中国兽医科学2024，Vol.54Issue(9)：1188-1194,7.DOI:10.16656/j.issn.1673-4696.2024.0157
牛结节性皮肤病病毒与山羊痘病毒双重TaqMan实时荧光定量PCR方法的建立
Establishment and application of a dual TaqMan real-time quantitative PCR method for lumpy skin disease virus and goatpox virus
摘要
Abstract
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.关键词
牛结节性皮肤病病毒/山羊疽病毒/双重TaqMan实时定量PCRKey words
lumpy skin disease virus/goatpox virus/dual TaqMan real-time quantitative PCR分类
农业科技引用本文复制引用
余翰维,景志忠,陈轶霞,冯秋媛,苏洋,张慧,邵晓龙,杨云凤,何小兵,陈国华,房永祥..牛结节性皮肤病病毒与山羊痘病毒双重TaqMan实时荧光定量PCR方法的建立[J].中国兽医科学,2024,54(9):1188-1194,7.基金项目
国家自然科学基金项目(32260874) （32260874）
国家动物疫病监测与流行病学调查计划项目(125161031) （125161031）
家畜疫病病原生物学国家重点实验室(应用)基础重大成果培育项目(SKLVEB2021CGQD03) （应用）
甘肃省科技计划项目(22JR5RA780) （22JR5RA780）
