摘要
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.
出处
《第四军医大学学报》
北大核心
2009年第23期2862-2865,共4页
Journal of the Fourth Military Medical University
基金
国家高技术研究发展计划(2006AA02Z196)
关键词
博尔纳病
逆转录原位PCR
荧光定量PCR
感染
borna disease virus
reverse transcription in situ PCR
real-time PCR
infection