Development of a rapid and sensitive reverse transcription real-time quantitative PCR assay for detection and quantification of grass carp reovirus II

文献类型： 外文期刊

第一作者： Li, Jiahao

作者： Li, Jiahao;Wang, Yajun;Wang, Yingying;Li, Yingying;Shi, Cunbin;Mo, Xubing;Wang, Qing;Yin, Jiyuan;Wu, Huiliang;Xu, Wei;Wang, Hao;Bergmann, Sven M.;Wang, Qing;Yin, Jiyuan

作者机构：

关键词： GCRV-II; Quantitative analysis; RT-qPCR; Detection

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：3.1； 五年影响因子：2.4 ）

ISSN： 0166-0934

年卷期： 2023 年 312 卷

页码：

收录情况： SCI

摘要： Hemorrhagic disease of grass carp, which is induced by grass carp reovirus II (GCRV-II), leads to mass mortality in grass carp culture and causes enormous economic loss. However, there is currently no quantitative analysis method for the detection of GCRV-II, which is greatly restricted the etiological and epidemiological study of the disease. In this study a reverse transcription TaqMan PCR (RT-qPCR) assay was developed for the quantitative detection of GCRV-II. The probe and primers targeted location is the segment 6 (S6) region of the GCRV-II genome which is highly conserved. Standard curves were drawn and criteria were confirmed after the deter-mination of the optimum reaction conditions. The species-specific assay showed that the method is highly specific and has no cross reactions with other pathogens. The assay was sufficiently sensitive to detect as low as 10 copies of virus RNA. Moreover, the method has a very good repeatability for batches and inter-batches sample detection. Then the method was applied to detect the virus in tissue samples from clinically infected grass carp, compared with conventional RT-seminested PCR, the RT-qPCR represents a specific value for detection rate of positive samples. In summary, the RT-qPCR was applied and achieved high sensitivity and specificity for GCRV-II detection.

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