Detection and quantification of Aeromonas schubertii in Channa maculata by TaqMan MGB probe fluorescence real-time quantitative PCR

文献类型： 外文期刊

第一作者： Liu, Chun

作者： Liu, Chun;Lin, Li;Liu, Chun;Cao, Jizhen Z.;Zhang, De-Feng;Chang, Ou-Qin;Li, Kaibin;Wang, Fang;Shi, Cun-Bin;Jiang, Lan;Wang, Qing;Guo, Yanming M.;Lin, Li

作者机构：

关键词： Aeromonas schubertii; bacterial load; Channidae; TaqMan MGB real-time PCR

期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.767； 五年影响因子：2.689 ）

ISSN： 0140-7775

年卷期： 2019 年 42 卷 1 期

页码：

收录情况： SCI

摘要： Aeromonas schubertii is a major epidemiological agent that threatens cultured snakeheads (Channidae) and has caused great economic losses in fish-farming industries in China in recent years. In present study, a specific TaqMan minor groove binder (MGB) probe fluorescence real-time quantitative PCR (qPCR) assay was developed to rapidly detect and quantify A. schubertii. A pair of qPCR primers and a TaqMan MGB probe were selected from the rpoD gene, which were shown to be specific for A. schubertii. A high correlation coefficient (R-2 = 0.9998) in a standard curve with a 103% efficiency was obtained. Moreover, the qPCR method's detection limit was as low as 18 copies/mu l, which was 100 times more sensitive than that of conventional PCR. The detection results for the A. schubertii in pond water and fish tissue were consistent with those of the viable counts. Bacterial load changes detected by qPCR in different tissues of snakeheads infected with A. schubertii showed that the gills and intestines may be the entry for A. schubertii, and the spleen and kidney are major sites for A. schubertii replication. The established method in present study should be a useful tool for the early surveillance and quantitation of A. schubertii.

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