实时RT-PCR法检测感染小鼠不同组织标本中的委内瑞拉马脑炎病毒  

Detection of Venezuelan equine encephalitis virus in different tissue samples of infected mice by real-time RT-PCR

作　　者：邓永强[1] 姜涛[1] 于曼[1] 陈水平[1] 秦成峰[1] 刘伯华[1] 祝庆余[1] 秦鄂德[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国人兽共患病学报》2006年第11期1035-1038,共4页Chinese Journal of Zoonoses

基　　金："十五"国家科技攻关计划项目(2003BA712A和2004BA519A32)

摘　　要：目的为委内瑞拉马脑炎病毒所致疾病的早期诊断和流行病学研究提供技术支持。方法建立了委内瑞拉马脑炎病毒特异、敏感和快速的实时RT-PCR法,并对委内瑞拉马脑炎病毒感染昆明小鼠的不同组织标本中的病毒载量进行了定量检测。结果本研究建立的实时RT-PCR法敏感(10TCID50/ml)、特异(对其它甲病毒成员,如东部马脑炎病毒和西部马脑炎病毒检测为阴性)。该方法可从委内瑞拉马脑炎病毒感染小鼠后第1d的血液、脾脏、脑组织中检测到不同含量的病毒核酸,而在感染后第3d仅从脾脏和脑组织中检测到病毒核酸。结论实时RT-PCR法不仅是一种敏感、准确的检测方法,可用来了解病毒感染量与疾病严重程度的关系和评价病毒病疫苗的免疫效果,而且也为VEEV疾病的临床诊断和流行病学监测提供工具。Venezuelan equine encephalitis virus （VEEV） is one of medically important human pathogen and potential biological warfare or bioterrorism agents. In order to develop a sensitive. specific and rapid technique of earlier surveillance and epidemiological research on VEEV disease, the real-time RT-PCR assay targeted capsid protein gene sequences of VEEV was developed and used to quantitatively detect the viral loads in different tissue samples of VEEV-infected mice. The results showed that real-time RT-PCR assay was sensitive （to detedct at least 10 TCID50/ml of viruses） and specific （not amplifying any other Alphaviruses, such as eastern equine encephalitis virus and western equine encephalitis virus）. In addition, virus RNA could be quantitatively detected from blood, kidney, liver, spleen and brain samples of VEEV-infected mice as early as 1 day post-inoculation, and from spleen and brain samples on the third day after inoculation of viruses, indicating that VEEV primarily replicates in spleen and brain of mouse. These suggest that real-time RT-PCR not only is a sensitive and accurate method to understand relationship between amount of virus infection-severe degree of diseases and assess the immunization efficacy of virus vaccine. but also have great potential in the clinical diagnosis and epidemiological surveillance of VEEV disease.

关 键 词：委内瑞拉马脑炎病毒 实时RT—PCR 

分 类 号：R373.1[医药卫生—病原生物学]