CAJ | 학술논문

目的建立脑心肌炎病毒（ EMCV） TaqMan real-time PCR检测方法。方法根据GenBank中公布的EMCV 3D基因保守区段设计并合成1对引物和1条TaqMan 探针，建立EMCV TaqMan real-time PCR检测方法，且对体系进行优化；对该法进行灵敏性、特异性验证；采用建立的方法对98份猪血清样本进行检测，并与ELISA结果进行比较。结果建立的EMCV TaqMan real-time PCR检测方法线性关系较好，以质粒标准品构建的标准曲线相关系数R2为0．995；灵敏性比普通PCR高100倍，且仅能特异性检出 EMCV；对猪血清样本的检测与 ELISA 法检测结果符合率为98．0％。结论已建立了EMCV TaqMan real-time PCR检测方法，该法灵敏性高、特异性好，可用于EMCV的检测及定量分析。
목적건립뇌심기염병독（ EMCV） TaqMan real-time PCR검측방법。방법근거GenBank중공포적EMCV 3D기인보수구단설계병합성1대인물화1조TaqMan 탐침，건립EMCV TaqMan real-time PCR검측방법，차대체계진행우화；대해법진행령민성、특이성험증；채용건립적방법대98빈저혈청양본진행검측，병여ELISA결과진행비교。결과건립적EMCV TaqMan real-time PCR검측방법선성관계교호，이질립표준품구건적표준곡선상관계수R2위0．995；령민성비보통PCR고100배，차부능특이성검출 EMCV；대저혈청양본적검측여 ELISA 법검측결과부합솔위98．0％。결론이건립료EMCV TaqMan real-time PCR검측방법，해법령민성고、특이성호，가용우EMCV적검측급정량분석。
Objective To establish a TaqMan real-time PCR assay for the detection of encephalo-myocarditis virus ( EMCV) .Methods Based on the conservative region of 3D gene of EMCV published in GenBank , a pair of primers and one TaqMan probe were designed and synthesized .Then a TaqMan real-time PCR assay was set up and the reactive system was optimized .The sensitivity and specificity of the assay was evaluated respectively .The TaqMan real-time PCR assay was then carried out to detect 98 randomly selected swine serum samples and the results were compared with those by using ELISA .Results The Ct value of the templates had a good linear relationship with the log starting quantity , with a correlation coefficient of 0.995.The TaqMan real-time PCR assay was only specific for EMCV and its sensitivity was 100 times higher than that of the ordinary PCR .The coincidence rate between the established assay and the ELISA assay was 98.0%in the detection of 98 blood samples.Conclusion The TaqMan real-time PCR assay for the detec-tion of EMCV was successfully established with advantages of high sensitivity and good specificity .It could be used for detection of EMCV and quantitative analysis .