Development and validation of a prokaryotically expressed foot-and-mouth disease virus non-structural protein 2C'3AB-based immunochromatographic strip to differentiate between infected and vaccinated animals
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- 作者：Lei Wu (1)
Tao Jiang (1)
Zeng-Jun Lu (1)
Ya-Min Yang (1)
Pu Sun (1)
Zhong Liang (1)
Dong Li (1)
Yuan-Fang Fu (1)
Yi-Mei Cao (1)
Xiang-Tao Liu (1)
Zai-Xin Liu (1) - 关键词：an immunochromatographic strip ; foot ; and ; mouth disease virus ; colloidal gold ; labeled 2C'3AB antigen ; prokaryotic expression ; development ; validation
- 刊名：Virology Journal
- 出版年：2011
- 出版时间：December 2011
- 年：2011
- 卷：8
- 期：1
- 全文大小：1329KB
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Tao Jiang (1)
Zeng-Jun Lu (1)
Ya-Min Yang (1)
Pu Sun (1)
Zhong Liang (1)
Dong Li (1)
Yuan-Fang Fu (1)
Yi-Mei Cao (1)
Xiang-Tao Liu (1)
Zai-Xin Liu (1)
1. State Key Laboratory of Veterinary Etiologic Biology, National Foot-and-Mouth Disease Reference Laboratory of China, Key laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, Gansu, China - ISSN：1743-422X
文摘
Background Foot-and-mouth disease (FMD) is an extremely contagious viral disease of cattle, pigs, sheep, goats, and many cloven-hoofed wild animals. FMDV serotypes O and Asia 1 have circulated separately in China during the last fifty years, and eliminating infected animals and vaccination are the main policies to prevent and control FMD. Antibodies to NSPs exist in infected animals, and were utilized to differentiate between infected and vaccinated animals. The reliability of detection of 3AB or 3ABC antibodies is higher than that of other NSPs. The test of 3AB is still credible because 3C protein's immunogenicity is the weakest. The 2C protein, immediately N-terminal of 3AB, was used to differentiate between infected and vaccinated animals. The use of the immunochromatographic strip is facile for clinical laboratories lacking specialized equipment and for rapid field diagnosis. Results In this study, an immunochromatographic strip with non-structural protein (NSP) 2C'3AB was developed and validated to differentiate foot-and-mouth disease infected from vaccinated animals. A part of N-terminal of 2C protein gene and whole 3AB gene were connected and prokaryotically expressed as the antigens labeled with colloidal gold was used as the detector, the 2C'3AB protein and rabbits anti-2C'3AB antibodies were blotted on the nitrocellulose(NC) membrane for the test and control lines, respectively. 387 serum samples were collected to evaluate the characteristics of the strip in comparison with existing commercial 3ABC antibody ELISA kit. The coincidence rate of pigs negative serum, pigs vaccinated serum, pigs infected serum was 100%, 97.2%, 95.0%, respectively. The coincidence rate of cattle negative serum, cattle vaccinated serum, cattle infected serum was 100%, 96.7%, 98.0%, respectively. The coincidence rate of sheep negative serum, sheep infected serum was 97.6%, 96.3%, respectively. The strip was shown to be of high specificity and sensitivity, good repeatability and stability. Conclusion These data suggest that the immunochromatographic strip is a useful tool for rapid on-site diagnosing animals infected foot-and-mouth disease virus.