Development and application of a sensitive droplet digital PCR-based method to detect tilapia parvovirus
文献类型： 外文期刊
作者： Zhao, Haikuo 1 ; Zhou, Yong 2 ; Fan, Yuding 2 ; Jiang, Nan 2 ; Meng, Yan 2 ; Li, Yiqun 2 ; Xue, Mingyang 2 ; Xu, Chen 2 ; Guo, Weiliang 1 ; Liu, Wenzhi 2 ;
作者机构： 1.Hainan Univ, Coll Oceanog, Haikou, Peoples R China
2.Yangtze River Fisheries Res Inst, Chinese Acad Fishery Sci, Wuhan, Peoples R China
3.Hainan Univ, Coll Oceanog, Haikou 570228, Peoples R China
4.Yangtze River Fisheries Res Inst, Chinese Acad Fishery Sci, Wuhan 430223, Peoples R China
关键词： detection; droplet digital PCR (ddPCR); Oreochromis niloticus; real-time quantitative PCR (qPCR); tilapia parvovirus (TiPV)
期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.5； 五年影响因子：2.6 ）
ISSN： 0140-7775
年卷期： 2023 年 46 卷 3 期
页码：
收录情况： SCI
摘要： Tilapia parvovirus (TiPV) causes severe mortality rates in cultured tilapia, resulting in substantial losses to the fish industry. Droplet digital PCR (ddPCR) is a sensitive, accurate, and absolute quantitation method, plus it does not require a standard curve. Herein we report the development and application of a sensitive ddPCR-based method to rapidly detect and quantify TiPV. Optimal annealing temperature was determined to be 59.3 degrees C, and optimal primer and probe concentrations were 900 nmol/L and 250 nmol/L, respectively. Our ddPCR method was highly specific to TiPV and showed no cross-reactivity with other viruses. Further, the detection limit of ddPCR was 0.07 copies/mu l, being lower than that of real-time PCR (qPCR, 4.63 copies/mu l). We also investigated the ability of ddPCR to detect TiPV in 50 samples and compared the outcome with qPCR data in terms of sensitivity and accuracy. The results showed that the positive detection rate of ddPCR (32%) was higher than that of qPCR (18%). To conclude, our ddPCR method was effective at detecting TiPV in samples with low viral loads. We believe that its application can facilitate the surveillance of sources and transmission routes of TiPV.
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