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#27661084   2016/09/23 Save this To Up

Simultaneous Quantification of Anticardiolipin IgG and IgM by Time Resolved Fluoroimmunoassay.

The autoimmune disease antiphospholipid syndrome (APS) is characterized by the presence of anticardiolipin antibodies (aCL), along with anti-β2-glycoprotein I (β2GPI) antibodies and lupus anticoagulant (LA). In this study, we developed a time-resolved fluoroimmunoassay (TRFIA) system for simultaneous quantification of aCL IgG and IgM. A 96-well microtiter plate precoated with the complex of cardiolipin from bovine heart and bovine β2GPI was incubated with the anticardiolipin IgG and IgM standard substance or serum, and the conjugate of Eu3+-labeled anti-human IgG and Sm3+-labeled anti-human IgM was pipetted to the wells to form a tipical double-antibody-sandwich immunoreactions; finally the fluorescent intensity of Eu3+ and Sm3+ was detected to reflect the quantity of anticardiolipin IgG and IgM. This assay showed a good relationship between fluorescence intensities and the concentration of anticardiolipin antibody(aCL) IgG and IgM, with a low-end sensitivity of 0.1 U/ml for IgG and 0.1 U/ml for IgM, respectively. The intra- and inter-assay coefficients of variation (CV) of the calibrators was 3.0% and 4.51% for IgG, and 2.76% and 4.45% for IgM. The average recovery was 100.38% for aCL IgG and 100.45% for aCL IgM. For serum samples, the results of our method showed a good correlation with those obtained with ELISA kit. Simultaneous detection of aCL-IgG and aCL-IgM in the same reaction well can optimize assay performance by avoiding potential influence of different reaction conditions-timing, and well-to-well difference in concentration and characteristics of cardiolipin antigen. The results of a combo aCL-IgG and aCL-IgM assay for the same sample are more consistent and more reliable. This dual-label time-resolved fluoroimmunoassay is sensitive for detecting aCL IgG and IgM across a wide concentration range with stable reagents and may assist in the clinical diagnosis of antiphospholipid syndrome.

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#24659029   2014/07/21 Save this To Up

A new Eu(3+)-labeled method for anticardiolipin antibody IgM.

The anticardiolipin antibodies (aCL) test has become a laboratory standard for the clinical diagnosis of antiphospholipid syndrome (APS). To better the quantitative detection of aCL-IgM so as to classify patients correctly and timely as APS positive, we established herein a new immunoassay based on a time-resolved fluoroimmunoassay (TRFIA).

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MOUSE ANTI HUMAN CD15, Pr MOUSE ANTI HUMAN CD15, Pr MOUSE ANTI BOVINE ROTAVIR Human IgM antibody, Monoc FITC antibody, Monoclonal ACE antibody, Monoclonal Adenovirus antibody, Mono CA 50 antibody, Monoclona Campylobacter jejuni anti DNA antibody, Monoclonal Gram Negative Endotoxin a HBsAg antibody, Monoclona

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#23656246   2013/05/09 Save this To Up

Detection of anticardiolipin antibody igm by sm(3+)-labeled time-resolved fluoroimmunoassay.

In an effort to improve the quantitative detection of aCL IgM, we develop a new immunoassay to improve aCL IgM detection based on TRFIA using the complex of cardiolipin plus bovine β2GPI as antigen and Sm(3+)-labeled rabbit anti-human IgM as conjugate. The precision, sensitivity, specificity, coefficient of recovery, and stability of the assay were evaluated and comparison with the classical ELISA was also made. The aCL IgM TRFIA kit we established had a wider detectable range than commercial ELISA ones when diluted a specimen with strong positive from 1:2.5-1:40960. We observed that for the established TRFIA kit there was a good liner range within 1:2.5-1:40960, whereas it was within 1:20-1:1280 when using ELISA kits. The intraassay precision rate and the interassay precision rate were <5% for 3 different concentrations. The sensitivity was 0.1MPL U/mL and the clinical diagnostic specificity was 98%. Average recovery rate was 101.13%. The established assay kit also behaved better in stability. Additionally, the immunoassay we established correlated well with the ELISA and the correlation coefficient was 0.956. We thus conclude that the TRFIA we developed for aCL IgM detection gives promise to a more sensitivity and reliable diagnosis of APS and has potential value for large-scale screening programs.

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#22616022   2012/05/22 Save this To Up

Detection, isolation and confirmation of Crimean-Congo hemorrhagic fever virus in human, ticks and animals in Ahmadabad, India, 2010-2011.

In January 2011, human cases with hemorrhagic manifestations in the hospital staff were reported from a tertiary care hospital in Ahmadabad, India. This paper reports a detailed epidemiological investigation of nosocomial outbreak from the affected area of Ahmadabad, Gujarat, India.

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#19912047   2009/11/16 Save this To Up

Detection of myelin autoantibodies: evaluation of an assay system for diagnosis of multiple sclerosis in differentiation from other central nervous system diseases.

Multiple sclerosis (MS) is a frequent and often severe autoimmune disease of the central nervous system. We describe a newly developed enzyme-linked immunosorbent assay (ELISA)-based test system for the assessment of neuronal autoantibodies in serum and cerebrospinal fluid (CSF). This tool could help define autoimmune status and thus be a potential means of therapeutic surveillance.

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#16697531   2006/08/21 Save this To Up

Neospora caninum antibodies in commercial fetal bovine serum.

The protozoan parasite Neospora caninum is a major cause of abortion in cattle throughout the world. In the process of propagating Neospora in vitro and producing specific antibodies for development of diagnostic assays in the food supply, our laboratory identified the presence of bovine antibodies to N. caninum in fetal bovine sera. The sera were produced commercially and preferentially recommended for tissue culture use and monoclonal antibody production. Seventeen different fetal bovine serum samples of different grades and from four different companies were examined for the presence of total IgG, IgG1, IgG2, and IgM specific for N. caninum. All of the tested serum samples recognized N. caninum specific bands on Western blot. Low IgG serum also recognized these antigens but with lower intensity. Antibody response was also evaluated using a commercially available ELISA kit for N. caninum.

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#12726929   2003/05/02 Save this To Up

Evaluation of newly developed ELISA using "MESACUP-2 test mitochondrial M2" kit for the diagnosis of primary biliary cirrhosis.

An enzyme-linked immunosorbent assay (ELISA) using MESACUP-2 Test Mitochondria M2 kit (new-M2 ELISA) has recently become commercially available. The aim of this study was to evaluate the clinical utility of this newly developed ELISA for the diagnosis of primary biliary cirrhosis (PBC).

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#11982703   2002/05/01 Save this To Up

Automated enzymatic mitochondrial antibody assay for the diagnosis of primary biliary cirrhosis: applications of a routine diagnostic tool for the detection of antimitochondrial antibodies.

An automated enzymatic mitochondrial antibody assay (EMA) kit for the diagnosis of primary biliary cirrhosis (PBC) has become commercially available recently. The aim of this study was to assess the clinical utility of the enzyme inhibition assay using this EMA kit for the diagnosis of PBC.

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