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Open Access Highly Accessed Methodology

Development and application of an antigen capture ELISA assay for diagnosis of Japanese encephalitis virus in swine, human and mosquito

Li Mei12, Peng Wu3, Jing Ye12, Guangping Gao4, Lin Shao12, Shaomei Huang12, Yaoming Li12, Xiaohong Yang12, Huanchun Chen12 and Shengbo Cao12*

Author Affiliations

1 State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei 430070, China

2 Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China

3 Department of Clinical Laboratory, Taihe Hospital, Hubei University of Medicine, 16 Shiyan, Shiyan 442000, China

4 Department of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066600, China

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Virology Journal 2012, 9:4  doi:10.1186/1743-422X-9-4

Published: 6 January 2012

Abstract

Background

Japanese encephalitis (JE) is a serious zoonosis caused by the Japanese encephalitis virus (JEV) which is a mosquito-borne pathogen of the family Flavivirus. However, the application of several developed laboratory methods for the detection of JEV antigens or antibodies are limited by their requirements of laboratory operations, skilled technicians and special facilities.

Results

To develop a method for detecting JEV antigen in swine, human, mosquito and other clinical specimens specifically, conveniently and effectively, an antigen capture enzyme-linked immunosorbent assay (ELISA) was established in this study. Sensitivity, specificity, repeatability and stability of the developed method were evaluated, and 60 clinical samples were tested in this study. The results demonstrated that the antigen capture ELISA was capable in detecting JEV antigen with high sensitivity and specificity compared with conventional methods. 14 samples showed the positive result with coincidence rate of 70%, and 46 displayed negative result with coincidence rate of 100% as compared to that of reverse transcription-polymerase chain reaction (RT-PCR).

Conclusions

The developed ELISA assay provides a convenient and specific method for the large-scale determination of JEV antigen in infected swine, human and mosquito samples with high sensitivity and specificity.

Keywords:
Japanese encephalitis; Japanese encephalitis virus; Monoclonal antibody; Polyclonal antibody; Antigen capture ELISA