Development of a rapid and specific loop-mediated isothermal amplification detection method that targets Marek's disease virus meq gene

文献类型： 外文期刊

第一作者： Wei, Xiuying

作者： Wei, Xiuying;Shi, Xingming;Zhao, Yan;Zhang, Jing;Wang, Mei;Liu, Changjun;Cui, Hongyu;Hu, Shunlei;Quan, Yanming;Chen, Hongyan;Wang, Yunfeng;Wei, Xiuying

作者机构：

关键词： Loop-mediated isothermal amplification;Marek's disease virus;MDV detection;meq gene

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2012 年 183 卷 2 期

页码：

收录情况： SCI

摘要： A rapid, sensitive and specific loop-mediated isothermal amplification (LAMP) method was developed and evaluated for the detection of Marek's disease virus (MDV) by amplification of conserved MDV meq gene sequences. LAMP is an innovative technique that allows the rapid detection of targeted nucleic acid sequences under isothermal conditions without the need for complex instrumentation. In this study, meq gene sequences were amplified successfully from different MDV strains by LAMP within 60 min and no cross-reactivity was observed in a panel of related viruses that were associated with diseases of chickens. The detection limit of LAMP was 3.2 copies/million cells compared with 320 copies/million cells required for conventional PCR. Positive detection rates were assessed using either LAMP or PCR by examination of feather follicles that were collected from chickens infected experimentally with either strain J-1 (n=20) or strain Md5 (n=17), In addition to these samples, three isolates that were suspected to have been infected in the clinic were also tested. Results showed that the positive detection rate for LAMP was 95% (38/40), compared with 87.5% (35/40) and 90% (38/40) for strains J-1 and Md5 by PCR, respectively. These results indicated that the LAMP assay was more sensitive, rapid and specific than conventional PCR for the detection of MDV. This easy-to-perform technique will be useful for the detection of MDV and will aid in the establishment of disease control protocols. (C) 2012 Elsevier B.V. All rights reserved.

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