Angela van Daal
Angela van Daal
Associate Professor
Bond University
Receive Notification of New Content
Subscribe to new articlesNotify me of all new articles.
Receive customized notification.
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, Queensland University of Technology; and P Timms
DATE: February 2002
SOURCE: Molecular and Cellular Probes, Vol. 16, Iss. 1, pp. 57-61
ABOUT THIS DOCUMENT:
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.
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.