引用本文:姜丽娟,吴文娟,吴 海,周 键,吴 伟,李 涛,郭 建,王洪海,卢水华,李 瑶.利用荧光定量RT-PCR和PCR方法检测结核分枝杆菌复合群[J].中国临床新医学,2012,5(4):285-289.
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利用荧光定量RT-PCR和PCR方法检测结核分枝杆菌复合群
姜丽娟,吴文娟,吴 海,周 键,吴 伟,李 涛,郭 建,王洪海,卢水华,李 瑶
200433 上海,复旦大学生命科学学院遗传学研究所国家遗传工程重点实验室(姜丽娟,吴 海,吴 伟,王洪海,李 瑶);201508 上海,复旦大学附属公共卫生临床中心(吴文娟,周 键,李 涛,郭 建,卢水华)
摘要:
[摘要] 目的 建立一种新的检测结核分枝杆菌复合群的荧光定量试验方法(R/P分析),探讨R/P分析检测临床标本中结核分枝杆菌复合群的应用价值。方法 根据结核分枝杆菌复合群基因保守序列设计引物和探针构建质粒标准品。运用R/P分析检测54例确诊结核病人临床样本,检测的结果同时与培养法、荧光定量PCR法进行比较。 结果 不同临床标本用R/P分析诊断结核病,其敏感性高于荧光定量PCR法和培养法,阳性检出率分别为96.3%、83.3%和55.6%。结论 R/P方法是一种快速、特异的直接检测方法,它可以区分结核分枝杆菌复合群的死菌与活菌,因此可以更好的指导医生进行治疗,更准确地做出公共卫生决策。
关键词:  结核分枝杆菌复合群  TaqMan荧光定量PCR  快速检测
DOI:10.3969/j.issn.1674-3806.2012.04.02
分类号:R 52
基金项目:国家科技重大专项课题资助项目(编号:2009ZX10004-313)
Rapid detection of Mycobacterium tuberculosis complex by combining real-time RT-PCR and real-time PCR assay
JIANG Li-juan, WU Wen-juan, WU Hai, et al.
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China
Abstract:
[Abstract] Objective To develop a new real-time assay(R/P assay) for detection of Mycobacterium tuberculosis complex and evaluate the application value of R/P assay in detection of Mycobacterium tuberculosis complex (MTBC) in clinical specimens.Methods The primers and probe were designed and constituted standard sample. The R/P assay was applied to 54 specimens of the clinical tuberculosis cases and the results were compared with those of culture and real-time PCR.Results The positive rate of R/P assay,Real time PCR,culture by the different clinical specimens were 96.3%,83.3% and 55.6% respectively.Conclusion The R/P assay is a rapid and specific method for direct detection of MTBC, which also can distinguish viable and nonviable MTBC, thus may guide patient therapy, and public health decisions.
Key words:  Mycobacterium tuberculosis complex  TaqMan Real time PCR  Rapid detection