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
徐鸣明 1张英英 1展群岭 1何丰 1张亮 1金戈 1宋哲 1李亚军 2谢鹏1
作者信息
- 1. 重庆医科大学附属第一医院神经内科,重庆400016
- 2. 西安医学院附属医院神经内科,陕西西安710077
- 折叠
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.
Abstract
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.
关键词
博尔纳病/逆转录原位PCR/荧光定量PCR/感染Key words
borna disease virus/reverse transcription in situ PCR/real-time PCR/infection引用本文复制引用
基金项目
国家高技术研究发展计划(863计划)(2006AA02Z196)
出版年
2009
NETL
NSTL
维普
万方数据