鼠类携带淋巴细胞脉络丛脑膜炎病毒实时荧光定量RT-PCR检测方法的建立  

Establishment of a real-time quantitative PCR technique to detect lymphocytic choriomeningitis virus carried by rodents

作　　者：胡群[1] 邹春颖[1] 马思杰[1] 童淑梅[1] 

机构地区：[1]大榭出入境检验检疫局,浙江宁波315812

出　　处：《中国病原生物学杂志》2014年第10期877-879,共3页Journal of Pathogen Biology

基　　金：宁波市自然科学基金项目(No.2013A610240)

摘　　要：目的建立检测鼠类携带淋巴细胞脉络丛脑膜炎病毒(lymphocytic choriomeningitis virus,LCMV)的实时荧光定量RT-PCR方法。方法根据LCMV核蛋白编码基因序列设计合成特异性引物对和TaqMan荧光探针,经优化反应体系和条件,建立LCMV实时荧光定量RT-PCR检测方法,然后进行灵敏度、特异性和重复性试验,并对79份宁波口岸捕获的鼠样品进行检测。结果建立实时荧光定量RT-PCR方法对鼠肺总RNA检测的灵敏度为20pg,是常规PCR方法的100倍;试验的重复性良好,CV值为0.85%;试验的特异性为100%。用该方法检测79份鼠样品有3份LCMV阳性,与常规RT-PCR结果一致。结论成功建立了鼠LCMV实时荧光定量RT-PCR检测方法,对于监测和防控鼠传LCM有重要意义。Objective To establish a real-time quantitative PCR technique to detect lymphocytic choriomeningitis virus（LCMV）carried by rodents. Methods In accordance with the nucleocapsid protein gene sequence of LCMV,apair of specific primers and a TaqMan fluorescent probe were designed and a real-time quantitative PCR assay for detection of LCMV was established.The technique＇s sensitivity,specificity,and reproducibility were evaluated,and this technique was used to test 79 samples from rodents caught at the Port of Ningbo. Results The detection limit for real-time quantitative PCR detection of total RNA extracted from rodent lungs was 20 pg,which was 100 times greater than that of routine RT-PCR.The technique in question had good reproducibility with a coefficient of variation of 0.85%.The technique had a specificity of 100%.Three of the 79 rodent samples were positive for LCMV,which matched the results of routine RTPCR. Conclusion A real-time quantitative PCR technique to detect LCMV carried by rodents was successfully established.This technique is extremely important to the monitoring and control of LCMV carried by rodents.

关 键 词：淋巴细胞脉络丛脑膜炎病毒 实时荧光定量RT-PCR 检测 鼠类 

分 类 号：R37[医药卫生—病原生物学]