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Development and partial validation of a recombinant E2-based indirect ELISA for detection of specific IgM antibody responses against classical swine fever virus

文献类型： 外文期刊

作者： Li, Wenliang 1 ; Mao, Li 1 ; Yang, Leilei 1 ; Zhou, Bin 2 ; Jiang, Jieyuan 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Med, Natl Ctr Engn Res Vet Bioprod, Key Lab Vet Biol Engn & Technol,Minist Agr, Nanjing 210014, Jiangsu, Peoples R China

2.Nanjing Agr Univ, Coll Vet Med, Nanjing 210095, Jiangsu, Peoples R China

关键词： CSFV;E2 protein;IgM antibody;ELISA

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2013 年 191 卷 1 期

页码：

收录情况： SCI

摘要： Detecting classical swine fever virus specific antibody responses is critical for prevention and control of CSF. In this study, a Delta E2-based indirect ELISA was developed to detect specific IgM antibodies against CSFV. The optimized conditions that were determined experimentally are: a Delta E2 antigen concentration of 0.5 mu g/ml, a serum sample dilution of 1/100 incubated at 37 degrees C for 1.5 h, and a HRP conjugated rabbit anti-pig IgM dilution of 1/50,000 incubated at 37 degrees C for 1 h. Three hundred clinical sera were tested with Delta E2-IgM-ELISA and IDEXX ELISA and the positive rates were 77.3% (232/300) and 71.7% (215/300), respectively. Concordance rate between them was 80.3% (241/300). The 59 inconsistent sera were tested further: among the 21 IDEXX ELISA +/Delta E2-IgM-ELISA - and 38 IDEXX ELISA +/Delta E2-IgM-ELISA - samples, 17 and 24 were determined positive by virus neutralization test; 15 and 25 were tested positive by Delta E2-IgG-ELISA, respectively. In addition, the E2-specific IgM antibody response in 15 vaccinated piglets could be detected 2 weeks post-vaccination and earlier than specific IgG antibody. It increased regularly and reached high levels by 6 weeks post-vaccination. The Delta E2-IgM-ELISA could be used for clinical detection and for exploring the kinetics of IgM antibody response. (C) 2013 Published by Elsevier B.V.

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