binding to diverse antigens. The binding affinity and specificity for a particular epitope typically increase as antibody genes acquire somatic mutations in antigen-stimulated B cells. In this work, we investigated whether germline gene-encoded antibodies are optimal for polyspecificity by determining the basis for recognition of diverse antigens by antibodies encoded by three VH gene segments. Panels of somatically mutated antibodies encoded by a common VH gene, but each binding to a different antigen, were computationally redesigned to predict antibodies that could engage multiple antigens at once. The Rosetta multi-state design process predicted antibody sequences for the entire heavy chain variable region, including framework, CDR1, and CDR2 mutations. The predicted sequences matched the germline gene sequences to a remarkable degree, revealing by computational design the residues that are predicted to enable polyspecificity, i.e., binding of many unrelated antigens with a common sequence. The process thereby reverses antibody maturation in silico. In contrast, when designing antibodies to bind a single antigen, a sequence similar to that of the mature antibody sequence was returned, mimicking natural antibody maturation in silico. We demonstrated that the Rosetta computational design algorithm captures important aspects of antibody/antigen recognition. While the hypervariable region CDR3 often mediates much of the specificity of mature antibodies, we identified key positions in the VH gene encoding CDR1, CDR2, and the immunoglobulin framework that are critical contributors for polyspecificity in germline antibodies. Computational design of antibodies capable of binding multiple antigens may allow the rational design of antibodies that retain polyspecificity for diverse epitope binding.

  2. Prediction of antibody persistency from antibody titres to natalizumab

    DEFF Research Database (Denmark)

    Jensen, Poul Erik H; Koch-Henriksen, Nils; Sellebjerg, Finn Thorup;


    In a subgroup of patients with multiple sclerosis natalizumab therapy causes generation of anti-natalizumab antibodies that may be transient or persistent. It is recommended to discontinue natalizumab therapy in persistently antibody-positive patients.......In a subgroup of patients with multiple sclerosis natalizumab therapy causes generation of anti-natalizumab antibodies that may be transient or persistent. It is recommended to discontinue natalizumab therapy in persistently antibody-positive patients....

  3. Monoclonal antibodies in myeloma

    DEFF Research Database (Denmark)

    Sondergeld, P.; van de Donk, N. W. C. J.; Richardson, P. G.;


    The development of monoclonal antibodies (mAbs) for the treatment of disease goes back to the vision of Paul Ehrlich in the late 19th century; however, the first successful treatment with a mAb was not until 1982, in a lymphoma patient. In multiple myeloma, mAbs are a very recent and exciting add...

  4. Antibody Blood Tests (United States)

    ... What do I do if I have a negative blood test (or panel) but I’m still having symptoms? While it is rare, it is possible for patients to have a negative antibody test results and still have celiac disease. ...

  5. Prenatal screening and genetics

    NARCIS (Netherlands)

    Alderson, P.; Aro, A.R.; Dragonas, T.; Ettorre, E.; Hemminki, E.; Jalinoja, P.; Santalahti, P.; Tijmstra, T.


    Although the term 'genetic screening' has been used for decades, this paper discusses how, in its most precise meaning, genetic screening has not yet been widely introduced. 'Prenatal screening' is often confused with 'genetic screening'. As we show, these terms have different meanings, and we exami

  6. Stomach (Gastric) Cancer Screening (United States)

    ... Gastric Cancer Treatment Stomach Cancer Prevention Stomach Cancer Screening Research Stomach (Gastric) Cancer Screening (PDQ®)–Patient Version What is ... from the . There is no standard or routine screening test for stomach cancer. Several types of screening tests have been ...

  7. Prenatal screening and genetics

    DEFF Research Database (Denmark)

    Alderson, P; Aro, A R; Dragonas, T


    Although the term 'genetic screening' has been used for decades, this paper discusses how, in its most precise meaning, genetic screening has not yet been widely introduced. 'Prenatal screening' is often confused with 'genetic screening'. As we show, these terms have different meanings, and we ex...

  8. Significance of anti-HLA and donor-specific antibodies in long-term renal graft survival. (United States)

    Saidman, S


    Numerous studies have demonstrated an association of posttransplant HLA antibodies with decreased long-term graft survival. The presence of C4d deposition in these cases supports the hypothesis that antibody and complement deposition are involved in the pathogenesis of graft failure. Development of HLA antibodies may predate the clinical manifestation of chronic rejection (CR). However, frequency of donor-specific antibody is low when all patients are screened regardless of their graft function, and it may be more valuable to look for antibody only in patients with mild dysfunction. Effective treatment for CR has not been identified, although increased immunosuppression has been shown to decrease antibody levels and stabilize graft function. Many patients have been identified with good graft function despite the presence of circulating donor-specific HLA antibody. Additional studies focusing on the mechanism behind the apparent protection from the detrimental effects of antibody in such patients are needed.

  9. What Is Antiphospholipid Antibody Syndrome? (United States)

    ... page from the NHLBI on Twitter. What Is Antiphospholipid Antibody Syndrome? Antiphospholipid (AN-te-fos-fo-LIP-id) antibody ... weeks or months. This condition is called catastrophic antiphospholipid syndrome (CAPS). People who have APS also are at ...

  10. Red Blood Cell Antibody Identification (United States)

    ... ID, RBC; RBC Ab ID Formal name: Red Blood Cell Antibody Identification Related tests: Direct Antiglobulin Test ; RBC ... I should know? How is it used? Red blood cell (RBC) antibody identification is used as a follow- ...

  11. Lupus anticoagulants and antiphospholipid antibodies (United States)

    ... this page: // Lupus anticoagulants and antiphospholipid antibodies To use the sharing features on this page, please enable JavaScript. Lupus anticoagulants are antibodies against substances in the lining ...

  12. Anti-smooth muscle antibody (United States)

    ... gov/ency/article/003531.htm Anti-smooth muscle antibody To use the sharing features on this page, please enable JavaScript. Anti-smooth muscle antibody is a blood test that detects the presence ...


    Directory of Open Access Journals (Sweden)

    I. V. Goraichuk


    Full Text Available Bovine viral diarrhea is a widespread infection of cattle that has a wide range of clinical symptoms in domestic and wild ruminants. It is a major problem in cattle and causes significant economic losses in the cattle industry. The virus infects bovines of all ages and causes both immunosuppression and reproductive, respiratory and digestive disorders. Persistently infected cattle are the main factor in transmission of the disease between and among herds. Comparative results of antibodies presence received by two methods of enzymoimmunoassay and virus neutralization test are given in the paper. During the work, 1010 samples of blood serum of cattle from three farms in the Kharkiv region were selected and analyzed. Bovine viral diarrhea virus concerning antibodies were found by enzymoimmunoassay in 704 samples (69.7% using commercial kit and in 690 samples (68.3% using in house method. After results clarification by virus neutralization test, bovine viral diarrhea antibodies were found in 712 samples (70.5%. Immunoenzyme analysis is recommended for mass screening of cattle for viral diarrhea occurrence. The results confirm that the sensitivity immunoenzyme analysis satisfies the requirements of the diagnostic methods. Using the neutralization reaction of viruses as the «gold standard» of serological methods, it is appropriate to clarify the results of immunoenzyme analysis. Since the results contain a signi ficant number of false positive results, it is necessary to carry out comprehensive studies using both serological and molecular genetics methods.

  14. Screening for abdominalt aortaaneurisme

    DEFF Research Database (Denmark)

    Lindholt, J S; Juul, Svend; Henneberg, E W;


    rupture. Ultrasonographic screening for AAA takes 10 minutes per scan, and the sensitivity and specificity are high. Ultrasonographic screening for AAA is a reliable, safe and inexpensive method for screening, and screening for AAA is discussed worldwide. One point four percent of deaths among men from 65...... on uncertain assumptions concerning prevalence, incidence and risk of rupture. Therefore a randomized trial screening of 65-73 year old males is taking place in the County of Viborg in Denmark. Udgivelsesdato: 1997-Mar-24...

  15. Screening for abdominalt aortaaneurisme

    DEFF Research Database (Denmark)

    Lindholt, Jes Sanddal; Juul, Søren; Henneberg, E W;


    rupture. Ultrasonographic screening for AAA takes 10 minutes per scan, and the sensitivity and specificity are high. Ultrasonographic screening for AAA is a reliable, safe and inexpensive method for screening, and screening for AAA is discussed worldwide. One point four percent of deaths among men from 65...... on uncertain assumptions concerning prevalence, incidence and risk of rupture. Therefore a randomized trial screening of 65-73 year old males is taking place in the County of Viborg in Denmark....

  16. Type and screen policy in the blood bank: Is AHG cross-match still required? A study at a multispecialty corporate hospital in India

    Directory of Open Access Journals (Sweden)

    Pathak Sangeeta


    Full Text Available Background: Antibodies against only about 25-28 blood group antigens are known to cause hemolytic reactions (HTRs, and red cell antibody screening should detect such clinically significant antibodies. An extension of the antibody screening test is the ′type and screen′ done to detect clinically significant antibodies, omiting the anti-human globulin (AHG cross-match. Aim: The aim of this study was to find out if the type and screen procedure is a safe method for pre-transfusion testing when compared to the AHG cross-match currently in use in India. Materials and Methods: We evaluated data from 45373 patients for whom a total of 61668 units of packed red blood cells (PRBC were cross-matched in the AHG phase using DiaMed; ID cards. An antibody screen was carried out in all the patients using the DiaMed; ID-DiaCell I+II+III. The AHG cross-match was also carried out for all recipients, irrespective of the result of the antibody screen. The results were compared to see if there were any cases where the antibody screening was negative but the AHG cross-match showed incompatibility. Results: Not a single case was found where the antibody screen was negative and AHG cross-match showed incompatibility. In 68 cases the antibody screening was positive. Out of the 68 cases, AHG cross-match was incompatible with at least one unit of PRBC in 41 cases. Conclusion: The screening cell panel adequately detected the clinically significant antibodies in the Indian population in our study. The type and screen policy can be safe, efficient, cost-effective, and beneficial to the transfusion service in India.

  17. Challenges in Antibody Development against Tn and Sialyl-Tn Antigens (United States)

    Loureiro, Liliana R.; Carrascal, Mylène A.; Barbas, Ana; Ramalho, José S.; Novo, Carlos; Delannoy, Philippe; Videira, Paula A.


    The carbohydrate antigens Tn and sialyl-Tn (STn) are expressed in most carcinomas and usually absent in healthy tissues. These antigens have been correlated with cancer progression and poor prognosis, and associated with immunosuppressive microenvironment. Presently they are used in clinical trials as therapeutic vaccination, but with limited success due to their low immunogenicity. Alternatively, anti-Tn and/or STn antibodies may be used to harness the immune system against tumor cells. Whilst the development of antibodies against these antigens had a boost two decades ago for diagnostic use, so far no such antibody entered into clinical trials. Possible limitations are the low specificity and efficiency of existing antibodies and that novel antibodies are still necessary. The vast array of methodologies available today will allow rapid antibody development and novel formats. Following the advent of hybridoma technology, the immortalization of human B cells became a methodology to obtain human monoclonal antibodies with better specificity. Advances in molecular biology including phage display technology for high throughput screening, transgenic mice and more recently molecularly engineered antibodies enhanced the field of antibody production. The development of novel antibodies against Tn and STn taking advantage of innovative technologies and engineering techniques may result in innovative therapeutic antibodies for cancer treatment. PMID:26270678

  18. Antibody Engineering and Therapeutics Conference


    Larrick, James W; Parren, Paul WHI; Huston, James S; Plückthun, Andreas; Bradbury, Andrew; Tomlinson, Ian M; Chester, Kerry A.; Burton, Dennis R.; Adams, Gregory P.; Weiner, Louis M.; Scott, Jamie K.; Alfenito, Mark R; Veldman, Trudi; Reichert, Janice M.


    The Antibody Engineering and Therapeutics conference, which serves as the annual meeting of The Antibody Society, will be held in Huntington Beach, CA from Sunday December 8 through Thursday December 12, 2013. The scientific program will cover the full spectrum of challenges in antibody research and development, and provide updates on recent progress in areas from basic science through approval of antibody therapeutics. Keynote presentations will be given by Leroy Hood (Institute of System Bi...

  19. Structural Characterization of Peptide Antibodies

    DEFF Research Database (Denmark)

    Chailyan, Anna; Marcatili, Paolo


    The role of proteins as very effective immunogens for the generation of antibodies is indisputable. Nevertheless, cases in which protein usage for antibody production is not feasible or convenient compelled the creation of a powerful alternative consisting of synthetic peptides. Synthetic peptides...... can be modified to obtain desired properties or conformation, tagged for purification, isotopically labeled for protein quantitation or conjugated to immunogens for antibody production. The antibodies that bind to these peptides represent an invaluable tool for biological research and discovery...

  20. The IgG antibody reactivity of sera from patients with active chronic hepatitis to a crude liver antigen and liver specific protein (LSP): analysis by ELISA and immunoblotting. (United States)

    Sundin, U; Heigl, Z; Sundqvist, K G


    The antibody reactivity to liver specific protein (LSP) and a crude liver antigen of sera from patients with chronic active hepatitis (CAH) were studied along with other related diseases and healthy individuals. CAH sera containing liver reacting antibodies were selected using an ELISA with a crude liver preparation as antigen and subsequently the specificity was analysed by immunoblotting of SDS-PAGE-separated LSP. The incidence of IgG antibodies to the crude liver antigen and LSP in sera from 15 patients with CAH was 94% and 55% respectively. In the healthy control group (n = 30) the corresponding figures were 3% and 17%. Sera from patients with other autoimmune conditions with considerable reactivity in the crude liver ELISA test were those with antibodies against extractable nuclear antigens (ENA) and thyroid gland antigens, while the anti-nuclear antibody (ANA) group as a whole did not differ from the control group. In immunoblotting of SDS-PAGE-separated crude liver and LSP antigens, the IgG binding pattern of ELISA IgG positive CAH sera and sera from patients with thyroid disease was distinct, with bands corresponding to antigens of molecular weights of 38, 45 and 50 kD which were not observed in ELISA negative CAH sera or in sera from patients with other diseases and among healthy controls.

  1. Synthetic peptides for antibody production

    NARCIS (Netherlands)

    Zegers, N.D.


    Synthetic peptides are useful tools for the generation of antibodies. The use of antibodies as specific reagents in inununochemical assays is widely applied. In this chapter, the application of synthetic peptides for the generation of antibodies is described. The different steps that lead to the uni

  2. Human smooth muscle autoantibody. Its identification as antiactin antibody and a study of its binding to "nonmuscular" cells. (United States)

    Gabbiani, G; Ryan, G B; Lamelin, J P; Vassalli, P; Majno, G; Bouvier, C A; Cruchaud, A; Lüscher, E F


    When human serum containing smooth muscle autoantibodies (SMA) is incubated with extracts containing thrombosthenin (the contractile material of platelets) or thrombosthenin-A (the actin-like moiety of thrombosthenin), it loses its ability to bind to smooth muscle. Such binding is also diminished when SMA serum is incubated with lysed platelets; this effect is not seen if the SMA serum is incubated with intact platelets. The incubation of other autoantibodies (such as antimitochondrial or antinuclear antibodies) with thrombosthenin does not affect their binding to the specific antigens. It appears that SMA is directed against the actin fraction of thrombosthenin-ie, SMA is an antiactin antibody. Hence the name of antiactin autoantibody (AAA) seems more appropriate than smooth muscle autoantibody (SMA). A study of the distribution of antiactin autoantibody binding in rat, rabbit and man shows that several "nonmuscular" structures contain actin under normal conditions; these include megakaryocytes and platelets, normal rat hepatocytes, the brush borders of renal tubules, the periphery of epithelial cells of the intestine, polymorphs and lymphocytes in lymph nodes (but not thymic cortical lymphocytes). In addition, certain cell types (such as granulation tissue fibroblasts, cultivated fibroblasts, hepatocytes or regenerating liver and epidermal cells growing over a skin wound) can reversibly acquire a massive network of actin-containing microfilaments resembling those in smooth muscle.

  3. Elevated levels of serum antibodies against alpha-1, 6-glucan in patients with systemic lupus erythematosus or rheumatoid arthritis

    Institute of Scientific and Technical Information of China (English)

    Hui Dai; Xiao-Ming Gao


    This study was undertaken to investigate whether levels of anti-alpha-1,6-glucan antibodies in human sera correlate with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE).Serum samples were collected from patients with SLE (n =30),RA (n =30) and healthy adult volunteers.IgG,IgA and IgM levels against alpha-1,6-glucan were measured using enzyme linked immunosorbent assays.Anti-alpha-1,6-glucan IgG prevalence was raised in patients with active SLE (73.3%)and RA (60%) compared with healthy controls (13.3%).Strong correlation between anti-alpha-1,6-glucan-IgG levels and anti-perinuclear factor (r =0.642; p< 0.05) in RA patients or anti-nuclear antibodies (r =0.675; p < 0.05)in SLE patients was observed.No significant differences in anti-alpha-1,6-glucan-lgA or-lgM levels were noted between different groups.We conclude that anti-alpha1,6-glucan-lgG levels were significantly elevated in patients with SLE or RA and positively correlated with disease activity.

  4. A human monoclonal antibody to high-frequency red cell antigen Jra. (United States)

    Miyazaki, T; Kwon, K W; Yamamoto, K; Tone, Y; Ihara, H; Kato, T; Ikeda, H; Sekiguchi, S


    A human-mouse heterohybridoma (HMR0921) secreting human monoclonal IgG3, lambda antibody was produced from peripheral blood lymphocytes of a healthy blood donor with serum antibody to Jra, by EBV transformation and hybridization with mouse myeloma cell line P3X63Ag8.653. The reactivity of HMR0921 antibody was assessed by antiglobulin test with a panel of red cells including 14 different rare blood types. Only Jr(a-) red cells were negative. The strict specificity of this antibody to Jra antigen was further confirmed by absorption test with fluorescence flow cytometry. On screening of 28,744 blood donor samples by HMR0921 antibody, we detected 19 agglutination-negative samples, which were confirmed as Jr(a-) by conventional anti-Jra antisera. Therefore, our HMR0921 antibody is extremely useful for detecting rare Jr(a-) blood.

  5. Characterization of immobilization methods of antiviral antibodies in serum for electrochemical biosensors

    Energy Technology Data Exchange (ETDEWEB)

    Huy, Tran Quang, E-mail: [National Institute of Hygiene and Epidemiology (NIHE), No1 Yersin St., Hanoi (Viet Nam); International Training Institute for Materials Science (ITIMS), Hanoi University of Science and Technology (HUST), No1 Dai Co Viet, Hanoi (Viet Nam); Hanh, Nguyen Thi Hong; Van Chung, Pham; Anh, Dang Duc; Nga, Phan Thi [National Institute of Hygiene and Epidemiology (NIHE), No1 Yersin St., Hanoi (Viet Nam); Tuan, Mai Anh, E-mail: [International Training Institute for Materials Science (ITIMS), Hanoi University of Science and Technology (HUST), No1 Dai Co Viet, Hanoi (Viet Nam)


    In this paper, we describes different methods to immobilize Japanese encephalitis virus (JEV) antibodies in human serum onto the interdigitated surface of a microelectrode sensor for optimizing electrochemical detection: (1) direct covalent binding to the silanized surface, (2) binding to the silanized surface via a cross-linker of glutaraldehyde (GA), (3) binding to glutaraldehyde/silanized surface via goat anti-human IgG polyclonal antibody and (4) binding to glutaraldehyde/silanized surface via protein A (PrA). Field emission scanning electron microscopy, Fourier transform infrared spectrometry, and fluorescence microscopy are used to verify the characteristics of antibodies on the interdigitated surface after the serum antibodies immobilization. The analyzed results indicate that the use of protein A is an effective choice for immobilization and orientation of antibodies in serum for electrochemical biosensors. This study provides an advantageous immobilization method of serum containing antiviral antibodies to develop electrochemical biosensors for preliminary screening of viruses in clinical samples from outbreaks.

  6. High-resolution mapping of linear antibody epitopes using ultrahigh-density peptide microarrays

    DEFF Research Database (Denmark)

    Buus, Søren; Rockberg, Johan; Forsström, Björn


    -resolution mapping of linear antibody epitopes, we have used ultrahigh-density peptide microarrays generating several hundred thousand different peptides per array. Using exhaustive length and substitution analysis, we have successfully examined the specificity of a panel of polyclonal antibodies raised against......Antibodies empower numerous important scientific, clinical, diagnostic, and industrial applications. Ideally, the epitope(s) targeted by an antibody should be identified and characterized, thereby establishing antibody reactivity, highlighting possible cross-reactivities, and perhaps even warning...... against unwanted (e.g. autoimmune) reactivities. Antibodies target proteins as either conformational or linear epitopes. The latter are typically probed with peptides, but the cost of peptide screening programs tends to prohibit comprehensive specificity analysis. To perform high-throughput, high...

  7. Analysis of the results of antinuclear antibody and its profile for 312 patients%312例抗核抗体与抗细胞核细胞浆抗体谱检测结果的分析

    Institute of Scientific and Technical Information of China (English)

    车文英; 谭延国; 韦希明


    目的 回顾分析抗核抗体(ANA)与抗细胞核细胞浆抗体谱(ANA谱)在本实验室的应用状况,以及在自身免疫性疾病诊断上的价值.方法 ANA用间接免疫荧光法检测,ANA谱用欧蒙印迹法检测.结果 312例临床标本中,ANA阳性数为123例,阳性率为39.9%(123/312),核型种类主要为核细颗粒型、均质型和核仁型等,滴度各有不同.312例标本同时做ANA谱检测,阳性数为88例,阳性率为25%(88/312).阳性抗体出现次数较多的是抗SSA、Ro52、SSB、CENPB(着丝点B蛋白)和Histone(组蛋白)等,基本上与荧光法测定ANA核型的结果相符.只是当ANA为低滴度(起始滴度1:100)时,ANA谱为阴性结果的可能性较高.结论 用间接免疫荧光法筛查ANA,再用ANA谱作确认分型,可以更准确地确定ANA靶抗原的类型,ANA谱可检测15种针对细胞核和细胞浆的抗体,同传统的抗ENA抗体六项相比,扩大了检测范围,增强了对ANA的分型能力,具有较高的应用价值.

  8. Correlation study on indirect immunofluorescence method and blot method for detecting antinuclear antibodies%间接免疫荧光法与印迹法检测抗核抗体的相关性研究

    Institute of Scientific and Technical Information of China (English)

    左克; 李生勇


    目的 研究间接免疫荧光法(IIF)检测抗核抗体(ANA)与印迹法(LIA)检测ANA谱(ANAS)的诊断结果,并分析二者的一致性及相关性.方法 选取2013年2月至2014年11月在该院风湿科疑为自身免疫性疾病(AID)的住院及门诊患者为研究对象,采用IIF与LIA分别检测ANA与抗核抗体谱ANAS,并对结果进行统计分析.结果 检出ANA阳性共127例,ANAS阳性84例,最终确诊AID 96例,各种特异性抗体均有检出,优势性检验发现总体阳性率差异有统计学意义.结论 使用IIF检测ANA的阳性率高于LIA,但对于疾病的诊断价值,则LIA检测ANAS优于IIF检测ANA,临床上二者不可互相代替,应联合检测,防止漏检.

  9. 抗核抗体谱的检测在自身免疫性疾病中的应用价值%Application value of antinuclear antibody spectrum detection in diagnosis of autoimmune diseases

    Institute of Scientific and Technical Information of China (English)

    蒋新颖; 金玉; 张颖; 冯潜


    目的 探讨用间接免疫荧光法(IIF)抗核抗体(ANA)与免疫印迹法(LIA)特异性抗核抗体谱(ANAs)的联合检测在自身免疫性疾病(AID)中的应用价值.方法 436例患者,其中自身免疫病患者271例,非自身免疫病患者165例,所有患者同时检测血清ANA和ANAs,两种检测方法产生4种检出模式:IIF(+)/LIA(+)、IIF(-)/LIA(-)、IIF(+)/LIA(-)和IIF(-)/LIA(+).结果 436份标本中IIF(+)/LIA(+)占40.15%,IIF (-)/LIA(-)占25.52%,IIF(+)/LIA(-)占15.27%,IIF(-)/LIA(+)占9.81%,IIF与LIA检测ANA的结果总体符合率为65.67%,ANA和ANAs在自身免疫病患者中的阳性率分别是65.3%和57.9%,显著高于非自身免疫病患者的17.6%和20.0%;ANA和ANAs在自身免疫病组和非自身免疫病组间阳性率比较,差异有统计学意义(P<0.01).两检测结果仅在MCTD和SLE患者中存在相关性(P<0.01),在其他观察组中不存在相关性(P>0.05).结论 IIF检测ANA容易导致AID患者部分具有临床意义的ANA特异性抗体漏检,而ANAs检测因其测定的抗体数量有限也容易导致AID患者的ANA漏检.IIF-ANA和LIA-ANAs检测不能相互代替,对需要通过检测ANA来排除AID的患者标本应同时进行IIF-ANA和LIF-ANAs的检测,以避免仅采用一种方法进行检测时导致AID患者漏诊.ANA的IIF法易导致以抗-SSA、抗-SSB和抗-Ro52为主要抗体的患者ANA假阴性,而LIA法特异性ANAs的检测因检测的抗体不全面也无法取代ANA的IIF法检测.在临床实际工作中两种ANA的检测方法不能相互取代,应联合应用.

  10. Analysis on detection results of different dilution titers of antinuclear antibodies in children%儿童抗核抗体不同稀释度检测结果分析

    Institute of Scientific and Technical Information of China (English)

    庞舒尹; 陈奕豪; 刘海英; 陈曲波; 蔡莉; 彭小芳; 高飞; 代玉梅; 刘云锋


    目的:比较不同血清稀释滴度对儿童抗核抗体间接免疫荧光分析(A N A‐IIF )结果的影响,探讨降低血清起始稀释度必要性。方法以间接免疫荧光法检测110例健康儿童系列稀释度血清标本抗核抗体(ANA ),并与特异性 ANA 线性免疫(ANA‐LIA)结果相比较;同时分析一组ANA‐IIF阴性的临床患儿标本ANA‐LIA结果。结果健康组标本随着稀释度从1∶80、1∶40、1∶20逐步减低,ANA‐IIF阳性检出率有所上升,分别为7.3%、9.1%、10.9%,但差异无统计学意义(P>0.05),弱阳性分别为7.3%、15.5%、31.8%,差异有统计学意义(P<0.01)。110例健康体检儿童中8例标本检测出特异性ANA,阳性率为7.3%。8例IIF法稀释度1∶80阳性者中,特异性ANA阳性者2例;稀释度1∶40、1∶20新增的4例荧光ANA阳性标本中,有1例特异性ANA阳性。若视ANA‐IIF(1∶80)或ANA‐LIA任一阳性为ANA阳性,ANA阳性率从7.3%上升到12.7%。29例ANA‐IIF(1∶80)为阴性的自身免疫肝病相关自身抗体检测患儿临床标本中,特异性ANA‐LIA检测出5例阳性(17.2%)。结论降低儿童血清起始滴度并不能明显提高ANA‐IIF阳性检出率,反而增加了非特异的弱阳性,因此,临床实验室不需改变儿童ANA常规标本稀释滴度,联合特异性ANA‐LIA的检测有利于ANA的发现。%Objective To compare the influences of different dilution titers on the ANA detection by the indirect immunofluores‐cence assay(IIF) in children for investigating the necessity of reducing serum initial dilution titer .Methods Serum ANA was detec‐ted by using the indirect immunofluorescence assay at a serial of dilution titer in 110 healthy controls and the results were compared with the results of specific ANAs by the linear immunoassay (LIA);meanwhile the ANA‐LIA results in clinical children patients with ANA‐IIF negative were also analyzed .Results With the dilution titers gradual decrease from 1∶80 ,1∶40 and 1∶20 in the samples of the health group ,the positive detection rates of ANA‐IIF were risen ,which were 7 .3% ,9 .1% and 10 .9% respectively , but the differences were not statistically significant (P>0 .05) ,the weak‐positive rates were 7 .3% ,15 .5% and 31 .8% respective‐ly ,the differences were statistically significant (P<0 .01) .Among 110 healthy children under going the physical examination ,the specific ANA was detected out in 8 samples ,the positive rate was 7 .3% .Among 8 positive cases at the dilution titer of 1∶80 by the IIF method ,specific ANA was in 2 cases;in 4 added cases of fluorescence ANA positive samples at the dilution titers of 1∶40 and 1∶20 ,specific ANA was in 1 case .If with any positive of ANA‐IIF(1∶80) or ANA‐LIA as the ANA positive ,the ANA positive rate was risen from 7 .3% to 12 .7% .In the clinical samples among 29 cases of ANA‐IIF(1∶80) negative autoimmune liver disease related autoantibody detection ,the specific ANA‐LIA positive was detected in 5 cases (17 .2% ) .Conclusion Reducing the initial ti‐ter of children serum is unable to obviously increase the ANA‐IIF positive detection rate ,on the contrary increases the non‐specific weak positive .Therefore ,clinical laboratory does not change the dilution titer of children routine ANA sample .The detection by combining with the specific ANA‐LIA spectrum is conducive to find ANA .

  11. Identification of anti-HPA-1a allo-antibodies using IgG platelet antibody detection and crossmatch system assay with Galileo Echo. (United States)

    Di Cristofaro, Julie; Frassati, Coralie; Montagnie, Rolande; Basire, Agnes; Merieux, Yves; Picard, Christophe


    Fetal/neonatal allo-immune thrombocytopenia is the most frequent and the most dangerous clinical condition involving anti-human platelet antigens (HPA)-1a allo-antibodies. Anti-HPA-1a allo-immunization requires rapid and accurate diagnosis to determine appropriate treatment. The Capture-P Ready-Screen assay (C-PRS) is a new qualitative immunoassay to detect IgG anti-human leukocyte antigen (HLA) and anti-HPA allo-antibodies. The aim of this study is to assess the identification of anti-HPA-1a allo-antibodies using the C-PRS assay, associated with HLA class I stripping reagents, on the automated benchtop analyzer Galileo Echo. Forty-nine sera were analyzed: without anti-HLA class I or anti-HPA allo-antibodies, with anti-HLA class I allo-antibodies, with anti-HPA-1a allo-antibodies, among which with anti-HLA class I allo-antibodies. None of the samples without allo-antibodies were reactive. Only anti-HLA antibodies, detected by cytotoxicity-dependent complement and not by Luminex, remained positive before and after stripping reagents. Of the 13 samples, anti-HPA-1a allo-antibodies that were correctly identified before and after incubation with HLA assassin reagent were 70% and 85%, respectively. Anti-glycoprotein auto-antibodies and anti-HLA allo-antibodies do not interfere with the detection of anti-HPA-1a antibodies. This preliminary study indicates that further improvement of the test will be helpful in developing a clinically useful assay in the future.

  12. Antiphospholipid Antibody and Antiphospholipid Syndrome

    Institute of Scientific and Technical Information of China (English)



    @@ Antiphospholipid antibodies (APA) APA is a big category for all kinds of negative charge phospholipid or lecithin - a protein complex autoantibodies or the same antibody, through its recognition of antigen (target protein) different, and phospholipids or lecithin - protein complex combination of various rely on the interference Phospholipid clotting and anti-coagulation factor, and promote endothelial cells, platelets, complement activation and play a role. APA including lupus anticoagulant(LA) and anticardiolipin antibody (ACA), In addition, there are anti-β2 glycoprotein-I (β2-GPI) antibody, anti-prothrombin (a- PT) antibody, anti-lysophosphatidic acid antibody and anti-phosphatidylserine antibody, and so on. APA as the main target of phospholipid-binding protein, including β2-GPI, prothrombin, annexin, protein C (PC) and protein S (PS), plasminogen, and so on.

  13. Spectrophotometric Enzyme Assays for High-Throughput Screening

    Directory of Open Access Journals (Sweden)

    Jean-Louis Reymond


    Full Text Available This paper reviews high-throughput screening enzyme assays developed in our laboratory over the last ten years. These enzyme assays were initially developed for the purpose of discovering catalytic antibodies by screening cell culture supernatants, but have proved generally useful for testing enzyme activities. Examples include TLC-based screening using acridone-labeled substrates, fluorogenic assays based on the β-elimination of umbelliferone or nitrophenol, and indirect assays such as the back-titration method with adrenaline and the copper-calcein fluorescence assay for aminoacids.

  14. Video Screen Capture Basics (United States)

    Dunbar, Laura


    This article is an introduction to video screen capture. Basic information of two software programs, QuickTime for Mac and BlueBerry Flashback Express for PC, are also discussed. Practical applications for video screen capture are given.

  15. Cervical Cancer Screening (United States)

    ... Cancer found early may be easier to treat. Cervical cancer screening is usually part of a woman's health ... may do more tests, such as a biopsy. Cervical cancer screening has risks. The results can sometimes be ...

  16. Alcohol Use Screening (United States)

    ... Centers Diseases + Condition Centers Mental Health Medical Library Alcohol Use Screening (AUDIT-C) - Instructions The following questions ... this tool, there is also text-only version . Alcohol Use Screening (AUDIT-C) - Manual Instructions The following ...

  17. Hearing Loss: Screening Newborns (United States)

    ... of this page please turn JavaScript on. Feature: Hearing Loss Screening Newborns Past Issues / Spring 2015 Table of ... of newborns in the U.S. are screened for hearing loss before they leave the hospital. Research improves the ...

  18. Screening for Ovarian Cancer (United States)

    Understanding Task Force Recommendations Screening for Ovarian Cancer The U.S. Preventive Services Task Force (Task Force) has issued a final recommendation on Screening for Ovarian Cancer . This recommendation is for ...

  19. Screening for Glaucoma (United States)

    Understanding Task Force Recommendations Screening for Glaucoma The U.S. Preventive Services Task Force (Task Force) has issued a final recommendation statement on Screening for Glaucoma . This final recommendation statement ...

  20. Efficient method to optimize antibodies using avian leukosis virus display and eukaryotic cells. (United States)

    Yu, Changming; Pike, Gennett M; Rinkoski, Tommy A; Correia, Cristina; Kaufmann, Scott H; Federspiel, Mark J


    Antibody-based therapeutics have now had success in the clinic. The affinity and specificity of the antibody for the target ligand determines the specificity of therapeutic delivery and off-target side effects. The discovery and optimization of high-affinity antibodies to important therapeutic targets could be significantly improved by the availability of a robust, eukaryotic display technology comparable to phage display that would overcome the protein translation limitations of microorganisms. The use of eukaryotic cells would improve the diversity of the displayed antibodies that can be screened and optimized as well as more seamlessly transition into a large-scale mammalian expression system for clinical production. In this study, we demonstrate that the replication and polypeptide display characteristics of a eukaryotic retrovirus, avian leukosis virus (ALV), offers a robust, eukaryotic version of bacteriophage display. The binding affinity of a model single-chain Fv antibody was optimized by using ALV display, improving affinity >2,000-fold, from micromolar to picomolar levels. We believe ALV display provides an extension to antibody display on microorganisms and offers virus and cell display platforms in a eukaryotic expression system. ALV display should enable an improvement in the diversity of properly processed and functional antibody variants that can be screened and affinity-optimized to improve promising antibody candidates.

  1. Production of anti-idiotype antibodies for deoxynivalenol and their evaluation with three immunoassay platforms. (United States)

    Maragos, C M


    Immunoassays for deoxynivalenol (DON) that involve binding to DON-specific antibodies have been widely developed. In such assays, the responses of samples are generally compared with calibration curves generated by using DON in competition with labeled reagents such as enzymatic or fluorescent conjugates of the toxin. However, materials that mimic the toxin can also be used, provided that they compete effectively with the labeled reagents for the DON-specific antibodies. Examples include certain types of anti-idiotype antibodies, obtained by the immunization of animals with toxin-specific antibodies. In the present work, anti-idiotype antibodies were developed which mimicked DON in the ability to bind to a DON-specific monoclonal antibody (Mab). Fab fragments of the Mab (Ab1) were used to immunize rabbits. Sera were screened by competitive direct enzyme linked immunosorbent assay (CD-ELISA) for the presence of anti-idiotype antibodies (Ab2). In order to determine the most effective screening format and also the potential efficacy in various forms of biosensors, the sera were further evaluated in biolayer interferometry (BLI) and fluorescence polarization immunoassay (FPIA) formats. All three formats were used to demonstrate the presence of anti-idiotypes capable of binding to the paratope of the DON antibody (subtypes Ab2β or Ab2γ). Such materials have the potential to replace DON as calibrants in immunoassays for this toxin.


    Directory of Open Access Journals (Sweden)



    Full Text Available A total of 1021 randomly selected serum samples of adult male goats that had been screened for trypanosome infection were assayed for sperm antibodies using the immunoperoxidase staining technique. The result of the trypanosome screening revealed that 586(57.39% goats were positive for trypanosome infection, while 435(42.61% were negative. The assay for sperm antibodies showed that 482(47.21% animals were positive, while 539(52.79% were negative. In the group that was positive for trypanosome infection, 364(62.12% animals were positive, whereas 222(37.88% were negative for sperm antibodies (P<0.001. The group that was negative for trypanosome infection, had a significantly lower number and proportion 118(27.13% of positive compared to 317(72.87% negative for sperm antibodies. Out of a total 482 goats that were positive for sperm antibodies, a significantly higher number, 364(75.52%, were positive than 118(24.48% that were negative for trypanosome infection (P<0.001. In the group that was found negative for sperm antibodies, a significantly lower proportion, 222(41.19%, was positive compared to 317(58.81% that were negative for trypanosome infection (P<0.001. Seropositivity to sperm antibodies was positively correlated to trypanosome infection (P<0.001. Further work on the pathogenesis of sperm antibody production in trypanosome infection is advocated.

  3. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye. (United States)

    Nath, Nidhi; Godat, Becky; Zimprich, Chad; Dwight, Stephen J; Corona, Cesear; McDougall, Mark; Urh, Marjeta


    beneficial for screening a large number of antibody samples during early monoclonal development phase.

  4. Microfluidic assay without blocking for rapid HIV screening and confirmation. (United States)

    Song, Lusheng; Zhang, Yi; Wang, Wenjun; Ma, Liying; Liu, Yong; Hao, Yanlin; Shao, Yiming; Zhang, Wei; Jiang, Xingyu


    The essential step for HIV spreading limitation is the screening tests. However, there are multiple disadvantages in current screening assays which need further confirmation test. Herein we developed a rapid HIV assay combining screening and confirmation test by using the microfluidic network assay. Meanwhile, the assay is accelerated by bypassing the step of blocking. We call this method as microfluidic assay without blocking (MAWB). Both the limit of detection and reagent incubation time of MAWB are determined by screening of one model protein pair: ovalbumin and its antibody. The assay time is accelerated about 25% while the limit of detection (LOD) is well kept. Formatting the method in for both HIV screening (testing 8 HIV-related samples) and confirmation (assaying 6 kinds of HIV antibodies of each sample) within 30 min was successful. Fast HIV screening and confirmation of 20 plasma samples were also demonstrated by this method. MAWB improved the assay speed while keeping the LOD of conventional ELISA. Meanwhile, both the accuracy and throughput of MAWB were well improved, which made it an excellent candidate for a quick HIV test for both screening and confirmation. Methods like this one will find wide applications in clinical diagnosis and biochemical analysis based on the interactions between pairs of molecules.

  5. Colorectal cancer screening. (United States)

    Burt, Randall W; Cannon, Jamie A; David, Donald S; Early, Dayna S; Ford, James M; Giardiello, Francis M; Halverson, Amy L; Hamilton, Stanley R; Hampel, Heather; Ismail, Mohammad K; Jasperson, Kory; Klapman, Jason B; Lazenby, Audrey J; Lynch, Patrick M; Mayer, Robert J; Ness, Reid M; Provenzale, Dawn; Rao, M Sambasiva; Shike, Moshe; Steinbach, Gideon; Terdiman, Jonathan P; Weinberg, David; Dwyer, Mary; Freedman-Cass, Deborah


    Mortality from colorectal cancer can be reduced by early diagnosis and by cancer prevention through polypectomy. These NCCN Guidelines for Colorectal Cancer Screening describe various colorectal screening modalities and recommended screening schedules for patients at average or increased risk of developing colorectal cancer. In addition, the guidelines provide recommendations for the management of patients with high-risk colorectal cancer syndromes, including Lynch syndrome. Screening approaches for Lynch syndrome are also described.

  6. Screen time and children (United States)

    ... obesity ) Screen time increases your child's risk of obesity because: Sitting and watching a screen is time that is not spent being physically active. TV commercials and other screen ads can lead to unhealthy food choices . Most of the time, the foods in ads ...

  7. Breast awareness and screening. (United States)

    Harmer, Victoria

    Breast cancer is the most commonly diagnosed cancer in the UK. Breast awareness and screening, along with better treatment, can significantly improve outcomes, and more women than ever are now surviving the disease. This article discusses breast awareness and screening, symptoms and risk factors for breast cancer, and how nurses can raise breast awareness and screening uptake.

  8. Screen Practice in Curating

    DEFF Research Database (Denmark)

    Toft, Tanya Søndergaard


    During the past one and a half decade, a curatorial orientation towards "screen practice" has expanded the moving image and digital art into the public domain, exploring alternative artistic uses of the screen. The emergence of urban LED screens in the late 1990s provided a new venue that allowed...

  9. Between Stage and Screen

    NARCIS (Netherlands)

    Tornqvist, Egil


    Ingmar Bergman is worldwide known as a film and stage director. Yet no-one has attempted to compare his stage and screen activities. In Between stage and screen Egil Tornqvist examines formal and thematical correspondences and differences between a number of Bergman's stage, screen, and radio produc

  10. Seroepidemiologic studies in Oaxaca, Mexico. II. Survey for arbovirus antibody. (United States)

    Goldsmith, R S; Zarate, M L; Cedeño-Ferreira, J; Antonio Paz, E


    A serologic survey was conducted in south-western Mexico to obtain information on human experience with arbovirus infections. Sera were collected from two semitropical areas along the Pacific coast of Oaxaca State, two mountain areas above 1,700 meters and the interior valley at 1,500 meters. Of the 610 sera tested for group A antibody, 4.9 per cent were positive in the hemagglutination-inhibition (HI) test to Venezuelan (VE), 11 per cent to Eastern, and none to Western encephalitis viruses. In neutralization tests the antibody was shown to be probably due to VE virus infections. When sera were screened for group B antibodies in the HI test, 32 per cent were positive with St. Louis encephalitis (SLE), 19 per cent with Ilhéus, and 4 per cent with yellow fever viruses. The pattern of reactions suggested that SLE or an antigenically related virus was responsible for the antibody detected. An unusually high rate was found in a mountain area at 2,000 meters: 41 per cent of 113 persons tested were seropositive to SLE. Of 493 sera screened by complement-fixation test, 6 per cent were positive to Nepuyo, 4 per cent to Patois, and 3 per cent to Tlacotalpan viruses.

  11. Circulating protein and antibody biomarker for personalized cancer immunotherapy. (United States)

    Yuan, Jianda


    Immune checkpoint blockade therapies are revolutionizing standard cancer treatments. Immune checkpoint inhibitors likely function to enhance the tumor specific antigen response in order to achieve favorable clinical outcomes. Thus, continuous efforts to identify the common tumor-specific antigens are essential for the broad clinical application of these therapies. Several immunoproteomics approaches have been used in order to screen for this specificity. In a recent article from Jhaveri and colleagues published in the February issue of Cancer Immunology Research, antibody biomarkers were screened in pancreatic cancer patients who received allogeneic, granulocyte-macrophage colony stimulating factor-secreting pancreatic cancer vaccine (GVAX) by using a serum antibody-based SILAC immunoprecipitation (SASI) approach. Using this assay, several new tumor antigens (MYPT1, PSMC5 and TRFR) were identified that were found to have significantly different expression in tumors compared with normal tissue. Moreover, patients with detectable antibodies showed improved disease-free survival after GVAX therapy. These targets need to be further validated to determine the full spectrum of tumor antigen immunogencity and their potential clinical application. In addition to antibodies, circulating protein, DNA and RNA in peripheral blood are under clinical investigation as liquid biopsies and have the potential to provide guidance for future personalized cancer immunotherapy.

  12. How antibodies use complement to regulate antibody responses. (United States)

    Sörman, Anna; Zhang, Lu; Ding, Zhoujie; Heyman, Birgitta


    Antibodies, forming immune complexes with their specific antigen, can cause complete suppression or several 100-fold enhancement of the antibody response. Immune complexes containing IgG and IgM may activate complement and in such situations also complement components will be part of the immune complex. Here, we review experimental data on how antibodies via the complement system upregulate specific antibody responses. Current data suggest that murine IgG1, IgG2a, and IgG2b upregulate antibody responses primarily via Fc-receptors and not via complement. In contrast, IgM and IgG3 act via complement and require the presence of complement receptors 1 and 2 (CR1/2) expressed on both B cells and follicular dendritic cells. Complement plays a crucial role for antibody responses not only to antigen complexed to antibodies, but also to antigen administered alone. Lack of C1q, but not of Factor B or MBL, severely impairs antibody responses suggesting involvement of the classical pathway. In spite of this, normal antibody responses are found in mice lacking several activators of the classical pathway (complement activating natural IgM, serum amyloid P component (SAP), specific intracellular adhesion molecule-grabbing non-integrin R1 (SIGN-R1) or C-reactive protein. Possible explanations to these observations will be discussed.

  13. Evaluation of rubella igg antibodies among women at marriage in kermanshah city, before and after mass vaccination

    Directory of Open Access Journals (Sweden)

    Hossein Hatami


    Conclusion: In our previous study, we had recommended to screen for susceptibility to rubella before marriage, which is no longer required since more than 99% of vaccinated girls showed immunity at the time of marriage. However, as sustainability of immunity after rubella vaccination is usually less than immunity due to illness, we recommend screening for rubella protective antibody every few years.

  14. The antibody Hijikata Tatsumi

    Directory of Open Access Journals (Sweden)

    Éden Peretta


    Full Text Available Considered one of the most influential modern dance representatives in Japan, Tatsumi Hijikata’s work was a milestone in the Japanese post-war experimental artistic scene. Heretic son of his time, he staged a fertile mix of artistic and cultural influences, overlapping subversive elements of European arts and philosophy with radical references from pre-modern Japanese culture. In this way he built the foundations of its unstable antibody, its political-artistic project of dissolution of a organism, both physical and social.

  15. Screening for colorectal cancer. (United States)

    He, Jin; Efron, Jonathan E


    March is national colorectal cancer awareness month. It is estimated that as many as 60% of colorectal cancer deaths could be prevented if all men and women aged 50 years or older were screened routinely. In 2000, Katie Couric's televised colonoscopy led to a 20% increase in screening colonoscopies across America, a stunning rise called the "Katie Couric Effect". This event demonstrated how celebrity endorsement affects health behavior. Currently, discussion is ongoing about the optimal strategy for CRC screening, particularly the costs of screening colonoscopy. The current CRC screening guidelines are summarized in Table 2. Debates over the optimum CRC screening test continue in the face of evidence that 22 million Americans aged 50 to 75 years are not screened for CRC by any modality and 25,000 of those lives may have been saved if they had been screened for CRC. It is clear that improving screening rates and reducing disparities in underscreened communities and population subgroups could further reduce colorectal cancer morbidity and mortality. National Institutes of Health consensus identified the following priority areas to enhance the use and quality of colorectal cancer screening: Eliminate financial barriers to colorectal cancer screening and appropriate follow-up of positive results of colorectal cancer screening. Develop systems to ensure the high quality of colorectal cancer screening programs. Conduct studies to determine the comparative effectiveness of the various colorectal cancer screening methods in usual practice settings. Encouraging population adherence to screening tests and allowing patients to select the tests they prefer may do more good (as long as they choose something) than whatever procedure is chosen by the medical profession as the preferred test.

  16. Increased levels of anti-glycan antibodies in patients with cystic fibrosis

    Directory of Open Access Journals (Sweden)

    Hirche TO


    Full Text Available Abstract Background The prevalence of Crohn's disease (CD is increased in patients with cystic fibrosis (CF. Anti-Saccharomyces cerevisiae antibodies (ASCA have been suggested as a screening tool to detect CD in CF. Recently, several new anti-glycan antibodies have been reported in CD. Materials and methods The sera of 119 CF patients of various age groups were prospectively screened for ASCA type IgG (gASCA, anti-laminaribioside carbohydrate IgG antibodies (ALCA, anti-chitobioside carbohydrate IgA antibodies (ACCA, and anti-mannobioside carbohydrate IgG antibodies (AMCA. The frequency of these anti-glycan antibodies was then compared in patients with CD, ulcerative colitis, rheumatoid arthritis and healthy volunteers. Results A significant number of CF patients were positive for gASCA (51.3% [41.6-60.6] and up to three other anti-glycan antibodies concurrently. Serum levels of anti-glycan antibodies in CF and CD were not related to parameters of inflammation. Despite the well-documented difference in clinical course between male and female CF patients no gender difference of anti-glycan antibodies was found. In contrast, there was a significant positive correlation between anti-glycan markers and age in CF patients. Conclusions Our findings demonstrate for the first time the increased frequency of a panel of anti-glycan antibodies in CF and provide a link between the presence of these serological biomarkers and patient's age. Anti-glycan antibody profiling may therefore become a valuable tool in the care of patients with CF.

  17. Cancer imaging with radiolabeled antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Goldenberg, D.M. (Center for Molecular Medicine and Immunology, Newark, NJ (US))


    This book presents a perspective of the use of antibodies to target diagnostic isotopes to tumors. Antibodies with reasonable specificity can be developed against almost any substance. If selective targeting to cancer cells can be achieved, the prospects for a selective therapy are equally intriguing. But the development of cancer detection, or imaging, with radiolabeled antibodies has depended upon advances in a number of different areas, including cancer immunology and immunochemistry for identifying suitable antigen targets and antibodies to these targets, tumor biology for model systems, radiochemistry for he attachment of radionuclides to antibodies, molecular biology for reengineering the antibodies for safer and more effective use in humans, and nuclear medicine for providing the best imaging protocols and instrumentation to detect minute amounts of elevated radioactivity against a background of considerable noise. Accordingly, this book has been organized to address the advances that are being made in many of these areas.


    Directory of Open Access Journals (Sweden)

    S. Saidi


    Full Text Available One hundred sera from children 1 - 6 years of age, representative of a large serum collection, were tested for the prevalence of antibodies against different viruses. Hemagglutination-inhibition (HI antibodies were found in 68% for measles; 61 % for rubella; 75'% for influenza A2/Hong Kong/68, 16% for influenza B/Md./59, 0% for group A arboviruses, 10% for group B arboviruses, 3% for phlebotomus fever group and 4% for Congo-Crimean hemorrhagic fever (C-CHF group of arboviruses Poliomyelitis-neutralizing antibodies for type 1, 2 and 3 were 90%; 85% and 84%~ respectively. Antibody to EH virus was detected in 84% of the sera by immuno-fluorescence. None of the sera were positive for hepatitis-B antigen or antibody by immuno-precipitation test. The prevalence of some viral antibodies found in this survey are compared with results obtained from surveys in other parts of the country.

  19. [Colorectal cancer screening]. (United States)

    Castells, Antoni


    Colorectal cancer is one of malignancies showing the greatest benefit from preventive measures, especially screening or secondary prevention. Several screening strategies are available with demonstrated efficacy and efficiency. The most widely used are the faecal occult blood test in countries with population-based screening programmes, and colonoscopy in those conducting opportunistic screening. The present article reviews the most important presentations on colorectal cancer screening at the annual congress of the American Gastroenterological Association held in Washington in 2015, with special emphasis on the medium-term results of faecal occult blood testing strategies and determining factors and on strategies to reduce the development of interval cancer after colonoscopy.

  20. Characterization of single chain antibody targets through yeast two hybrid

    Directory of Open Access Journals (Sweden)

    Vielemeyer Ole


    Full Text Available Abstract Background Due to their unique ability to bind their targets with high fidelity, antibodies are used widely not only in biomedical research, but also in many clinical applications. Recombinant antibodies, including single chain variable fragments (scFv, are gaining momentum because they allow powerful in vitro selection and manipulation without loss of function. Regardless of the ultimate application or type of antibody used, precise understanding of the interaction between the antibody's binding site and its specific target epitope(s is of great importance. However, such data is frequently difficult to obtain. Results We describe an approach that allows detailed characterization of a given antibody's target(s using the yeast two-hybrid system. Several recombinant scFv were used as bait and screened against highly complex cDNA libraries. Systematic sequencing of all retained clones and statistical analysis allowed efficient ranking of the prey fragments. Multiple alignment of the obtained cDNA fragments provided a selected interacting domain (SID, efficiently narrowing the epitope-containing region. Interactions between antibodies and their respective targets were characterized for several scFv. For AA2 and ROF7, two conformation-specific sensors that exclusively bind the activated forms of the small GTPases Rab6 and Rab1 respectively, only fragments expressing the entire target protein's core region were retained. This strongly suggested interaction with a non-linear epitope. For two other scFv, TA10 and SF9, which recognize the large proteins giantin and non-muscle myosin IIA, respectively, precise antibody-binding regions within the target were defined. Finally, for some antibodies, secondary targets within and across species could be revealed. Conclusions Our method, utilizing the yeast two-hybrid technology and scFv as bait, is a simple yet powerful approach for the detailed characterization of antibody targets. It allows precise

  1. Analysis of Detecting HIV-1 Antibody in Paired Urine and Serum Specimens from Drug Users by ELISA

    Institute of Scientific and Technical Information of China (English)

    刘中夫; 李志军; 刘世亮; 李莉; 梁富雄; 郑锡文


    Objective: To compare the consistency of the results from detecting HIV-1 antibody in the paired urine and serum specimens from drug users by ELISA.Methods: The paired urine and serum specimens from 273 drug users detained at a detoxification unit were collected, and the HIV-1 antibodies in the specimens of them were screened by urine and serum ELISA kits, respectively. Results: Of 273 serum specimens, 94 ones showed positive reaction and among 94 counterpart urine specimens, 93 ones also appeared positive reaction. Taking the results together,the consistent rate of HIV-1 antibody screened by urine and serum ELISA kits was 99.6%.Conclusion: The urine ELISA kit, which screened HIV-1 antibody of urine showing almost the same results tested by serum ELISA kit, is reliable. It is proposed that urine ELISA be introduced in many fields.

  2. A tetra-layer microfluidic system for peptide affinity screening through integrated sample injection. (United States)

    Wang, Weizhi; Huang, Yanyan; Jin, Yulong; Liu, Guoquan; Chen, Yi; Ma, Huimin; Zhao, Rui


    A novel integrated microfluidic system was designed and fabricated for affinity peptide screening with in situ detection. A tetra-layer microfluidic hybrid chip containing two top eccentric diffluent layers, an inter-layer and a bottom screening layer, was developed as the core device of the system. The eccentric diffluent layers were ingeniously invented for the vertical sample delivery from 2 top-inlets into 12 bottom-inlets, which eliminated the use of excessive accessories and complicated pipelines currently used in other systems. By using six pH gradient generators, the magnetic bead-based screening in 36 parallel chambers was simultaneously carried out under 6 different pH conditions from 5.4 to 8.2. This allowed simultaneous screening of 6 compounds and each under 6 different pH conditions. The fabricated chip system was applied to screening of affinity peptides towards β-endorphin antibody. The affinities of the peptide ligands to the antibody were assessed by on-chip confocal detection. The results from the screening study using this system indicated that the pentapeptide with the sequence of YGGFL had the highest affinity towards β-endorphin antibody at pH 7.1, which was further confirmed by the ELISA assay using a 96-well plate format. This microfluidic screening system is automatic, low-cost and reusable for not only peptide screening but also other bioactive compounds screening towards target molecules.

  3. Anti-plasminogen antibodies compromise fibrinolysis and associate with renal histology in ANCA-associated vasculitis. (United States)

    Berden, Annelies E; Nolan, Sarah L; Morris, Hannah L; Bertina, Rogier M; Erasmus, Dianhdra D; Hagen, E Christiaan; Hayes, Donal P; van Tilburg, Nico H; Bruijn, Jan A; Savage, Caroline O S; Bajema, Ingeborg M; Hewins, Peter


    Antibodies recognizing plasminogen, a key component of the fibrinolytic system, associate with venous thrombotic events in PR3-ANCA vasculitis. Here, we investigated the prevalence and function of anti-plasminogen antibodies in independent UK and Dutch cohorts of patients with ANCA-associated vasculitis (AAV). We screened Ig isolated from patients (AAV-IgG) and healthy controls by ELISA. Eighteen of 74 (24%) UK and 10/38 (26%) Dutch patients with AAV had anti-plasminogen antibodies compared with 0/50 and 1/61 (2%) of controls. We detected anti-plasminogen antibodies in both PR3-ANCA- and MPO-ANCA-positive patients. In addition, we identified anti-tissue plasminogen activator (tPA) antibodies in 13/74 (18%) patients, and these antibodies were more common among patients with anti-plasminogen antibodies (P = 0.011). Eighteen of 74 AAV-IgG (but no control IgG) retarded fibrinolysis in vitro, and this associated with anti-plasminogen and/or anti-tPA antibody positivity. Only 4/18 AAV-IgG retarded fibrinolysis without harboring these antibodies; dual-positive samples retarded fibrinolysis to the greatest extent. Patients with anti-plasminogen antibodies had significantly higher percentages of glomeruli with fibrinoid necrosis (P < 0.05) and cellular crescents (P < 0.001) and had more severely reduced renal function than patients without these antibodies. In conclusion, anti-plasminogen and anti-tPA antibodies occur in AAV and associate with functional inhibition of fibrinolysis in vitro. Seropositivity for anti-plasminogen antibodies correlates with hallmark renal histologic lesions and reduced renal function. Conceivably, therapies that enhance fibrinolysis might benefit a subset of AAV patients.

  4. Characterization of Tumor-Avid Antibody Fragments Genetically Engineered for Mono-Specific Radionuclide Chelation

    Energy Technology Data Exchange (ETDEWEB)

    Quinn, T.P.


    The successful clinical application of targeted-radiopharmaceuticals depends on the development of molecules that optimize tumor specific radionuclide deposition and minimize non-specific organ irradiation. To this end, this proposal outlines a research effort to identify and evaluate novel antibodies and antibody fragments that bind breast tumors. The tumor-avid antibodies will be investigated for as imaging and therapeutic agents and to gain a better understanding of the pharmacokinetics and metabolism of radiolabeled tumor-avid antibody fragments through the use of site-specifically labeled molecules. Antibodies or antibody fragments, that bind breast carcinoma carbohydrate antigens, will be obtained from hybridoma or bacteriophage library screening. More specifically, antibody fragments that bind the carcinoma-associated Thomsen-Friedenreich (T) antigen will be radiolabeled with {sup 99m}Tc and {sup 188}Re at a natural amino acid chelation site and will be investigated in vivo for their abilities to target human breast tumors. In addition, site-specific radiolabeled antibody fragments will be biosynthesized using misacylated suppressor tRNAs. Homogeneously radiolabeled populations of antibody fragments will be used to investigate the effects of radionuclide location and chelation chemistries on their biodistribution and metabolism. It is hypothesized that site-specifically radiolabeled antibody fragments will possess enhanced tumor imaging and therapeutic properties due to optimal label location and conjugation chemistries. New insights into the factors that govern antibody metabolism in vivo are also expected from this work. Results from these studies should enhance our ability to design and synthesize radiolabeled antibody fragments that have improved pharmacokinetic properties. The studies in this proposal involve basic research into the development of antibody-based radiopharmaceuticals, with the ultimate goal of application in humans. This type of basic

  5. ANA、抗dsDNA抗体和抗ENA抗体谱对系统性红斑狼疮的诊断价值%The clinical value of ANA,anti-dsDNA antibody and anti-ENA antibody repertoire in systemic lupus erythematosus diagnosis

    Institute of Scientific and Technical Information of China (English)

    谌晓燕; 陈艳; 张银辉; 王霓; 吕春兰


    Objective To investigate clinical value of the anti-nuclear antibody (ANA ) ,anti-double strand DNA (dsDNA ) anti-body and anti-extractable nuclear antigen(ENA) antibody repertoire in systemic lupus erythematosus (SLE) diagnosis .Methods 158 SLE patients were served as SLE group and another 50 healthy people as the control group .Indirect immunofluorescence(IIF) and immunoblotting test(IBT ) were employed to detect ANA ,anti-dsDNA antibody and anti-ENA antibody repertoire .Results Positive rates of ANA ,anti-dsDNA antibody and anti-ENA antibody repertoire in SLE diagnosis were 81 .65% ,68 .35% and 87 .34% ,respectively ,which were all lower than that of their combined detection (93 .04% ) ,with statistically significant difference (P<0 .05 ) .Among anti-ENA antibody repertoire ,the positive rate of anti-U1-nuclear ribonucleoprotein (U1-Nrnp ) antibody (52 .53% ) was the highest .Conclusion Combined detection of ANA ,anti-dsDNA antibody and anti-ENA antibodies repertoire has some clinical value of early diagnosis of SLE .%目的:探讨抗核抗体(ANA)、抗双链DNA(dsDNA)抗体及抗可溶性抗原(ENA)抗体谱对系统性红斑狼疮(SLE)的临床诊断价值。方法将158例SLE患者作为SLE组,50例健康者作为对照组。分别采用间接免疫荧光法(IIF)及免疫印迹法(IBT )对ANA、抗dsDNA抗体及抗ENA抗体谱进行检测。结果 ANA、抗dsDNA抗体及抗ENA抗体谱检测在SLE诊断中的阳性率分别为81.65%、68.35%及87.34%,均低于三者联合检测(93.04%),差异有统计学意义( P<0.05)。抗EN A抗体谱中,抗U1核糖核蛋白(U1-nRNP)抗体的阳性率(52.53%)最高。结论联合检测ANA、抗dsDNA抗体及抗ENA抗体谱对SLE的早期诊断具有一定的临床价值。

  6. Antibodies against antibodies: immunogenicity of adalimumab as a model

    NARCIS (Netherlands)

    van Schouwenburg, P.A.


    Upon repeated adalimumab exposure part of the patients start to produce ADA. The antibody response is polyclonal and consists mainly of antibodies of IgG1 and IgG4 isotype. In the majority of ADA positive patients ADA are already produced within the first 28 weeks of treatment and in part of the pat

  7. Identification of a linear epitope recognized by a monoclonal antibody directed to the heterogeneous nucleoriboprotein A2

    DEFF Research Database (Denmark)

    Tronstrøm, Julie; Dragborg, Anette H.; Hansen, Paul Robert;


    Rheumatoid arthritis (RA) is a chronic autoimmune disorder, characterized by progressive joint destruction and disability. Classical autoantibodies of RA are rheumatoid factors and citrulline antibodies. Patients positive for these autoantibodies are usually associated with a progressive disease...... to as RA33. In the absence of citrulline antibodies, RA33 antibodies have been suggested to be associated with a milder disease course. In this study we screened the reactivity of a monoclonal antibody to RA33-derived peptides by modified enzyme-linked immunosorbent assays (ELISA). Terminally truncated...

  8. Screening in liver disease

    Institute of Scientific and Technical Information of China (English)

    Paolo Del Poggio; Marzio Mazzoleni


    A disease is suitable for screening if it is common, if the target population can be identified and reached and if both a good screening test and an effective therapy are available. Of the most common liver diseases only viral hepatitis and genetic hemochromatosis partially satisfy these conditions. Hepatitis C is common, the screening test is good and the therapy eliminates the virus in half of the cases, but problems arise in the definition of the target population. In fact generalized population screening is not endorsed by international guidelines,although some recommend screening immigrants from high prevalence countries. Opportunistic screening (case finding) of individuals with classic risk factors,such as transfusion before 1992 and drug addiction,is the most frequently used strategy, but there is disagreement whether prison inmates, individuals with a history of promiscuous or traumatic sex and health care workers should be screened. In a real practice setting the performance of opportunistic screening by general practitioners is low but can be ameliorated by training programs. Screening targeted to segments of the population or mass campaigns are expensive and therefore interventions should be aimed to improve opportunistic screening and the detection skills of general practitioners. Regarding genetic hemochromatosis there is insufficient evidence for population screening, but individual physicians can decide to screen racial groups with a high prevalence of the disease, such as people in early middle age and of northern European origin. In the other cases opportunistic screening of high risk individuals should be performed, with a high level of suspicion in case of unexplained liver disease, diabetes, juvenile artropathy, sexual dysfunction and skin pigmentation.

  9. Anti IH: An antibody worth mention. (United States)

    Mohanan, Nithya; Henry, Nittin; Rafi, Aboobacker Mohamed; Innah, Susheela J


    A 72-year-old female with co-morbidities posted for surgical correction of fracture neck of femur without any history of transfusions was noted to have a hemoglobin level of 7 g/dl and packed red blood cells transfusion was ordered. Pretransfusion tests demonstrated A1B group with D positive on forward grouping. Reverse grouping showed a varying grade of agglutination with A, B, and O cells. Agglutination being stronger at 4°C. Antibody screening showed pan-agglutination, direct Coomb's test and auto control were negative. The serum reacted with adult O cells (OIadult) but not with adult Bombay cells (Oh Iadult) or O cord (Oicord) cells. A possibility of a compound cold antibody anti IH was made and A1B compatible cells were transfused to the patient. This case report illustrates anti-IH cold agglutinin with broad thermal amplitude. Uniqueness of this case report was O group incompatibility with A1B group, which was detected earlier and a catastrophic transfusion reaction being subverted.

  10. Anti IH: An antibody worth mention

    Directory of Open Access Journals (Sweden)

    Nithya Mohanan


    Full Text Available A 72-year-old female with co-morbidities posted for surgical correction of fracture neck of femur without any history of transfusions was noted to have a hemoglobin level of 7 g/dl and packed red blood cells transfusion was ordered. Pretransfusion tests demonstrated A1B group with D positive on forward grouping. Reverse grouping showed a varying grade of agglutination with A, B, and O cells. Agglutination being stronger at 4°C. Antibody screening showed pan-agglutination, direct Coomb's test and auto control were negative. The serum reacted with adult O cells (OIadult but not with adult Bombay cells (Oh Iadult or O cord (Oicord cells. A possibility of a compound cold antibody anti IH was made and A1B compatible cells were transfused to the patient. This case report illustrates anti-IH cold agglutinin with broad thermal amplitude. Uniqueness of this case report was O group incompatibility with A1B group, which was detected earlier and a catastrophic transfusion reaction being subverted.

  11. Neospora caninum antibodies in wild carnivores from Spain. (United States)

    Sobrino, R; Dubey, J P; Pabón, M; Linarez, N; Kwok, O C; Millán, J; Arnal, M C; Luco, D F; López-Gatius, F; Thulliez, P; Gortázar, C; Almería, S


    Serum samples from 251 wild carnivores from different regions of Spain were tested for antibodies to Neospora caninum by the commercial competitive screening enzyme linked immunosorbent assay (c-ELISA) and confirmed by Neospora agglutination test (NAT) and/or by indirect fluorescent antibody test (IFAT). Samples with antibodies detected by at least two serological tests were considered seropositive. Antibodies to N. caninum were found in 3.2% of 95 red foxes (Vulpes vulpes); in 21.4% of 28 wolves (Canis lupus); in 12.0% of 25 Iberian lynx (Lynx pardinus); in 16.7% of 6 European wildcats (Felis silvestris); in 6.4% of 31 Eurasian badgers (Meles meles); in 21.4% of 14 stone martens (Martes foina); in 66.7% of 3 pine martens (M. martes) and in 50% of 2 polecats (Mustela putorius). Antibodies to N. caninum in common genets (Genetta genetta) and Egyptian mongooses (Herpestes ichneumon) were only observed by c-ELISA but were not confirmed by IFAT and/or NAT. No antibodies were detected in 5 Eurasian otters (Lutra lutra) by any technique. Statistically significant differences were observed among species and among geographical areas. The highest seroprevalence of N. caninum infection was observed in the Cantabric Coastal region characterized by high humidity. To our knowledge, this is the first report of antibodies to N. caninum in free ranging wild carnivores, other than wild canids, in Europe. The existence of a possible sylvatic cycle could have important implications in both sylvatic and domestic cycles since they might influence the prevalence of infection in cattle farms in those areas.

  12. Efficient Methods To Isolate Human Monoclonal Antibodies from Memory B Cells and Plasma Cells. (United States)

    Corti, Davide; Lanzavecchia, Antonio


    In this article, we highlight the advantages of isolating human monoclonal antibodies from the human memory B cells and plasma cell repertoires by using high-throughput cellular screens. Memory B cells are immortalized with high efficiency using Epstein-Barr virus (EBV) in the presence of a toll-like receptor (TLR) agonist, while plasma cells are maintained in single-cell cultures by using interleukin 6 (IL-6) or stromal cells. In both cases, multiple parallel assays, including functional assays, can be used to identify rare cells that produce antibodies with unique properties. Using these methods, we have isolated potent and broadly neutralizing antibodies against a variety of viruses, in particular, a pan-influenza-A-neutralizing antibody and an antibody that neutralizes four different paramyxoviruses. Given the high throughput and the possibility of directly screening for function (rather than just binding), these methods are instrumental to implement a target-agnostic approach to identify the most effective antibodies and, consequently, the most promising targets for vaccine design. This approach is exemplified by the identification of unusually potent cytomegalovirus-neutralizing antibodies that led to the identification of the target, a pentameric complex that we are developing as a candidate vaccine.

  13. Pathogenic role of antiphospholipid antibodies

    NARCIS (Netherlands)

    Salmon, J. E.; de Groot, P. G.


    The antiphospholipid antibody syndrome (APS) is characterized by recurrent arterial and venous thrombosis and/or pregnancy in association with antiphospholipid (aPL) antibodies. The pathogenic mechanisms in APS that lead to in vivo injury are incompletely understood. Recent evidence suggests that AP

  14. Educational paper: Primary antibody deficiencies

    NARCIS (Netherlands)

    G.J.A. Driessen (Gertjan); M. van der Burg (Mirjam)


    textabstractPrimary antibody deficiencies (PADs) are the most common primary immunodeficiencies and are characterized by a defect in the production of normal amounts of antigen-specific antibodies. PADs represent a heterogeneous spectrum of conditions, ranging from often asymptomatic selective IgA a

  15. Targeting of Antibodies using Aptamers



    The chapter presents a methodology for the rapid selection of aptamers against antibody targets. It is a detailed account of the various methodological steps that describe the selection of aptamers, including PCR steps, buffers to be used, target immobilisation, partitioning and amplification of aptamers, clonning and sequencing, to results in high affinity and specificity ligands for the chosen target antibody.

  16. New engineered antibodies against prions (United States)

    Škrlj, Nives; Dolinar, Marko


    A number of recently developed and approved therapeutic agents based on highly specific and potent antibodies have shown the potential of antibody therapy. As the next step, antibody-based therapeutics will be bioengineered in a way that they not only bind pathogenic targets but also address other issues, including drug targeting and delivery. For antibodies that are expected to act within brain tissue, like those that are directed against the pathogenic prion protein isoform, one of the major obstacles is the blood-brain barrier which prevents efficient transfer of the antibody, even of the engineered single-chain variants. We recently demonstrated that a specific prion-specific antibody construct which was injected into the murine tail vein can be efficiently transported into brain tissue. The novelty of the work was in that the cell penetrating peptide was used as a linker connecting both specificity-determining domains of the antibody peptide, thus eliminating the need for the standard flexible linker, composed of an arrangement of three consecutive (Gly4Ser) repeats. This paves the road toward improved bioengineered antibody variants that target brain antigens. PMID:23941991

  17. Broadly Neutralizing Antibodies Display Potential for Prevention of HIV-1 Infection of Mucosal Tissue Superior to That of Nonneutralizing Antibodies. (United States)

    Cheeseman, Hannah M; Olejniczak, Natalia J; Rogers, Paul M; Evans, Abbey B; King, Deborah F L; Ziprin, Paul; Liao, Hua-Xin; Haynes, Barton F; Shattock, Robin J


    Definition of the key parameters mediating effective antibody blocking of HIV-1 acquisition within mucosal tissue may prove critical to effective vaccine development and the prophylactic use of monoclonal antibodies. Although direct antibody-mediated neutralization is highly effective against cell-free virus, antibodies targeting different sites of envelope vulnerability may display differential activity against mucosal infection. Nonneutralizing antibodies (nnAbs) may also impact mucosal transmission events through Fc-gamma receptor (FcγR)-mediated inhibition. In this study, a panel of broadly neutralizing antibodies (bnAbs) and nnAbs, including those associated with protection in the RV144 vaccine trial, were screened for the ability to block HIV-1 acquisition and replication across a range of cellular and mucosal tissue models. Neutralization potency, as determined by the TZM-bl infection assay, did not fully predict activity in mucosal tissue. CD4-binding site (CD4bs)-specific bnAbs, in particular VRC01, were consistent in blocking HIV-1 infection across all cellular and tissue models. Membrane-proximal external region (MPER) (2F5) and outer domain glycan (2G12) bnAbs were also efficient in preventing infection of mucosal tissues, while the protective efficacy of bnAbs targeting V1-V2 glycans (PG9 and PG16) was more variable. In contrast, nnAbs alone and in combinations, while active in a range of cellular assays, were poorly protective against HIV-1 infection of mucosal tissues. These data suggest that tissue resident effector cell numbers and low FcγR expression may limit the potential of nnAbs to prevent establishment of the initial foci of infection. The solid protection provided by specific bnAbs clearly demonstrates their superior potential over that of nonneutralizing antibodies for preventing HIV-1 infection at the mucosal portals of infection.

  18. Metrics for antibody therapeutics development. (United States)

    Reichert, Janice M


    A wide variety of full-size monoclonal antibodies (mAbs) and therapeutics derived from alternative antibody formats can be produced through genetic and biological engineering techniques. These molecules are now filling the preclinical and clinical pipelines of every major pharmaceutical company and many biotechnology firms. Metrics for the development of antibody therapeutics, including averages for the number of candidates entering clinical study and development phase lengths for mAbs approved in the United States, were derived from analysis of a dataset of over 600 therapeutic mAbs that entered clinical study sponsored, at least in part, by commercial firms. The results presented provide an overview of the field and context for the evaluation of on-going and prospective mAb development programs. The expansion of therapeutic antibody use through supplemental marketing approvals and the increase in the study of therapeutics derived from alternative antibody formats are discussed.

  19. ScreenOS Cookbook

    CERN Document Server

    Brunner, Stefan; Delcourt, David


    In the only book that completely covers ScreenOS, six key members of Juniper Network's ScreenOS development team help you troubleshoot secure networks using ScreenOS firewall appliances. Over 200 recipes address a wide range of security issues, provide step-by-step solutions, and include discussions of why the recipes work, so you can easily set up and keep ScreenOS systems on track. The easy-to-follow format enables you to find the topic and specific recipe you need right away.

  20. Colorectal cancer screening

    Directory of Open Access Journals (Sweden)

    Almeida Frederico Ferreira Novaes de


    Full Text Available Colorectal cancer (CRC is the third most common cancer in the world, and mortality has remained the same for the past 50 years, despite advances in diagnosis and treatment. Because significant numbers of patients present with advanced or incurable stages, patients with pre-malignant lesions (adenomatous polyps that occur as result of genetic inheritance or age should be screened, and patients with long-standing inflammatory bowel disease should undergo surveillance. There are different risk groups for CRC, as well as different screening strategies. It remains to be determined which screening protocol is the most cost-effective for each risk catagory. The objective of screening is to reduce morbidity and mortality in a target population. The purpose of this review is to analyze the results of the published CRC screening studies, with regard to the measured reduction of morbidity and mortality, due to CRC in the studied populations, following various screening procedures. The main screening techniques, used in combination or alone, include fecal occult blood tests, flexible sigmoidoscopy, and colonoscopy. Evidence from the published literature on screening methods for specific risk groups is scanty and frequently does not arise from controlled studies. Nevertheless, data from these studies, combined with recent advances in molecular genetics, certainly lead the way to greater efficacy and lower cost of CRC screening.

  1. Anti-nucleosome antibodies as a disease marker in systemic lupus erythematosus and its correlation with disease activity and other autoantibodies

    Directory of Open Access Journals (Sweden)

    Pradhan Vandana


    Full Text Available Background: Detection of anti-nucleosome antibodies (anti-nuc in patients with systemic lupus erythematosus (SLE has been well established and it is claimed that their presence is associated with disease activity. Aims: The aim of this study is to evaluate the incidence of anti-nuc antibodies and to correlate them with disease activity and its association with other autoantibodies like anti-nuclear antibodies (ANA, anti-double stranded DNA (anti-dsDNA, anti-histone antibodies (AHA, as well as autoantibodies to histone subfractions like H1, (H2A-H4 complex, H2B, and H3. Methods: This cross-sectional study included 100 SLE patients referred from the Rheumatology, Dermatology, and Nephrology Departments. SLE disease activity was evaluated by using SLE-Disease Activity Index (SLEDAI score. A patient was defined as having active SLE when the SLEDAI score was more than 5.0. Fifty normal controls were also tested as a healthy control group. Anti-nuc antibodies, anti-dsDNA, and AHA were tested by Enzyme-Linked Immunosorbent Assay (ELISA and ANA was detected by an indirect immunofluorescence test. Results: All patients studied were in an active stage of disease and were untreated, of which 44 patients had renal biopsy-proven kidney involvement, which was categorized as lupus nephritis (LN and 56 patients did not show any renal manifestations (SLE without LN. Anti-nuc antibodies were positive in 88%, anti-dsDNA in 80%, and AHA in 38% of the cases. ANA was positive in all SLE patients studied. None of the normal controls was found to be positive for these antibodies. Although a slightly higher incidence of autoantibodies were noted in LN, there was no statistical difference noted between LN and SLE without LN groups for anti-nuc and anti-dsDNA antibodies (p > 0.05. A higher incidence of autoantibodies to ANA specificities were noted in anti-nuc positive cases, but there was no statistical difference between anti-nuc positive and anti-nuc negative cases for

  2. Trypanosoma cruzi screening in Texas blood donors, 2008-2012. (United States)

    Garcia, M N; Woc-Colburn, L; Rossmann, S N; Townsend, R L; Stramer, S L; Bravo, M; Kamel, H; Beddard, R; Townsend, M; Oldham, R; Bottazzi, M E; Hotez, P J; Murray, K O


    Chagas disease is an important emerging disease in Texas that results in cardiomyopathy in about 30% of those infected with the parasite Trypanosoma cruzi. Between the years 2008 and 2012, about 1/6500 blood donors were T. cruzi antibody-confirmed positive. We found older persons and minority populations, particularly Hispanic, at highest risk for screening positive for T. cruzi antibodies during routine blood donation. Zip code analysis determined that T. cruzi is associated with poverty. Chagas disease has a significant disease burden and is a cause of substantial economic losses in Texas.

  3. Antibodies to Phospholipids and Liposomes: Binding of Antibodies to Cells (United States)


    LIPOSOMES: BINDING OF ANTIBODIES TO CELLS 12. PERSONAL AUTHOR(S) W.E. FOGLER , G. M. SWARTZ, AND C.R. ALVING 13a TYPE OF REPORT 13b. TIME COVERED 14. DATE...Elsevier BBA 73693 Antibodies to phospholipids and liposomes: binding of antibodies to cells William E. Fogler *, Glenn M. Swartz, Jr. and Carl R. Alving...Immunol. 21. Research Associateship from the U.S. National 12863-86812Hall. T. and Esser, K. (1984) 3. Immunol. 132. 2059-2063 Research Council. 13 Fogler

  4. A Study on the “Religious Imagination”in Kenzaburo Oe’ S Anti-nuclear Literature%大江健三郎反核文学中“宗教的想象力”

    Institute of Scientific and Technical Information of China (English)



    1961年首次访问“广岛”以来,大江健三郎即与“核”结下了不解之缘。他在密切关注“原子弹爆炸受害者生存状况”的同时,也在持续创作着反核题材的文学。大江的“反核文学”,以揭示“核时代人类的生存状况”为基本命题,通过政治、宗教、 SF等的想象力,创造了一个想象的世界,并在这个想象的世界中努力探索核时代人类“灵魂得以救赎”的出口。文章试图以《燃烧的绿树》《空翻》两部作品为中心,来考察大江反核题材文学中所构建的“宗教式救赎”这一体系的具体形态及本质。%Since the first visit of “Hiroshima” in 1961 , Kenzaburo Oe has been irrevocably committed to the issue of “atomic bomb” and “nuclear”.Meanwhile , He has been paying close attention to “the living conditions of the atomic bomb victims”, and has been continuing to create the literature on the theme of anti -nuclear.Oe’s anti-nuclear literature has created an imaginary world by the imagination of politics , religion , SF , etc, Which is to reveal the “the living conditions of human beings in nuclear age” and try to find the means to achieve the “salvation of soul” of the human beings .This paper attempts to clarify the nature of the“religious salvation” in Oe’ s anti-nuclear literature by analyzing the “religious imagination” in the works of The Flaming Green Tree Trilogy and Somersault .

  5. Selection of gonadotrophin surge attenuating factor phage antibodies by bioassay

    Directory of Open Access Journals (Sweden)

    Mason Helen D


    Full Text Available Abstract Background We aimed to combine the generation of "artificial" antibodies with a rat pituitary bioassay as a new strategy to overcome 20 years of difficulties in the purification of gonadotrophin surge-attenuating factor (GnSAF. Methods A synthetic single-chain antibody (Tomlinson J phage display library was bio-panned with partially purified GnSAF produced by cultured human granulosa/luteal cells. The initial screening with a simple binding immunoassay resulted in 8 clones that were further screened using our in-vitro rat monolayer bioassay for GnSAF. Initially the antibodies were screened as pooled phage forms and subsequently as individual, soluble, single-chain antibody (scAbs forms. Then, in order to improve the stability of the scAbs for immunopurification purposes, and to widen the range of labelled secondary antibodies available, these were engineered into full-length human immunoglobulins. The immunoglobulin with the highest affinity for GnSAF and a previously described rat anti-GnSAF polyclonal antiserum was then used to immunopurify bioactive GnSAF protein. The two purified preparations were electrophoresed on 1-D gels and on 7 cm 2-D gels (pH 4–7. The candidate GnSAF protein bands and spots were then excised for peptide mass mapping. Results Three of the scAbs recognised GnSAF bioactivity and subsequently one clone of the purified scAb-derived immunoglobulin demonstrated high affinity for GnSAF bioactivity, also binding the molecule in such as way as to block its bioactivity. When used for repeated immunopurification cycles and then Western blot, this antibody enabled the isolation of a GnSAF-bioactive protein band at around 66 kDa. Similar results were achieved using the rat anti-GnSAF polyclonal antiserum. The main candidate molecules identified from the immunopurified material by excision of 2-D gel protein spots was human serum albumin precursor and variants. Conclusion This study demonstrates that the combination of

  6. Mammography screening in denmark

    DEFF Research Database (Denmark)

    Vejborg, Ilse Merete Munk; Mikkelsen, Ellen Margrethe; Garne, Jens Peter


    Mammography screening is offered healthy women, and a high standard on professional and organizational level is mandatory not only in the screening programme but even in the diagnostic work-up and treatment. The main goal is to achieve a substantial reduction in disease specific mortality...

  7. Mammography screening in Denmark

    DEFF Research Database (Denmark)

    Vejborg, Ilse; Mikkelsen, Ellen Margrethe; Garne, Jens Peter


    Mammography screening is offered healthy women, and a high standard on professional and organizational level is mandatory not only in the screening programme but even in the diagnostic work-up and treatment. The main goal is to achieve a substantial reduction in disease specific mortality...

  8. Touch screens go optical

    DEFF Research Database (Denmark)

    Hanson, Steen Grüner; Jakobsen, Michael Linde; Pedersen, Henrik Chresten


    A simple optical implementation of a touch screen is made possible by disrupting the total internal reflection in a 2D waveguide.......A simple optical implementation of a touch screen is made possible by disrupting the total internal reflection in a 2D waveguide....

  9. Colorectal Cancer Screening (United States)

    ... screening tests have different risks or harms. Screening tests may cause anxiety when you are thinking about or getting ready ... is cancer when there really isn't) can cause anxiety and is usually followed by more tests (such as biopsy ), which also have risks. The ...

  10. Scoliosis Screening in Schools. (United States)

    New York State Education Dept., Albany. Div. of Pupil Personnel Services.

    The booklet outlines New York state school policy and procedures for screening students for scoliosis, lateral curvature of the spine. It is explained that screening is designed to discover spinal deformities early enough to prevent surgery. Planning aspects, including organizing a planning team for the school district, are discussed. Among…

  11. EIA screening in Denmark

    DEFF Research Database (Denmark)

    Nielsen, Eskild Holm; Christensen, Per; Kørnøv, Lone


    The article points out that EIA screening is effectively a regulatory instrument and it can be a cost-effective instrument with environmental benefits.......The article points out that EIA screening is effectively a regulatory instrument and it can be a cost-effective instrument with environmental benefits....

  12. Tabhu: tools for antibody humanization

    DEFF Research Database (Denmark)

    Olimpieri, Pier Paolo; Marcatili, Paolo; Tramontano, Anna


    Antibodies are rapidly becoming essential tools in the clinical practice, given their ability to recognize their cognate antigens with high specificity and affinity, and a high yield at reasonable costs in model animals. Unfortunately, when administered to human patients, xenogeneic antibodies can...... elicit unwanted and dangerous immunogenic responses. Antibody humanization methods are designed to produce molecules with a better safety profile still maintaining their ability to bind the antigen. This can be accomplished by grafting the non-human regions determining the antigen specificity...

  13. The isolation of novel phage display-derived human recombinant antibodies against CCR5, the major co-receptor of HIV. (United States)

    Shimoni, Moria; Herschhorn, Alon; Britan-Rosich, Yelena; Kotler, Moshe; Benhar, Itai; Hizi, Amnon


    Selecting for antibodies against specific cell-surface proteins is a difficult task due to many unrelated proteins that are expressed on the cell surface. Here, we describe a method to screen antibody-presenting phage libraries against native cell-surface proteins. We applied this method to isolate antibodies that selectively recognize CCR5, which is the major co-receptor for HIV entry (consequently, playing a pivotal role in HIV transmission and pathogenesis). We employed a phage screening strategy by using cells that co-express GFP and CCR5, along with an excess of control cells that do not express these proteins (and are otherwise identical to the CCR5-expressing cells). These control cells are intended to remove most of the phages that bind the cells nonspecifically; thus leading to an enrichment of the phages presenting anti-CCR5-specific antibodies. Subsequently, the CCR5-presenting cells were quantitatively sorted by flow cytometry, and the bound phages were eluted, amplified, and used for further successive selection rounds. Several different clones of human single-chain Fv antibodies that interact with CCR5-expressing cells were identified. The most specific monoclonal antibody was converted to a full-length IgG and bound the second extracellular loop of CCR5. The experimental approach presented herein for screening for CCR5-specific antibodies can be applicable to screen antibody-presenting phage libraries against any cell-surface expressed protein of interest.

  14. Platelet antibody detection by flow cytometry: an effective method to evaluate and give transfusional support in platelet refractoriness

    Directory of Open Access Journals (Sweden)

    Carolina Bonet Bub


    Full Text Available BACKGROUND: Immune platelet refractoriness is mainly caused by human leukocyte antigen antibodies (80-90% of cases and, to a lesser extent, by human platelet antigen antibodies. Refractoriness can be diagnosed by laboratory tests and patients should receive compatible platelet transfusions. A fast, effective and low cost antibody-screening method which detects platelet human leukocyte/platelet antigen antibodies is essential in the management of immune platelet refractoriness. OBJECTIVE: The aim of this study was to evaluate the efficiency of the flow cytometry platelet immunofluorescence test to screen for immune platelet refractoriness. METHODS: A group of prospective hematologic patients with clinically suspected platelet refractoriness treated in a referral center in Campinas, SP during July 2006 and July 2011 was enrolled in this study. Platelet antibodies were screened using the flow cytometry platelet immunofluorescence test. Anti-human leukocyte antigen antibodies were detected by commercially available methods. The sensitivity, specificity and predictive values of the immunofluorescence test were determined taking into account that the majority of antiplatelet antibodies presented human leukocyte antigen specificity. RESULTS: Seventy-six samples from 32 female and 38 male patients with a median age of 43.5 years (range: 5-84 years were analyzed. The sensitivity of the test was 86.11% and specificity 75.00% with a positive predictive value of 75.61% and a negative predictive value of 85.71%. The accuracy of the method was 80.26%. CONCLUSION: This study shows that the flow cytometry platelet immunofluorescence test has a high correlation with the anti-human leukocyte antigen antibodies. Despite a few limitations, the method seems to be efficient, fast and feasible as the initial screening for platelet antibody detection and a useful tool to crossmatch platelets for the transfusional support of patients with immune platelet refractoriness.

  15. Anti-HLA antibody profile of Turkish patients with end-stage renal disease. (United States)

    Karahan, G E; Seyhun, Y; Oguz, F; Kekik, C; Onal, E; Caliskan, Y; Bakkaloglu, H; Yazici, H; Turkmen, A; Aydin, A E; Sever, M S; Eldegez, U; Carin, M N


    Exposure to human leukocyte antigens (HLA) via blood transfusions, pregnancies, and previous transplantations can result in anti-HLA antibody production. The presence of anti-HLA antibodies in recipient sera before transplantation is an important risk factor. To demonstrate the anti-HLA antibody status of Turkish end-stage renal disease (ESRD) patients, 674 patients (mean age, 40.35 +/- 13.15 years; female/male, 328/346) were enrolled into the study. Anti-HLA antibody screening and identification tests were performed using an enzyme-linked immunosorbent assay (ELISA) method. The panel-reactive antibody (PRA)-negative group consisted of 564 (83.6%) and the PRA-positive group consisted of 110 (17.3%) patients. Of the 110 (17.3%) PRA-positive patients, 43 (6.4%) were class I (+) and class II (-); 19 (2.8%) were class I (-) and class II (+); 48 (7.1%) were both class I and II (+). The most frequent antibodies were directed against the A2 crossreactive group (CREG) and the A10 CREG with less frequent reactions against the B7 CREG, indicating antibodies to both frequent (members of A2 CREG) and relatively rare (members of A10 CREG and B7 CREG antigens). These data also suggested that some antibodies occur at greater than expected frequency because of shared epitopes. Our findings confirmed the significant correlation between female gender, pregnancy, failed graft history, long dialysis duration, and blood transfusions with PRA positivity (P < .05).

  16. A high-throughput pipeline for the production of synthetic antibodies for analysis of ribonucleoprotein complexes. (United States)

    Na, Hong; Laver, John D; Jeon, Jouhyun; Singh, Fateh; Ancevicius, Kristin; Fan, Yujie; Cao, Wen Xi; Nie, Kun; Yang, Zhenglin; Luo, Hua; Wang, Miranda; Rissland, Olivia; Westwood, J Timothy; Kim, Philip M; Smibert, Craig A; Lipshitz, Howard D; Sidhu, Sachdev S


    Post-transcriptional regulation of mRNAs plays an essential role in the control of gene expression. mRNAs are regulated in ribonucleoprotein (RNP) complexes by RNA-binding proteins (RBPs) along with associated protein and noncoding RNA (ncRNA) cofactors. A global understanding of post-transcriptional control in any cell type requires identification of the components of all of its RNP complexes. We have previously shown that these complexes can be purified by immunoprecipitation using anti-RBP synthetic antibodies produced by phage display. To develop the large number of synthetic antibodies required for a global analysis of RNP complex composition, we have established a pipeline that combines (i) a computationally aided strategy for design of antigens located outside of annotated domains, (ii) high-throughput antigen expression and purification in Escherichia coli, and (iii) high-throughput antibody selection and screening. Using this pipeline, we have produced 279 antibodies against 61 different protein components of Drosophila melanogaster RNPs. Together with those produced in our low-throughput efforts, we have a panel of 311 antibodies for 67 RNP complex proteins. Tests of a subset of our antibodies demonstrated that 89% immunoprecipitate their endogenous target from embryo lysate. This panel of antibodies will serve as a resource for global studies of RNP complexes in Drosophila. Furthermore, our high-throughput pipeline permits efficient production of synthetic antibodies against any large set of proteins.

  17. Titin antibodies in "seronegative" myasthenia gravis--A new role for an old antigen. (United States)

    Stergiou, C; Lazaridis, K; Zouvelou, V; Tzartos, J; Mantegazza, R; Antozzi, C; Andreetta, F; Evoli, A; Deymeer, F; Saruhan-Direskeneli, G; Durmus, H; Brenner, T; Vaknin, A; Berrih-Aknin, S; Behin, A; Sharshar, T; De Baets, M; Losen, M; Martinez-Martinez, P; Kleopa, K A; Zamba-Papanicolaou, E; Kyriakides, T; Kostera-Pruszczyk, A; Szczudlik, P; Szyluk, B; Lavrnic, D; Basta, I; Peric, S; Tallaksen, C; Maniaol, A; Gilhus, N E; Casasnovas Pons, C; Pitha, J; Jakubíkova, M; Hanisch, F; Bogomolovas, J; Labeit, D; Labeit, S; Tzartos, S J


    Myasthenia gravis (MG) is an autoimmune disease caused by antibodies targeting the neuromuscular junction of skeletal muscles. Triple-seronegative MG (tSN-MG, without detectable AChR, MuSK and LRP4 antibodies), which accounts for ~10% of MG patients, presents a serious gap in MG diagnosis and complicates differential diagnosis of similar disorders. Several AChR antibody positive patients (AChR-MG) also have antibodies against titin, usually detected by ELISA. We have developed a very sensitive radioimmunoprecipitation assay (RIPA) for titin antibodies, by which many previously negative samples were found positive, including several from tSN-MG patients. The validity of the RIPA results was confirmed by western blots. Using this RIPA we screened 667 MG sera from 13 countries; as expected, AChR-MG patients had the highest frequency of titin antibodies (40.9%), while MuSK-MG and LRP4-MG patients were positive in 14.6% and 16.4% respectively. Most importantly, 13.4% (50/372) of the tSN-MG patients were also titin antibody positive. None of the 121 healthy controls or the 90 myopathy patients, and only 3.6% (7/193) of other neurological disease patients were positive. We thus propose that the present titin antibody RIPA is a useful tool for serological MG diagnosis of tSN patients.

  18. Characterization of Peptide Antibodies by Epitope Mapping Using Resin-Bound and Soluble Peptides. (United States)

    Trier, Nicole Hartwig


    Characterization of peptide antibodies through identification of their target epitopes is of utmost importance. Understanding antibody specificity at the amino acid level provides the key to understand the specific interaction between antibodies and their epitopes and their use as research and diagnostic tools as well as therapeutic agents. This chapter describes a straightforward strategy for mapping of continuous peptide antibody epitopes using resin-bound and soluble peptides. The approach combines three different types of peptide sets for full characterization of peptide antibodies: (1) overlapping peptides, used to locate antigenic regions; (2) truncated peptides, used to identify the minimal peptide length required for antibody binding; and (3) substituted peptides, used to identify the key residues important for antibody binding and to determine the specific contribution of key residues. For initial screening resin-bound peptides are used for epitope estimation, while soluble peptides subsequently are used for fine mapping. The combination of resin-bound peptides and soluble peptides for epitope mapping provides a time-sparing and straightforward approach for characterization of peptide antibodies.

  19. A Model System for Concurrent Detection of Antigen and Antibody Based on Immunological Fluorescent Method

    Directory of Open Access Journals (Sweden)

    Yuan-Cheng Cao


    Full Text Available This paper describes a combined antigen/antibody immunoassay implemented in a 96-well plate using fluorescent spectroscopic method. First, goat anti-human IgG was used to capture human IgG (model antigen; goat anti-human IgG (Cy3 or FITC was used to detect the model antigen; a saturating level of model antigen was then added followed by unlabelled goat anti-human IgG (model antibody; finally, Cy3 labelled rabbit anti-goat IgG was used to detect the model antibody. Two approaches were applied to the concomitant assay to analyze the feasibility. The first approach applied FITC and Cy3 when both targets were present at the same time, resulting in 50 ng/mL of the antibody detection limit and 10 ng/mL of antigen detection limit in the quantitative measurements of target concentration, taking the consideration of FRET efficiency of 68% between donor and acceptor. The sequential approach tended to lower the signal/noise (S/N ratio and the detection of the model antigen (lower than 1 ng/mL had better sensitivity than the model antibody (lower than 50 ng/mL. This combined antigen/antibody method might be useful for combined detection of antigens and antibodies. It will be helpful to screen for both antigen and antibody particularly in the situations of the multiserotype and high-frequency mutant virus infections.

  20. Phage-display libraries of murine and human antibody Fab fragments

    DEFF Research Database (Denmark)

    Engberg, J; Andersen, P S; Nielsen, L K


    We provide efficient and detailed procedures for construction, expression, and screening of comprehensive libraries of murine or human antibody Fab fragments displayed on the surface of filamentous phage. In addition, protocols for producing and using ultra-electrocompetent cells, for producing Fab...

  1. Development of a Highly Protective Combination Monoclonal Antibody Therapy against Chikungunya Virus

    NARCIS (Netherlands)

    Pal, Pankaj; Dowd, Kimberly A.; Brien, James D.; Edeling, Melissa A.; Gorlatov, Sergey; Johnson, Syd; Lee, Iris; Akahata, Wataru; Nabel, Gary J.; Richter, Mareike K. S.; Smit, Jolanda M.; Fremont, Daved H.; Pierson, Theodore C.; Heise, Mark T.; Diamond, Michael S.


    Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes global epidemics of a debilitating polyarthritis in humans. As there is a pressing need for the development of therapeutic agents, we screened 230 new mouse anti-CHIKV monoclonal antibodies (MAbs) for their ability to inhibit

  2. Survey of citrus tristeza virus populations in Central California that react with MCA13 monoclonal antibody (United States)

    The Citrus Pest Detection Program (CPDP) of the Central California Tristeza Eradication Agency monitors Citrus tristeza virus (CTV) in Central California. MCA13 is a severe strain discriminating monoclonal antibody used to screen for potentially virulent CTV isolates. MCA13-reactive CTV isolates are...

  3. Broadly Neutralizing Alphavirus Antibodies Bind an Epitope on E2 and Inhibit Entry and Egress

    NARCIS (Netherlands)

    Fox, Julie M.; Long, Feng; Edeling, Melissa A.; Lin, Hueylie; van Duijl-Richter, Mareike K. S.; Fong, Rachel H.; Kahle, Kristen M.; Smit, Jolanda M.; Jin, Jing; Simmons, Graham; Doranz, Benjamin J.; Crowe, James E.; Fremont, Daved H.; Rossmann, Michael G.; Diamond, Michael S.


    We screened a panel of mouse and human monoclonal antibodies (MAbs) against chikungunya virus and identified several with inhibitory activity against multiple alphaviruses. Passive transfer of broadly neutralizing MAbs protected mice against infection by chikungunya, Mayaro, and O'nyong'nyong alphav

  4. Screening detected celiac disease in children with type 1 diabetes mellitus : Effect on the clinical course - (A case control study)

    NARCIS (Netherlands)

    Rami, B; Sumnik, Z; Schober, E; Waldhor, T; Battelino, T; Bratanic, N; Kurti, K; Lebl, J; Limbert, C; Madacsy, L; Odink, RJH; Paskova, M; Soltesz, G


    Objective: To investigate clinical and metabolic characteristics of diabetic children with screening detected celiac disease in a multicenter case-control study. Methods: Cases: 98 diabetic patients were diagnosed as having silent celiac disease by screening with endomysial antibodies and subsequent

  5. Screening detected celiac disease in children with type 1 diabetes mellitus: Effect on the clinical course - (A case control study)

    NARCIS (Netherlands)

    Rami, B.; Sumnik, Z.; Schober, E.; Waldhor, T.; Battelino, T.; Bratanic, N.; Kurti, K.; Lebl, J.; Limbert, C.; Madacsy, L.; Odink, R.J.H.; Paskova, M.; Soltesz, G.


    Objective: To investigate clinical and metabolic characteristics of diabetic children with screening detected celiac disease in a multicenter case-control study. Methods: Cases: 98 diabetic patients were diagnosed as having silent celiac disease by screening with endomysial antibodies and subsequent

  6. Barriers to cancer screening. (United States)

    Womeodu, R J; Bailey, J E


    Many barriers to cancer screening have been summarized and discussed. Barriers have been documented in all patient populations, but some groups such as ethnic minorities and the elderly face unique barriers. The barriers to cancer screening, are multifactorial, but much of the responsibility for change must lie with health care providers and the health care delivery industr