title: Detection of Chlamydia pneumoniae in atherosclerotic tissue: a comparative study of PCR and immunocytochemistry.
abstract: The reported prevalence of Chlamydia pneumoniae in atherosclerotic tissue appears to depend on the detection system used. This introduces problems in determining the role of C. pneumoniae in atherosclerosis. This study analyses the sensitivity and performance of molecular diagnostic methods for the detection of C. pneumoniae and polymerase chain reaction (PCR) inhibitors in atheromatous tissue. Atherosclerotic tissue taken from 30 coronary endarterectomies, nine coronary arteries from explanted hearts, 16 carotid and two femoral endarterectomies are studied. Nested PCR (nPCR) assays targeting the PstI restriction fragment, the OmpA gene and the CRP operon of the chlamydial genome and immunocytochemistry (ICC) are used. Internal controls (IC) are constructed to co-amplify with the specific amplicons and identify the presence of inhibitor. The OmpA, PstI and CRP operon PCR assays had similar analytical sensitivities. However, the OmpA PCR was most affected by PCR inhibitors. Despite this, eight samples (14%) tested positive in the OmpA nPCR and no positives were found using the PstI or CRP operon nPCRs. Primary isolates of C. pneumoniae obtained from 12 patients with acute respiratory infection were positive in all three assays. Of the 48 specimens available for ICC, 33 (69%) were positive for chlamydial antigens. These included samples found positive by PCR. Dilution of samples to eliminate PCR inhibitors may have contributed to the discordant ICC and PCR results. The OmpA PCR, when used with an IC to identify samples with PCR inhibitors, is a reliable tool. However, the sensitivity of the ICC methods justifies their continued use.
authors.affiliation: Division of Cardiac and Vascular Sciences, St George's Hospital, University of London, Cranmer Terrace, UK.
authors.forename: G
authors.initials: G
authors.lastname: Arno
authors.forename: J C
authors.initials: JC
authors.lastname: Kaski
authors.forename: D A
authors.initials: DA
authors.lastname: Smith
authors.forename: J
authors.initials: J
authors.lastname: Akiyu
authors.forename: B
authors.initials: B
authors.lastname: Zal
authors.forename: B C
authors.initials: BC
authors.lastname: Millar
authors.forename: J E
authors.initials: JE
authors.lastname: Moore
authors.forename: C
authors.initials: C
authors.lastname: Baboonian
mesh_terms: D000328:Adult; D000368:Aged; D050197:Atherosclerosis / Q000382:microbiology*; D001431:Bacteriological Techniques / Q000379:methods; D002690:Chlamydia Infections / Q000175:diagnosis*; D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction