痘病毒科不同病毒属特异的PCR检测方法的建立

目的:建立可准确、快速地鉴别诊断可感染人的不同属痘病毒的特异PCR方法。方法:设计针对正痘病毒属、副痘病毒属和传染性软疣病毒属的多对特异引物,并制备相应的DNA模板,针对不同的模板优化引物与反应条件,分别进行检测筛选,建立病毒属特异的单独与多重PCR方法。结果:单一模板的PCR扩增反应中,正痘病毒的检测敏感性可达101拷贝/μL(引物为OPEaL-F1880/OPEaL-R2057),副痘病毒的检测敏感性可达101拷贝/μL(引物为PP2/PP3),传染性软疣病毒的检测敏感性为100 pg/μL体系(引物为MCV1/MCV2);混合模板的PCR扩增反应中,各属特异的引物均可获得预期大小的特异片段。结论:我们建立的PCR诊断方法,可用于痘病毒科不同病毒属感染的实验室特异快速鉴别诊断。

Development of Genus Specific PCR Method for Detection of Poxviridae

Objective： To develope an accurate and rapid genus specific PCR method for detection of Poxviridae.Methods： The multi-pair of primers and the corresponding DNA templates for Orthopoxvirus,Parapoxvirus and Molluscumcontagiosumvirus were prepared respectively,the different primers and reaction conditions were optimized for testing,and the genus specific single and multiple PCR methods were established.Results： In single template PCR amplification reaction,the detection sensitivity of Orthopoxvirus up to 101 copies / μL（primer OPEaL-F1880 / OPEaL-R2057）,Parapoxvirus detection sensitivity up to 101 copies / μL（primer PP2 / PP3）,and Molluscumcontagio-sumvirus sensitivity up to 100 pg / μL system（primer MCV1 / MCV2）.In multi-templates PCR amplification reaction,the expected specific fragments were obtained by different specific primers.Conclusion： The PCR methods estab-lished can be used for rapid laboratory genus specific identification of Poxviridae.