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J Virol Methods. 2013 Jul;191(1):63-8. doi: 10.1016/j.jviromet.2013.03.003. Epub 2013 Mar 14.

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

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  • 1Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, China.

Abstract

Detecting classical swine fever virus specific antibody responses is critical for prevention and control of CSF. In this study, a ΔE2-based indirect ELISA was developed to detect specific IgM antibodies against CSFV. The optimized conditions that were determined experimentally are: a ΔE2 antigen concentration of 0.5 μg/ml, a serum sample dilution of 1/100 incubated at 37°C for 1.5 h, and a HRP conjugated rabbit anti-pig IgM dilution of 1/50,000 incubated at 37°C for 1 h. Three hundred clinical sera were tested with ΔE2-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 +/ΔE2-IgM-ELISA - and 38 IDEXX ELISA +/ΔE2-IgM-ELISA - samples, 17 and 24 were determined positive by virus neutralization test; 15 and 25 were tested positive by ΔE2-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 ΔE2-IgM-ELISA could be used for clinical detection and for exploring the kinetics of IgM antibody response.

PMID:
23500647
DOI:
10.1016/j.jviromet.2013.03.003
[PubMed - indexed for MEDLINE]
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