Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens

J Inman-Bamber; C Wan; T Gardam; R Vohra; A Van Daal; Peter Timms

doi:10.1006/mcpr.2001.0399

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Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens

Journal article

Peer reviewed

Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens

J Inman-Bamber, C Wan, T Gardam, R Vohra, A Van Daal and Peter Timms

Molecular and Cellular Probes, Vol.16(1), pp.57-61

2002

DOI: https://doi.org/10.1006/mcpr.2001.0399

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Abstract

PCR-enzyme immunoassay

chronic obstructive airways disease

chlamydia pneumoniae

We report the development of a microtitre plate-based PCR-EIA assay (ELAHA; Enzyme Linked Amplification and Hybridization Assay) for the sensitive and specific detection of Chlamydia pneumoniae in sputum samples from patients with chronic obstructive airways disease (COAD). Following PCR amplification of a segment of the chlamydial heat shock 60 protein gene, the 587 bp sized amplicon is captured onto the streptavidin coated surface of a microtitre plate using a C. pneumoniae specific biotinylated probe and the level of captured product is subsequently determined via a colorimetric reaction using an automated plate reader. The ELAHA is a simple, rapid and inexpensive method for detection of low levels of infectious agents and is readily adaptable to current clinical laboratory equipment. The assay was evaluated with a cohort of hospital respiratory patients: (i) COAD patients with acute exacerbation, (ii) COAD patients without exacerbation (stable) and (iii) a non-respiratory control group. The ELAHA produced 6/12 (50%) C. pneumoniae positives in the COAD with exacerbation group, 3/13 (23%) positives in the COAD without exacerbation group and only 1/6 (17%) positives in the control non-respiratory group. This sensitive and robust PCR-EIA method can provide clinically relevant diagnostic evidence of current C. pneumoniae infection contributing to serious respiratory tract diseases such as COAD. © 2002 Elsevier Science Ltd.

Details

Title: Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens
Authors: J Inman-Bamber (Author) - Queensland University of Technology
C Wan (Author) - Queensland University of Technology
T Gardam (Author) - Queensland University of Technology
R Vohra (Author) - Queensland University of Technology
A Van Daal (Author) - Queensland University of Technology
Peter Timms (Author) - Queensland University of Technology
Publication details: Molecular and Cellular Probes, Vol.16(1), pp.57-61
Publisher: Academic Press
Date published: 2002
DOI: 10.1006/mcpr.2001.0399
ISSN: 0890-8508
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99449855302621
Output Type: Journal article

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