两种PCR方法检测狂犬病毒的敏感性和特异性比较

目的比较Nested-PCR和SYBR GreenⅠReal-Time PCR两种方法检测狂犬病毒的敏感性、特异性和时效性。方法对10倍连续稀释的狂犬病毒（疫苗株）核酸样品,采用Nested-PCR和SYBR GreenⅠReal-Time PCR方法进行平行检测,比较两种方法的敏感性;同时以登革热2型病毒、诺如病毒、星状病毒、EV71和乙型脑炎病毒核酸对两种方法的特异性进行评价。结果 SYBR GreenⅠReal-Time PCR方法可检测出2.3×106copies/μl核酸分子,与Nested-PCR方法相比,敏感度提高10倍。两种检测方法的特异性均好,与登革热2型病毒、诺如病毒、星状病毒、EV71和乙型脑炎病毒核酸均无交叉反应。SYBR GreenⅠReal-Time PCR方法检测花费3h,检测时间比Nested-PCR方法节省2h。结论与Nested-PCR相比,SYBR GreenⅠReal-Time PCR是相对高效的检测狂犬病毒的方法。

The sensitivity and specificity of two PCR assays for detecting rabies virus

Objective The aim of the study was to compare the sensitivity,specificity and time-consuming of Nested-PCR and SYBR GreenⅠReal-Time PCR assays for detecting rabies virus（RABV）.Methods A series of 10-fold dilutions of a RABV（vaccine strain） DNA was used for detection of the sensitivity of the Nested-PCR and SYBR GreenⅠReal-Time PCR assays.EV71,Dengue type 2 virus,Norovirus,Astrovirus and Japanese encephalitis virus（JEV） were used to test the specificity of the assays.Results Sensitivity of SYBR GreenⅠReal-Time PCR assay was 2.3×106 copies /μl which were 10 times more sensitive than that of Nested-PCR assay.There was no crossing-reaction with Dengue type 2 virus,Astrovirus,Norovirus,EV71 and JEV.The time-consuming of the SYBR GreenⅠreal-time PCR amplification was about 3 hours,2 hours shorter than Nested-PCR.Conclusion For detection of Rabies Virus,SYBR GreenⅠReal-Time PCR is a more sensitive and rapid assay compared with Nested-PCR.