双重PCR方法早期诊断军团菌肺炎意义的初步探索  

Evaluation of a duplex PCR method in early diagnosis of Legionella pneumonia

作　　者：张沪生[1] 金建敏[1] 邱琼[2] 秦杰[1] 李松涛[1] 万超群[2] 

机构地区：[1]首都医科大学附属北京同仁医院呼吸内科,北京100730 [2]中国预防医学科学院流行病学微生物学研究所

出　　处：《中国人兽共患病杂志》2002年第1期82-88,共7页Chinese Journal of Zoonoses

基　　金：北京市卫生局科研基金项目 ( 98-0 1)

摘　　要：目的　探讨双重PCR方法 (DPCR)检测痰及支气管肺泡灌洗液 (BALF)中军团菌DNA在早期诊断军团菌肺炎方面的意义。方法　两对引物可扩增军团菌 386bp 16SrRNA基因片段和 2 0 6bpmip基因片段。采用DPCR方法对军团菌肺炎组 (15例 )、临床可疑军团菌肺炎组 (14例 )和普通肺炎组 (2 5例 )三组患者在病程早期留取的痰及BALF标本进行检测。结果　军团菌肺炎组的所有标本包括 2 5份痰、8份BALF的DPCR结果均为阳性 ,而普通肺炎组的所有标本包括 32份痰和 16份BALF均为DPCR阴性。临床可疑军团菌肺炎组留取的 2 0份痰和 8份BALF中 3例患者的 6份痰、2份BALF标本呈DPCR阳性。同一患者留取的多份标本呈现相同的DPCR结果。模拟标本的DPCR最低检出限为 1× 10 3 cfu/ml。结论　初步结果显示 :采用DPCR方法检测痰和BALF标本中的军团菌DNA具有较好的敏感性、特异性和稳定性 。Aim To evaluate a Duplex PCR (DPCR) method in early diagnosis of Legionella pneumonia by detecting the DNA in sputa and bronchoalvelar lavage fluids (BALF).Methods Two different sets of oligonucleotide primer were stimultaneously used to amplify 386bp 16SrRNA gene fragment and 206bp mip gene fragment of L.pneumonia.The sputa and BALF of patients from three groups,including L.pneumonia group (n=15),clinical suspected L.pneumonia group (n=14) and ordinary pneumonia group(n=25) were collected at early course of disease.All samples were detected with DPCR method for DNA.Results All the samples collected from L.pneumonia group,including 25 sputa and 8 BALF showed DPCR positive.The samples of ordinary pneumonia group including 32 sputa and 16 BALF were DPCR negative.Among the 20 sputa and 8 BALF of clinical suspected L.pneumonia group,6 sputa and 2 BALF from 3 patients showed positive DPCR results.The different samples of one patient showed the same DPCR results.The lowest detection level of simulated samples were 1×10 3 cfu/ml.Conclusions Primary study shows the DPCR method satisfactory sensitivity,specificity and stability.The method has its value in early diagnosis of L.pneumonia.

关 键 词：双重PCR 军团菌肺炎 早期诊断 

分 类 号：R517.9[医药卫生—内科学] R563.104[医药卫生—临床医学]