Application and development of a TaqMan real-time PCR for detecting infectious spleen and kidney necrosis virus in Siniperca chuatsi

文献类型： 外文期刊

第一作者： Lin, Qiang

作者： Lin, Qiang;Fu, Xiaozhe;Liu, Lihui;Liang, Hongru;Guo, Huizhi;Yin, Shuwen;Huang, Zhibin;Li, Ningqiu;Kumaresan, Venkatesh

作者机构：

关键词： Chinese perch;ISKNV;TaqMan real-time PCR

期刊名称：MICROBIAL PATHOGENESIS （ 影响因子：3.738； 五年影响因子：3.663 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Infectious spleen and kidney necrosis virus (ISKNV) is one of the major epidemiological agents that had caused great economic loss in Chinese perch (Siniperca chuatsi). In this study, a specific TaqMan real-time PCR was developed using a pair of primers and a TaqMan probe specific to the ORF007 gene of ISKNV to rapidly detect ISKNV copies in Chinese perch samples. This assay was optimized to produce linearity from 8.75 x 10(8) to 8.75 x 10(1) copies in standard curve with an efficiency of 98% and a R-2 value of 0.9999. Moreover, the minimum detection limit of this assay was 10,000 times more sensitive than that of conventional PCR method. The coefficients of variation of intra- and inter-assay repeatability were less than 2.4% and 3.3%, respectively. The viral distribution in different tissues of diseased Chinese perch was evaluated by TaqMan real-time PCR method and the highest level of viral copies was detected in spleen. Among the 76 diseased Chinese perch clinical samples, 35 and 29 were positive samples based on the TaqMan real-time PCR and conventional PCR methods, respectively, indicating that the TaqMan real-time PCR was more sensitive than conventional PCR. Therefore, the TaqMan real-time PCR should be a useful tool for the early surveillance and quantitation of ISKNV. (C) 2017 Elsevier Ltd. All rights reserved.

分类号： S85

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