Establishment of a real-time Recombinase Polymerase Amplification (RPA) for the detection of decapod iridescent virus 1 (DIV1)

J Virol Methods. 2021 Nov 23:114377. doi: 10.1016/j.jviromet.2021.114377. Online ahead of print.ABSTRACTA 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 × 101 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.PMID:34826518 | DOI:10.1016/j.jviromet.2021.114377

Source: Journal of Virological Methods - November 26, 2021 Category: Virology Authors: Qian-Jun Huang Yu Chen Hong Liu Sophie St-Hilaire Shuai Gao Brett MacKinnon Song-Qi Zhu Zhi-Qing Wen Peng Jia Xiao-Cong Zheng Source Type: research