Rapid detection of Singapore grouper iridovirus by a recombinase polymerase amplification combined with lateral flow dipstick
Singapore grouper iridovirus(SGIV),a novel species of Ranavirus,caused more than 90％mortality in larval and juvenile groupers.Up to now,there is still a lack of effective prevention and control strategies for SGIV.Therefore,it is essential to develop convenient diagnostic methods for filed detection of SGIV without special equipment.In this study,to establish a rapid,sensitive and visualized method for the detection of SGIV in clinical samples,specific primers and probes were designed by targeting the SGIV ORF014L,and a recombinase poly-merase amplification(RPA)technique combined with lateral flow dipstick(LFD)(RPA-LFD)was developed for the detection of SGIV.The results showed that the RPA reaction specifically detected target fragment of SGIV within 20 min at 40.1 ℃ with the lowest detection limit of 102 copies/μL.The RPA-LFD reaction at a constant temperature of 42 ℃ for 8 min was able to visualize the results on the test strips with the lowest detection limit of 101 copies/μL,and showed no cross-reaction with other common aquatic pathogens.The coincidence rate of posit-ive test of clinical samples was consistent between RPA-LFD and PCR methods.Both RPA and RPA-LFD could specifically detect SGIV with lower limit than conventional PCR assay.Taken together,RPA-LFD assay developed in the present study provides a convenient,specific,sensitive and visualized method for on-site rapid detection of SGIV without special equipment.
万方数据
维普
