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通过定量实时聚合酶链反应检测肺炎患者呼吸道分泌物中的流感嗜血杆菌。

Detection of Haemophilus influenzae in respiratory secretions from pneumonia patients by quantitative real-time polymerase chain reaction.

作者信息

Abdeldaim Guma M K, Strålin Kristoffer, Kirsebom Leif A, Olcén Per, Blomberg Jonas, Herrmann Björn

机构信息

Department of Clinical Microbiology, Uppsala University Hospital, S-751 85 Uppsala, Sweden.

出版信息

Diagn Microbiol Infect Dis. 2009 Aug;64(4):366-73. doi: 10.1016/j.diagmicrobio.2009.03.030. Epub 2009 May 15.

Abstract

A quantitative real-time polymerase chain reaction (PCR) based on the omp P6 gene was developed to detect Haemophilus influenzae. Its specificity was determined by analysis of 29 strains of 11 different Haemophilus spp. and was compared with PCR assays having other target genes: rnpB, 16S rRNA, and bexA. The method was evaluated on nasopharyngeal aspirates from 166 adult patients with community-acquired pneumonia. When 10(4) DNA copies/mL was used as cutoff limit for the method, P6 PCR had a sensitivity of 97.5% and a specificity of 96.0% compared with the culture. Of 20 culture-negative but P6 PCR-positive cases, 18 were confirmed by fucK PCR as H. influenzae. Five (5.9%) of 84 nasopharyngeal aspirates from adult controls tested PCR positive. We conclude that the P6 real-time PCR is both sensitive and specific for identification of H. influenzae in respiratory secretions. Quantification facilitates discrimination between disease-causing H. influenzae strains and commensal colonization.

摘要

开发了一种基于omp P6基因的定量实时聚合酶链反应(PCR)来检测流感嗜血杆菌。通过分析11种不同嗜血杆菌属的29株菌株确定了其特异性,并与具有其他靶基因(rnpB、16S rRNA和bexA)的PCR检测方法进行了比较。该方法在166例社区获得性肺炎成年患者的鼻咽抽吸物上进行了评估。当将10(4)个DNA拷贝/毫升用作该方法的截断限,与培养法相比,P6 PCR的敏感性为97.5%,特异性为96.0%。在20例培养阴性但P6 PCR阳性的病例中,18例通过fucK PCR确认为流感嗜血杆菌。84例成年对照的鼻咽抽吸物中有5例(5.9%)PCR检测呈阳性。我们得出结论,P6实时PCR在呼吸道分泌物中鉴定流感嗜血杆菌方面既敏感又特异。定量有助于区分致病的流感嗜血杆菌菌株和共生定植。

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