Development and application of an indirect enzyme-linked immunosorbent assay for serological survey of Japanese encephalitis virus infection in dogs
Development and application of an indirect enzyme-linked immunosorbent assay for serological survey of Japanese encephalitis virus infection in dogs
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DOI：
10.1016/j.jviromet.2012.09.022
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Maeda, Ken
中科院分区：
文献类型：
--
作者：
Shimoda, Hiroshi;Inthong, Natnaree;Maeda, Ken
日本脑炎病毒（JEV）可引起人和马严重的急性脑炎。虽然狗是评估人类感染JEV风险的良好哨兵，但病毒中和试验一直是测量狗体内JEV抗体水平的唯一方法。本研究以纯化的病毒颗粒为抗原，建立了间接酶联免疫吸附试验（ELISA），用于犬乙型脑炎病毒感染的血清学检测。在实验接种JEV的狗中，ELISA在感染后3天检测到抗JEV IgM，IgM水平在感染后7天达到峰值。抗JEV IgG在感染后14天开始检测，21-28天达高峰。病毒中和滴度与通过ELISA测量的抗JEV免疫球蛋白相关。为了测试新的检测方法的实用性，在日本九州的102只狗中，JEV感染的血清阳性率通过IgG ELISA和病毒中和进行了检查。IgG ELISA法与病毒中和法的相关系数为0.813（p < 0.001），敏感性为82%，特异性为98%。IgG ELISA被用来调查狗在曼谷，泰国和51%的这些狗被发现为JEV血清阳性。这些数据表明，在泰国首都，感染JEV的风险仍然很高。(C)2012爱思唯尔有限公司版权所有。
Japanese encephalitis virus (JEV) causes serious acute encephalitis in humans and horses. Although dogs are good sentinels for assessing the risk of JEV infection to humans, a virus neutralization test has been the only method available for measuring the levels of JEV antibody in dogs. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) using purified viral particles as an antigen, was developed for serological survey of JEV infection in dogs. In dogs inoculated experimentally with JEV, the ELISA detected anti-JEV IgM 3 days after infection, with IgM levels peaking 7 days after infection. Anti-JEV IgG was detected 14 days after infection and peaked on 21-28 days after infection. Virus neutralization titers correlated with anti-JEV immunoglobulins measured by the ELISA. To test the utility of the new assay, the seroprevalence of JEV infection among 102 dogs in Kyushu, Japan, was examined by IgG ELISA and by virus neutralization. The correlation coefficient between the IgG ELISA and virus neutralization was 0.813 (p < 0.001): comparison of the IgG ELISA and virus neutralization showed a sensitivity and specificity of 82% and 98%, respectively. The IgG ELISA was used to survey dogs in Bangkok, Thailand and 51% of these dogs were found seropositive for JEV. These data suggest that in the capital city of Thailand, the risk of infection with JEV remains high. (C) 2012 Elsevier B.V. All rights reserved.