Establishment of real-time fluorescent LAMP method for detection of Monkeypox virus

In order to establish a method for detecting Monkeypox virus（MPXV） with high specificity, specific primers were designed for OPG002 gene fragment in MPXV conserved region, and the volume of inner primers, loop primers and reaction temperature in loop-mediated isothermal amplification（LAMP） reaction system were optimized, and the specificity and sensitivity of the method was analyzed, and the method was applied to detect clinical samples and simulated samples. The results showed that the optimal reaction temperature and reaction time of the established MPXV real-time fluorescent LAMP method were 64 ℃ and 60 minutes, respectively. The method could detect MPXV specifically with a minimum detection limit of 25 copies/μL and a sensitivity 20 times higher than that of ordinary PCR. The results of 12 clinical samples tested by the method were negative, and the method could dectct simulated positive samples with plasmid concentration of 0.5×10~2～0.5×10~6 copies/μL. These results indicated that the MPXV real-time fluorescent LAMP method established in the study had the advantages of strong specificity, high sensitivity and simple operation, and could be used for rapid differential diagnosis of MPXV.