双重PCR法检测痰标本对早期诊断军团菌肺炎的意义

首都医科大学学报 ›› 2001, Vol. 22 ›› Issue (4): 330-334.

双重PCR法检测痰标本对早期诊断军团菌肺炎的意义

张沪生¹, 金建敏¹, 陈东宁¹, 秦杰¹, 邱琼², 万超群²

1. 首都医科大学附属北京同仁医院呼吸内科;2. 中国预防医学科学院流行病学微生物学研究所

收稿日期:2000-09-05 修回日期:1900-01-01 出版日期:2001-10-15 发布日期:2001-10-15

Evaluation of Duplex PCR Method for Sputum Samples in Early Diagnosis of Legionella Pneumonia

Zhang Husheng¹, Jin Jianmin¹, Chen Dongning¹, Qin Jie¹, Qiu Qiong², Wan Chaoqun²

1. Department of Respiratory Disease, Beijing Tongren Hospital, Affiliate of Capital University of Medical Sciences;2. Insitute Epidemiology Microbiology, Chinese Academy of Preventiv

Received:2000-09-05 Revised:1900-01-01 Online:2001-10-15 Published:2001-10-15

摘要/Abstract

摘要： 为探讨双重PCR方法(DPCR)检测痰标本中军团菌DNA在早期诊断军团菌肺炎方面的意义,采用DPCR方法对军团菌肺炎组、临床疑似军团菌肺炎组、普通肺炎组和肺结核组4组患者留取的痰标本进行检测.DPCR过程采用根据军团菌16SrRNA基因和mip基因设计的2对引物,同时扩增军团菌386bp16SrRNA基因片段和206bpmip基因片段.结果:军团菌肺炎组的DPCR阳性检出率为100%,明显高于其他3组(分别为17.24%、0、0,P均<0.01).模拟痰标本DPCR的最低检出限为1×10³cfu/mL.初步研究表明,DPCR方法检测痰标本中的军团菌DNA具有较好的敏感性、特异性和稳定性,此法在临床早期诊断军团菌肺炎方面具有一定的价值.

关键词: 双重PCR, 军团菌, 军团菌肺炎, 痰标本

Abstract: To evaluate a Duplex PCR(DPCR)method in early diagnosis of legionella pneumonia by detecting legionella DNA in sputum samples, the sputum samples of patients from four groups, including legionella pneumonia group(n=15), clinical suspected legionella pneumonia group(n=29), ordinary pneumonia group(n=31)and pulmonary tuberculosis group(n=15)were detected with the DPCR method. Two different sets of oligonucleotide primer were stimultaneously used to amplify 386 bp 16SrRNA gene fragment and 206 bp mip gene fragment of legionella during the DPCR process. The DPCR positive rate of legionella pneumonia group(100%)was significantly higher than that of the other three groups(17.24%, 0, 0, respectively, P<0.01). The lowest detection level was 1×10³ cfu/mL for the simulated sputum sample. Primary study showed the DPCR method adopted for detecting legionella DNA in sputum samples had satisfactory sensitivity, specificity and stability. The method has its value in early diagnosis of legionella pneumonia.

Key words: duplex PCR, legionella pneumonia, sputum sample

中图分类号:

R517.9

张沪生;金建敏;陈东宁;秦杰;邱琼;万超群. 双重PCR法检测痰标本对早期诊断军团菌肺炎的意义[J]. 首都医科大学学报, 2001, 22(4): 330-334.

Zhang Husheng;Jin Jianmin;Chen Dongning;Qin Jie;Qiu Qiong;Wan Chaoqun. Evaluation of Duplex PCR Method for Sputum Samples in Early Diagnosis of Legionella Pneumonia[J]. Journal of Capital Medical University, 2001, 22(4): 330-334.

[1]	郭晶晶, 方茜, 刘亚楠, 曲沛, 马雪瀛, 王爽, 方伟, 周茹, 张响, 汤云霞, 杨思园, 于凤婷, 王雅杰. 31例新型冠状病毒感染病例咽拭子与痰标本病毒核酸检测的比较[J]. 首都医科大学学报, 2020, 41(3): 336-339.
[2]	金建敏;张沪生;陈东宁. 嗜肺军团菌与非嗜肺军团菌所致社区获得性肺炎的临床对比分析[J]. 首都医科大学学报, 2008, 29(3): 348-353.
[3]	苏文娟;陈秉良;关绍晨;党斌温;胡虹英;李小莹;涂国红. 院内和社区获得性军团菌肺炎71例临床分析[J]. 首都医科大学学报, 1999, 20(2): 113-115.