Development of TaqMan real-time PCR assay for detection and quantitation of reticuloendotheliosis virus

文献类型： 外文期刊

第一作者： Li, Kai

作者： Li, Kai;Gao, Honglei;Gao, Li;Qi, Xiaole;Qin, Liting;Gao, Yulong;Xu, Yanwei;Wang, Xiaomei

作者机构：

关键词： Reticuloendotheliosis virus (REV);Real-time PCR;Proviral load;Detection

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 179 卷 2 期

页码：

收录情况： SCI

摘要： A highly sensitive real-time PCR method was developed in this study for reticuloendotheliosis virus (REV) detection and quantitation. The real-time PCR method, with a minimum detection limit of 10 proviral DNA copies, was 100 times more sensitive than the conventional PCR. It was also shown to be highly specific, as no positive signals were detected for other common avian DNA viruses. The coefficients of variation of intra- and inter-assay reproducibility were both less than 2%. The chicken beta-actin gene was co-amplified and used as the internal control to monitor the efficiency of DNA extraction and PCR amplification. Specific pathogen free chickens were infected with REV at different ages and the blood was detected with the real-time PCR method. High levels of proviral DNA were detected in the blood of REV-infected chickens during the experiment and chickens infected early had higher proviral loads from 2 weeks post-infection compared with late infected chickens. This study provides an excellent research and diagnostic tool that can be used for REV detection and quantitation. (C) 2011 Elsevier B.V. All rights reserved.

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