Huang Qian-Jun, Chen Yu, Liu Hong, St-Hilaire Sophie, Gao Shuai, MacKinnon Brett, Zhu Song-Qi, Wen Zhi-Qing, Jia Peng, Zheng Xiao-Cong
Department of Infectious Diseases and Public Health, Jockey Club College of Veterinary Medicine, City University of Hong Kong, Hong Kong.
University of Shanghai for Science and Technology, Shanghai, 200093, China.
J Virol Methods. 2022 Feb;300:114377. doi: 10.1016/j.jviromet.2021.114377. Epub 2021 Nov 23.
A rapid and simple real-time recombinase polymerase amplification (RPA) assay was developed to detect decapod iridescent virus 1 (DIV1). The assay was developed using optimized primers and probes designed from the conserved sequence of the DIV1 major capsid protein (MCP) gene. Using the optimized RPA assay, the DIV1 test was completed within 20 min at 39 ℃. The RPA assay was specific to DIV1 with a detection limit of 2.3 × 10 copies/reaction and there was no cross-reactivity with the other aquatic pathogens (WSSV, IHHNV, NHPB, VpAHPND, EHP, IMNV, YHV-1 and GAV) tested. Four out of 45 field-collected shrimp samples tested positive for DIV1 by real-time RPA. The same assay results were obtained by both methods. Thus, the real-time RPA assay developed could be a simple, rapid, sensitive, reliable and affordable method for the on-site diagnosis of DIV1 infection and has significant potential in helping to control DIV1 infections and reduce economic losses to the shrimp industry.
开发了一种快速简便的实时重组酶聚合酶扩增（RPA）检测方法来检测十足目虹彩病毒1（DIV1）。该检测方法是使用从DIV1主要衣壳蛋白（MCP）基因的保守序列设计的优化引物和探针开发的。使用优化的RPA检测方法，在39℃下20分钟内即可完成DIV1检测。该RPA检测方法对DIV1具有特异性，检测限为2.3×10拷贝/反应，并且与测试的其他水生病原体（WSSV、IHHNV、NHPB、VpAHPND、EHP、IMNV、YHV-1和GAV）没有交叉反应。在现场采集的45份虾样本中，有4份通过实时RPA检测出DIV1呈阳性。两种方法获得了相同的检测结果。因此，所开发的实时RPA检测方法可能是一种用于DIV1感染现场诊断的简单、快速、灵敏、可靠且经济实惠的方法，在帮助控制DIV1感染和减少对虾产业的经济损失方面具有巨大潜力。
