This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Maraha, B. Articles by van der Zee, A. Search for Related Content PubMed PubMed Citation Articles by Maraha, B. Articles by van der Zee, A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2004, p. 3937-3941, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.3937-3941.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Is the Perceived Association between Chlamydia pneumoniae and Vascular Diseases Biased by Methodology?

Regional Laboratory of Medical Microbiology, Dordrecht-Gorinchem,¹ Department of Clinical Microbiology,² Department of Surgery, St. Elisabeth Hospital,⁴ Department of Surgery, TweeSteden Hospital, Tilburg,³ Department of Clinical Microbiology, Amphia Hospital, Breda, The Netherlands⁵

Received 7 December 2003/ Returned for modification 26 January 2004/ Accepted 28 March 2004

Inter- and intralaboratory inconsistencies in detection rates of Chlamydia pneumoniae in vascular specimens have been demonstrated. In this study, 66 vascular tissue specimens from 66 patients with vascular disease were tested by three PCR assays: a 16S PCR-based reverse line blot (RLB) assay, a single-step PCR, and a nested PCR. Also, we explored the impacts of different DNA polymerase enzymes on the results based on gel electrophoresis and hybridization. The PCR results by gel electrophoresis in the single-step PCR depended on which DNA polymerase was used. All samples were negative with AmpliTaq Gold DNA polymerase, and 54.5% (36 of 66) were positive with the conventional Taq DNA polymerase. All samples were negative after hybridization with a C. pneumoniae-specific probe. In the nested PCR, all specimens were negative by gel electrophoresis and after hybridization. The RLB assay failed to detect C. pneumoniae in any specimen; however, 20 specimens were Chlamydia sp. positive. The sequence analysis of six of these samples demonstrated Chlamydia-like organisms. RLB detected Chlamydia sp. DNA in water and in the elution buffer after passage of the Qiagen columns (11 of 40). This study identified factors that may influence the detection of C. pneumoniae DNA in vascular tissues and consequently bias the perception of a link between C. pneumoniae and vascular diseases. The following are strongly recommended: to use DNA polymerases that have to be activated, to decontaminate with dUTP-uracil-DNA glycosylase, to hybridize with specific probes, to include sufficient controls, and to use molecular grade water.

This article has been cited by other articles:

• Diederen, B. M. W., van der Valk, P. D. L. P. M., Kluytmans, J. A. W. J., Peeters, M. F., Hendrix, R. (2007). The role of atypical respiratory pathogens in exacerbations of chronic obstructive pulmonary disease. Eur Respir J 30: 240-244 [Abstract] [Full Text]  
• Corsaro, D., Greub, G. (2006). Pathogenic Potential of Novel Chlamydiae and Diagnostic Approaches to Infections Due to These Obligate Intracellular Bacteria. Clin. Microbiol. Rev. 19: 283-297 [Abstract] [Full Text]  
• Maggi, R. G., Breitschwerdt, E. B. (2005). Potential Limitations of the 16S-23S rRNA Intergenic Region for Molecular Detection of Bartonella Species. J. Clin. Microbiol. 43: 1171-1176 [Abstract] [Full Text]  
• Ieven, M. M., Hoymans, V. Y. (2005). Involvement of Chlamydia pneumoniae in Atherosclerosis: More Evidence for Lack of Evidence. J. Clin. Microbiol. 43: 19-24 [Full Text]