关键词: 对虾, 白斑综合征, 环介导等温扩增, 快速检测, shrimp, white spot syndrome virus, loop-mediated isothermal amplification, rapid detection

Abstract: In the present study, four sets of five specific primers were designed based on the conservative sequence of WSSV genome using PrimerExplorer V4. Five specific primers including one set of inner primers FIP and BIP, one set of outer primers F3 and B3, and one loop primer LF or LB. A visual WSSV-LAMP was developed in which calcein was applied to be detection indicator. Its detection limit for WSSV was determined by detecting the ten-fold diluted recombinant plasmid of pMD18-WSSV. The reaction temperature and time of WSSV-LAMP were at 63 ℃ for 60 min, respectively. Detection limit of the LAMP method for the WSSV DNA in which WSSV-2, WSSV-3 and WSSV-4 were determined to be 2.25 ~ 0.225 copies/μL, 2.25 ~ 0.225 copies/μL and 225 ~ 22.5 copies/μL, respectively. The LAMP assay could be finished within an hour, requiring only a regular laboratory water bath or heat block for reaction and the result could easily be detected using visual observation. Therefore, it is very suitable for detection in the fields that would provide a rapid detection approach for early WSSV infection in aquaculture.