Establishment and application of a dual TaqMan real-time quantitative PCR method for lumpy skin disease virus and goatpox virus
In order to establish a method to rapidly identify lumpy skin disease virus(LSDV)and goatpox virus(GTPV),the whole gene sequences of LSDV and GTPV published in GenBank were compared and analyzed,and one pair of specific primers and two TaqMan probes were designed based on the conserved sequences of LSDV.After optimization of the reaction conditions,a dual TaqMan real-time quantitative PCR assay for LSDV and GTPV was established and evaluated for sensitivity,reproducibility,and specificity.The results showed that this method can successfully identify LSDV and GTPV from LSDV,GTPV,sheeppox virus,orf virus,bovine viral diarrhea virus and foot-and-mouth disease virus,which indicates good specificity.The lowest limit of detection for both LSDV and GTPV was 3.37 × 101 copies/μL.The coefficient of variation was less than 1％for both intra-and inter-group.The method was applied to 10 clinical samples,and the compliance rate was 100％.In conclusion,this method has good specificity,sensitivity and repro-ducibility,and can rapidly identify LSDV and GTPV in sample.It solves the current problem of overdiag-nosis of bovine lumpy skin disease,which is easily caused in China,and provides a new technical means for the comprehensive control of lumpy skin disease in China.
国家科技期刊平台
NSTL
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