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Development and comparative evaluation of real-time PCR and real-time RPA assays for detection of tilapia lake virus

文献类型： 外文期刊

作者： Wang, Yahui 1 ; Wang, Qing 1 ; Bergmann, Sven M. 3 ; Li, Yingying 1 ; Li, Bo 1 ; Lv, Yuefeng 1 ; Yin, Jiyuan 1 ; Yang, Guang 4 ; Qv, Yang 1 ; Wang, Yingying 1 ; Zeng, Weiwei 2 ;

作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Fishery Drug Dev, Key Lab Aquat Anim Immune Technol,Minist Agr, Guangzhou 510380, Guangdong, Peoples R China

2.Foshan Univ, Sch Life Sci & Engn, Guangdong Prov Key Lab Anim Mol Design & Precise, Foshan, Peoples R China

3.Friedrich Loeffler Inst FLI, Fed Res Inst Anim Hlth, Inst Infectol, Greifswald, Germany

4.Tianjin Agr Univ, Coll Fisheries, Tianjin, Peoples R China

5.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai, Peoples R China

关键词： Tilapia lake virus; Recombinase polymerase amplification; Real-time PCR; Detection

期刊名称：MOLECULAR AND CELLULAR PROBES （ 影响因子：3.285； 五年影响因子：2.781 ）

ISSN： 0890-8508

年卷期： 2021 年 60 卷

页码：

收录情况： SCI

摘要： Tilapia lake virus (TiLV) is a newly emerged pathogen responsible for high mortality and economic losses in the global tilapia industry. Early and accurate diagnosis is an important priority for TiLV disease control. In order to evaluate the methodology in the molecular diagnosis of TiLV, we compared newly developed quantitative realtime PCR (qPCR) and real-time recombinase polymerase amplification (real-time RPA) assays regarding their sensitivities, specificities and detection effect on clinical samples. Real-time RPA amplified the target pathogen in less than 30 min at 39 degrees C with a detection limit of 620 copies, while qPCR required about 60 min with a detection limit of 62 copies. Both assays were specific for TiLV and there were no cross-reactions observed with other common fish pathogens. The assays were validated using 35 tissue samples from clinically infected and 60 from artificially infected animals. The sensitivities for the real-time RPA and qPCR assays were 93.33 and 100%, respectively, and the specificity was 100% for both. Both methods have their advantages and can play their roles in different situations. The qPCR is more suitable for quantitative analysis and accurate detection of TiLV in a diagnostic laboratory, whereas real-time RPA is more suitable for the diagnosis of clinical diseases and preliminary screening for TiLV infection in poorly equipped laboratories as well as in fish farms.

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