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Published in: Archives of Virology 6/2019

01-06-2019 | Encephalitis | Original Article

Development and application of a monoclonal-antibody-based blocking ELISA for detection of Japanese encephalitis virus NS1 antibodies in swine

Authors: Dengyuan Zhou, Chao Pei, Kelu Yang, Jing Ye, Shengfeng Wan, Qiuyan Li, Luping Zhang, Huanchun Chen, Shengbo Cao, Yunfeng Song

Published in: Archives of Virology | Issue 6/2019

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Abstract

Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted by Culex mosquitoes and is the leading cause of viral encephalitis in humans. JEV infection of swine, which are the main amplifying hosts for JEV, can cause reproductive failure in sows; in boars it can cause testitis and infertility. The prevalence of JEV in swine is a continuous threat to human health. A practical diagnostic method for monitoring JEV infection in swine herds is essential for control of the disease in both swine and humans. Here, we have identified a high-affinity anti-JEV NS1 monoclonal antibody (mAb) by indirect ELISA and utilized it for the development of a blocking ELISA (bELISA). The optimal NS1 protein coating concentration (2 μg/mL) and mAb working concentration (1 μg/mL) were determined by checkerboard titration. One hundred ten JEV-antibody-negative serum samples were used to establish 34.03% inhibition as the cutoff value for a negative result. By the bELISA, seroconversion in 80% of newly JEV-vaccinated pigs was detected by 7 days post-immunization, while by the commercial envelope-protein-based iELISA, seroconversion was detected in 20% of the newly vaccinated pigs. We found 98.7% agreement between the bELISA and the commercial iELISA when we tested 157 field samples using both methods. From an epidemiological survey of swine serum collected between 2014 and 2016, we found that the average JEV seropositive rate in unvaccinated commodity pigs was 8.1%, and in vaccinated boars and sows, it was 67.6%.
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Metadata
Title
Development and application of a monoclonal-antibody-based blocking ELISA for detection of Japanese encephalitis virus NS1 antibodies in swine
Authors
Dengyuan Zhou
Chao Pei
Kelu Yang
Jing Ye
Shengfeng Wan
Qiuyan Li
Luping Zhang
Huanchun Chen
Shengbo Cao
Yunfeng Song
Publication date
01-06-2019
Publisher
Springer Vienna
Keyword
Encephalitis
Published in
Archives of Virology / Issue 6/2019
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-019-04218-9

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