扎伊尔型、苏丹型埃博拉病毒TaqMan探针实时定量PCR核酸检测方法的建立  

Detection of Zaire Ebola virus and Sudan Ebola virus by TaqMan real-time PCR assay

作　　者：贾佳[1] 康晓平[1] 李裕昌[1] 吴晓燕[1] 张雨[1] 张雪松[1] 霍耐凡 冉鑫[1] 杨银辉[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：《国际药学研究杂志》2015年第1期80-84,共5页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81171663);病原微生物生物安全国家重点实验室基金资助项目(SKLPBS1415);国家传染病重大专项基金资助项目(2011ZX10004-001;2013ZX1004802)

摘　　要：目的建立针对扎伊尔型(ZEBOV)及苏丹型埃博拉病毒(SEBOV)特异、灵敏和快速的Taq Man探针实时定量PCR核酸检测方法。方法选用世界卫生组织(WHO)参比实验室所使用的引物和探针序列,建立针对扎伊尔型及苏丹型埃博拉病毒的NP基因序列保守区的Taq Man探针实时定量PCR核酸检测方法,验证检测方法的灵敏性、特异性和稳定性,并与常规PCR检测方法进行比较。结果所建立的实时定量PCR方法可用于ZEBOV及SEBOV的特异性检测,较传统PCR方法反应快捷、灵敏,分别可达:ZEBOV 34拷贝/反应、SEBOV 24拷贝/反应;具有良好的特异性、稳定性和可重复性。结论建立了ZEBOV及SEBOV Taq Man探针实时定量PCR核酸检测方法,该方法可有望用于埃博拉病毒(EBOV)的实验室、临床及现场核酸检测。Objective To develop specific,sensitive and rapid Taq Man real-time quantification PCR assays for the detection of Zaire Ebola virus（ZEBOV）and Sudan Ebola virus（SEBOV）. Methods We selected primer and probe used by WHO reference laboratory,targeted the NP region of the viral genome,evaluated the specificity,sensitivity and reproducibility of each assay and compared it with original polymerase chain reation. Results The Zaire assay could detect 34 copies of the artificial plasmid which contains ZEBOV target sequence,while the Sudan assay could detect 24 copies of the artificial plasmid which contains SEBOV target sequence. In addition,the assay was highly specific and reduplicated for Ebola virus,and more rapid and sensitive than the original one. Conclusion We developed and evaluated Taq Man reverse transcription real time quantification PCR（RT-q PCR）assays for the rapid detection of ZEBOV and SEBOV. It seems that the assays are potentially suitable for diagnosis in the laboratories or field.

关 键 词：埃博拉病毒 扎伊尔型 埃博拉病毒 苏丹型 TaqMan探针实时定量PCR 核酸检测 

分 类 号：R440[医药卫生—诊断学]