逆转录原位PCR检测博尔纳病病毒感染

Reverse transcription in situ PCR detection of borna disease virus infection

目的:建立逆转录原位PCR检测博尔纳病病毒(BDV)的方法,并与其它检测方法进行比较.方法:构建包含BDV GFP-P24质粒的模拟BDV感染的OL细胞模型,检测其核酸的表达.对构建的OL细胞模型和各种阴性对照进行逆转录原位PCR的检测.使用逆转录原位PCR、荧光定量PCR和ELISA对BDV阳性的人和动物样本及阴性对照进行检测,并比较检测结果.结果:成功构建了含BDV-GFP-P24质粒模拟BDV感染的OL细胞.建立了逆转录原位PCR检测BDV的方法;对荧光定量PCR和ELISA的检测呈阳性的4个病例和检测呈阴性的10个病例,使用原位PCR检测可以得到基本一致的结果.结论:逆转录原位PCR原位可以用于检测人和动物的BDV感染和BDV感染机制研究.

AIM：To establish reverse transcription in situ PCR（RT ISPCR） detection of borna disease virus（BDV） infection and compare it with RT-PCR and ELISA detection.METHODS：The OL cells which used as BDV detection model were transfected by plasmid,BDV-GFP-P24,and tested by RT-PCR.All the BDV positive samples and negative controls（both human and animal） were tested by RT ISPCR,RT-PCR and ELISA.RESULTS：The BDV detection model was confirmed by RT-PCR.4 samples were detected positive and 10 samples were negative by RT IS PCR.The results were only slight different to that of RT-PCT（1 suspected positive when RT ISPCR is positive） and ELISA（1 suspected positive when RT ISPCR is negative）.CONCLUSION：RT ISPCR can be used to detection of human and animal infection of BDV and it can be used in the study of BDV infection mechanism.