A droplet digital PCR assay to detect Chinese rice-field eels rhabdovirus

文献类型： 外文期刊

第一作者： Liu, Wenzhi

作者： Liu, Wenzhi;Zhou, Yong;Jiang, Nan;Xu, Chen;Fan, Yuding;Zhong, Qiwang

作者机构：

关键词： Chinese rice-field eels (Monopterus albus); Chinese rice-field eels rhabdovirus; droplet digital PCR; real-time quantitative PCR

期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.2； 五年影响因子：2.2 ）

ISSN： 0140-7775

年卷期： 2024 年 47 卷 12 期

页码：

收录情况： SCI

摘要： Chinese rice-field eels rhabdovirus (CrERV) causes haemorrhagic disease in Chinese rice-field eels (Monopterus albus), leading to significant mortality and economic losses. Sensitive detection of CrERV nucleic acids is essential to control the spread of this pathogen and to treat infected individuals. Herein, we developed an efficient and sensitive droplet digital PCR (ddPCR) method to rapidly detect and quantify CrERV. The ddPCR assay optimal conditions were an annealing temperature of 53 degrees C, and primer and probe concentrations of 0.5 and 0.25 mu M, respectively. The assay had a diagnostic sensitivity of 0.23 copies/mu L, and was highly specific, showing no cross reactivity with other viruses (infectious haematopoietic necrosis virus, grass carp reovirus, spring viremia of carp virus, largemouth bass ranavirus, carp edema virus, Chinese giant salamander iridovirus, and white spot syndrome virus). Real-time quantitative PCR testing of 30 Chinese rice-field eels samples detected CrERV in 7 samples (23.3%), whereas ddPCR detected CrERV in 12 samples (40%), demonstrating its higher sensitivity. Thus, ddPCR represents an advanced method to absolutely quantify CrERV in infected fish with low virus concentrations, providing a valuable tool to manage the spread and impact of CrERV.

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