An Economical Tandem Multiplex Real-Time PCR Technique for the Detection of a Comprehensive Range of Respiratory Pathogens

被引：50

作者：

Chidlow, Glenys R. ^{[1
,2
]}

Harnett, Gerry B. ^{[1
]}

Shellam, Geoffrey R. ^{[2
]}

Smith, David W. ^{[1
,2
]}

机构：

[1] Univ Western Australia, Queen Elizabeth II Med Ctr, Dept Microbiol, Pathwest Lab Med WA, Nedlands, WA 6009, Australia

[2] Univ Western Australia, Sch Biomed Biomol & Chem Sci M502, Nedlands, WA 6009, Australia

来源：

VIRUSES-BASEL | 2009年 / 1卷 / 01期

关键词：

respiratory pathogens; tandem multiplex real-time PCR; mixed infections; POLYMERASE-CHAIN-REACTION; REVERSE TRANSCRIPTION-PCR; COMMUNITY-ACQUIRED PNEUMONIA; TRACT INFECTIONS; SYNCYTIAL-VIRUS; NESTED-PCR; NASOPHARYNGEAL SAMPLES; ACUTE BRONCHIOLITIS; HUMAN RHINOVIRUSES; CLINICAL-SAMPLES;

D O I：

10.3390/v1010042

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

This study used real-time PCR assays to screen small sample volumes for a comprehensive range of 35 respiratory pathogens. Initial thermocycling was limited to 20 cycles to avoid competition for reagents, followed by a secondary real-time multiplex PCR. Supplementary semi-nested human metapneumovirus and picornavirus PCR assays were required to complete the acute respiratory pathogen profile. Potential pathogens were detected in 85 (70%) of pernasal aspirates collected from 121 children with acute respiratory symptoms. Multiple pathogens were detected in 29 (24%) of those samples. The tandem multiplex real-time PCR was an efficient method for the rapid detection of multiple pathogens.

引用

页码：42 / 56

页数：15

共 47 条

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