Feng Zizhao, Chu Xin, Han Minzhen, Yu Chenwei, Jiang Yousheng, Wang Hao, Lu Liqun, Xu Dan
College of Fisheries and life science, Shanghai Ocean University, Shanghai, China.
National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China.
J Fish Dis. 2022 Mar;45(3):461-469. doi: 10.1111/jfd.13575. Epub 2022 Jan 5.
Largemouth bass (Micropterus salmoides) is an important freshwater-cultured species in China. Recently, a lethal and epidemic disease caused by Micropterus salmoides rhabdovirus (MSRV) results in huge economic losses to the largemouth bass industry. Current diagnostics for detecting MSRV are limited in sensitivity and speed and are inconvenient to be used for non-laboratory detection. In this study, three rapid and convenient detection assays of MSRV by recombinase polymerase amplification (RPA) and lateral flow dipsticks (LFD), targeting the conserved sequences of the MSRV-SS N gene, are described. With these RPA methods, the detection could achieve within 50 min at 38°C. Both methods of RPA-AGE and RPA-LFD could detect the viral DNA as low as 170 copies/μl of the MSRV standard plasmid and were 100-fold more sensitive than that in the method of routine PCR. Meanwhile, these RPA methods were highly specific for the detection of MSRV and can be feasibly applied to the diagnostic of MSRV infection. In brief, RPA-AGE, RPA-LFD and RT-RPA-LFD provide convenient, rapid, sensitive and reliable methods that could improve field diagnosis of MSRV with limited machine resources, and would enhance the production of largemouth bass.
大口黑鲈（Micropterus salmoides）是中国重要的淡水养殖品种。近年来，由大口黑鲈弹状病毒（MSRV）引起的一种致死性流行病给大口黑鲈养殖业造成了巨大经济损失。目前用于检测MSRV的诊断方法在灵敏度和速度方面存在局限性，且不方便用于非实验室检测。在本研究中，描述了三种针对MSRV-SS N基因保守序列的、通过重组酶聚合酶扩增（RPA）和侧流试纸条（LFD）对MSRV进行快速便捷检测的方法。采用这些RPA方法，在38°C条件下50分钟内即可完成检测。RPA-AGE和RPA-LFD这两种方法均能检测低至170拷贝/μl的MSRV标准质粒病毒DNA，灵敏度比常规PCR方法高100倍。同时，这些RPA方法对MSRV检测具有高度特异性，可切实应用于MSRV感染的诊断。简而言之，RPA-AGE、RPA-LFD和RT-RPA-LFD提供了方便、快速、灵敏且可靠的方法，能够在机器资源有限的情况下改善MSRV的现场诊断，并将提高大口黑鲈的产量。
