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Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens
Molecular and Cellular Probes (2002)
  • J Inman-Bamber
  • C Wan
  • T Gardam
  • R Vohra
  • A van Daal, Queensland University of Technology
  • P Timms
Abstract
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.
Disciplines
Publication Date
February, 2002
Publisher Statement

Interim status: Citation only.

Inman-Bamber J., Wan, C., Gardham,T., Vohra, R., van Daal, A. and Timms, P. (2002) Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens. Molecular and Cellular Probes, Vol. 16, Iss. 1, pp. 57-61.

Copyright ©Elsevier B.V. 2002. All rights reserved.

Citation Information
J Inman-Bamber, C Wan, T Gardam, R Vohra, et al.. "Novel PCR-EIA method for the detection of Chlamydia pneumoniae in respiratory specimens" Molecular and Cellular Probes Vol. 16 Iss. 1 (2002)
Available at: http://works.bepress.com/angela_v_daal/15/