Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1992 Nov;30(11):2847–2851. doi: 10.1128/jcm.30.11.2847-2851.1992

Detection of Chlamydia trachomatis in endocervical specimens by polymerase chain reaction.

M J Loeffelholz 1, C A Lewinski 1, S R Silver 1, A P Purohit 1, S A Herman 1, D A Buonagurio 1, E A Dragon 1
PMCID: PMC270540  PMID: 1452654

Abstract

A rapid and sensitive polymerase chain reaction (PCR)-based assay for detection of Chlamydia trachomatis in cervical specimens is described. This assay consists of (i) sample preparation which avoids the use of heat, centrifugation, or organic extractions; (ii) rapid, two-temperature PCR amplification of C. trachomatis cryptic plasmid sequences; and (iii) capture and colorimetric detection of amplified DNA in microwell plates. PCR was compared with culture by using 503 cervical specimens. After resolution of discrepant specimens with a confirmatory PCR assay directed against the chlamydial major outer membrane protein gene, PCR had a sensitivity of 97% and a specificity of 99.7% while culture had a sensitivity of 85.7% and a specificity of 100%. In a separate study, PCR was compared with a direct specimen enzyme immunoassay (Chlamydiazyme; Abbott Diagnostics) by using 375 cervical specimens. After resolution of discrepant specimens, PCR had a sensitivity and specificity of 100%, while the enzyme immunoassay had a sensitivity of 58.8% and a specificity of 100%.

Full text

PDF
2847

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barnes R. C. Laboratory diagnosis of human chlamydial infections. Clin Microbiol Rev. 1989 Apr;2(2):119–136. doi: 10.1128/cmr.2.2.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bobo L., Coutlee F., Yolken R. H., Quinn T., Viscidi R. P. Diagnosis of Chlamydia trachomatis cervical infection by detection of amplified DNA with an enzyme immunoassay. J Clin Microbiol. 1990 Sep;28(9):1968–1973. doi: 10.1128/jcm.28.9.1968-1973.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Claas H. C., Wagenvoort J. H., Niesters H. G., Tio T. T., Van Rijsoort-Vos J. H., Quint W. G. Diagnostic value of the polymerase chain reaction for Chlamydia detection as determined in a follow-up study. J Clin Microbiol. 1991 Jan;29(1):42–45. doi: 10.1128/jcm.29.1.42-45.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Comanducci M., Ricci S., Cevenini R., Ratti G. Diversity of the Chlamydia trachomatis common plasmid in biovars with different pathogenicity. Plasmid. 1990 Mar;23(2):149–154. doi: 10.1016/0147-619x(90)90034-a. [DOI] [PubMed] [Google Scholar]
  5. Dutilh B., Bébéar C., Rodriguez P., Vekris A., Bonnet J., Garret M. Specific amplification of a DNA sequence common to all Chlamydia trachomatis serovars using the polymerase chain reaction. Res Microbiol. 1989 Jan;140(1):7–16. doi: 10.1016/0923-2508(89)90053-3. [DOI] [PubMed] [Google Scholar]
  6. Gratton C. A., Lim-Fong R., Prasad E., Kibsey P. C. Comparison of a DNA probe with culture for detecting Chlamydia trachomatis directly from genital specimens. Mol Cell Probes. 1990 Feb;4(1):25–31. doi: 10.1016/0890-8508(90)90036-y. [DOI] [PubMed] [Google Scholar]
  7. Hatt C., Ward M. E., Clarke I. N. Analysis of the entire nucleotide sequence of the cryptic plasmid of Chlamydia trachomatis serovar L1. Evidence for involvement in DNA replication. Nucleic Acids Res. 1988 May 11;16(9):4053–4067. doi: 10.1093/nar/16.9.4053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lefebvre J., Laperrière H., Rousseau H., Massé R. Comparison of three techniques for detection of Chlamydia trachomatis in endocervical specimens from asymptomatic women. J Clin Microbiol. 1988 Apr;26(4):726–731. doi: 10.1128/jcm.26.4.726-731.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ostergaard L., Birkelund S., Christiansen G. Use of polymerase chain reaction for detection of Chlamydia trachomatis. J Clin Microbiol. 1990 Jun;28(6):1254–1260. doi: 10.1128/jcm.28.6.1254-1260.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Peterson E. M., Markoff B. A., Schachter J., de la Maza L. M. The 7.5-kb plasmid present in Chlamydia trachomatis is not essential for the growth of this microorganism. Plasmid. 1990 Mar;23(2):144–148. doi: 10.1016/0147-619x(90)90033-9. [DOI] [PubMed] [Google Scholar]
  11. Ratti G., Moroni A., Cevenini R. Detection of Chlamydia trachomatis DNA in patients with non-gonococcal urethritis using the polymerase chain reaction. J Clin Pathol. 1991 Jul;44(7):564–568. doi: 10.1136/jcp.44.7.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Tung C. H., Rudolph M. J., Stein S. Preparation of oligonucleotide-peptide conjugates. Bioconjug Chem. 1991 Nov-Dec;2(6):464–465. doi: 10.1021/bc00012a015. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES