Establishment of digital PCR method for detection of monkeypox virus
Objective To establish digital PCR detection method for monkeypox virus(MPXV)including universal type,West African branch and Central African branch,provide technical support for the precise prevention and control of MPXV at ports.Methods By comparing and analyzing the sequences of different branches of MPXV and other members of orthopoxvirus family,such as cowpox and vaccinia virus,3 sets of specific primers and probes were designed targeted MPXV G2R and C3L genes,and digital PCR methods were constructed to detect MPXV of universal type(G2R_GR),West African branch(G2R_WA)and Central African branch(C3L_CA),respectively.Results The established universal digital PCR detection method for MPXV could detect both West African branch and Central African branch,but could not detect other members of orthopoxvirus family such as cowpox and vccinia virus.Both the West African branch(G2R_WA)and Central African branch(C3L_CA)had good specifiaty.The detection sensitivity of the digital PCR method was higher,reached 45 copies/ml.Conclusion The digital PCR detection method constructed could achieve specific detection of MPXV of universal,West African branch and Central African branch,provided strong technical support for the prevention and control of MPXV at ports.
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