Document Detail


Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens.
MedLine Citation:
PMID:  12005448     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
J Inman-Bamber; C Wan; T Gardam; R Vohra; A van Daal; P Timms
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Publication Detail:
Type:  Journal Article; Validation Studies    
Journal Detail:
Title:  Molecular and cellular probes     Volume:  16     ISSN:  0890-8508     ISO Abbreviation:  Mol. Cell. Probes     Publication Date:  2002 Feb 
Date Detail:
Created Date:  2002-05-13     Completed Date:  2002-09-10     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  8709751     Medline TA:  Mol Cell Probes     Country:  England    
Other Details:
Languages:  eng     Pagination:  57-61     Citation Subset:  IM    
Affiliation:
Cooperative Research Centre for Diagnostics, Queensland University of Technology, Brisbane, Australia.
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MeSH Terms
Descriptor/Qualifier:
Case-Control Studies
Chlamydophila Infections / diagnosis,  microbiology
Chlamydophila pneumoniae / isolation & purification*
Humans
Immunoenzyme Techniques / methods*
Polymerase Chain Reaction / methods*
Pulmonary Disease, Chronic Obstructive / microbiology*
Sensitivity and Specificity

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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