Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Feb;33(2):283–286. doi: 10.1128/jcm.33.2.283-286.1995

Solid-phase C1q-directed bacterial capture followed by PCR for detection of Chlamydia trachomatis in clinical specimens.

P Herbrink 1, H A van den Munckhof 1, H G Niesters 1, W H Goessens 1, E Stolz 1, W G Quint 1
PMCID: PMC227933  PMID: 7714179

Abstract

An antigen capture system based on the binding of bacteria to solid-phase immobilized complement C1q followed by PCR for detection of Chlamydia trachomatis in clinical samples was developed and clinically evaluated. Comparison of C1q-directed antigen capture PCR with cell culture and direct PCR on 71 consecutive clinical specimens revealed an identical sensitivity. In this group, all 11 cell culture-positive samples were positive by direct PCR and C1q-directed antigen capture PCR. In addition, two samples found negative by cell culture were found positive by both direct PCR and C1q-directed antigen capture PCR. To further assess the sensitivity of C1q-directed antigen capture PCR, 20 clinical samples with one to five inclusions in cell culture and 20 clinical samples with 6 to 20 inclusions in cell culture were tested. Results obtained showed sensitivities of 95 and 90% for clinical samples with 6 to 20 and 1 to 5 inclusions in cell culture, respectively. Using C1q-coated solid phases, C1q-binding Chlamydia particles can be concentrated from large volumes with concomitant removal of inhibitors of PCR, allowing the use of large volumes of clinical samples for clinical testing. Since C1q has been shown to bind to a range of gram-negative bacteria, the newly developed technique has utility for a broad range of bacteria.

Full Text

The Full Text of this article is available as a PDF (169.7 KB).

Selected References

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

  1. Betz S. J., Isliker H. Antibody-independent interactions between Escherichia coli J5 and human complement components. J Immunol. 1981 Nov;127(5):1748–1754. [PubMed] [Google Scholar]
  2. Claas H. C., Melchers W. J., de Bruijn I. H., de Graaf M., van Dijk W. C., Lindeman J., Quint W. G. Detection of Chlamydia trachomatis in clinical specimens by the polymerase chain reaction. Eur J Clin Microbiol Infect Dis. 1990 Dec;9(12):864–868. doi: 10.1007/BF01967500. [DOI] [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. Clas F., Golecki J. R., Loos M. Electron microscopic study showing antibody-independent binding of C1q, a subcomponent of the first component of complement, to serum-sensitive salmonellae. Infect Immun. 1984 Sep;45(3):795–797. doi: 10.1128/iai.45.3.795-797.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clas F., Loos M. Antibody-independent binding of the first component of complement (C1) and its subcomponent C1q to the S and R forms of Salmonella minnesota. Infect Immun. 1981 Mar;31(3):1138–1144. doi: 10.1128/iai.31.3.1138-1144.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clas F., Schmidt G., Loos M. The role of the classical pathway for the bactericidal effect of normal sera against gram-negative bacteria. Curr Top Microbiol Immunol. 1985;121:19–72. doi: 10.1007/978-3-642-45604-6_3. [DOI] [PubMed] [Google Scholar]
  7. Gerritsen M. J., Olyhoek T., Smits M. A., Bokhout B. A. Sample preparation method for polymerase chain reaction-based semiquantitative detection of Leptospira interrogans serovar hardjo subtype hardjobovis in bovine urine. J Clin Microbiol. 1991 Dec;29(12):2805–2808. doi: 10.1128/jcm.29.12.2805-2808.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grimprel E., Sanchez P. J., Wendel G. D., Burstain J. M., McCracken G. H., Jr, Radolf J. D., Norgard M. V. Use of polymerase chain reaction and rabbit infectivity testing to detect Treponema pallidum in amniotic fluid, fetal and neonatal sera, and cerebrospinal fluid. J Clin Microbiol. 1991 Aug;29(8):1711–1718. doi: 10.1128/jcm.29.8.1711-1718.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hall R. T., Strugnell T., Wu X., Devine D. V., Stiver H. G. Characterization of kinetics and target proteins for binding of human complement component C3 to the surface-exposed outer membrane of Chlamydia trachomatis serovar L2. Infect Immun. 1993 May;61(5):1829–1834. doi: 10.1128/iai.61.5.1829-1834.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hedrum A., Lundeberg J., Påhlson C., Uhlén M. Immunomagnetic recovery of Chlamydia trachomatis from urine with subsequent colorimetric DNA detection. PCR Methods Appl. 1992 Nov;2(2):167–171. doi: 10.1101/gr.2.2.167. [DOI] [PubMed] [Google Scholar]
  11. Horn J. E., Hammer M. L., Falkow S., Quinn T. C. Detection of Chlamydia trachomatis in tissue culture and cervical scrapings by in situ DNA hybridization. J Infect Dis. 1986 Jun;153(6):1155–1159. doi: 10.1093/infdis/153.6.1155. [DOI] [PubMed] [Google Scholar]
  12. Kluytmans J. A., Goessens W. H., Mouton J. W., van Rijsoort-Vos J. H., Niesters H. G., Quint W. G., Habbema L., Stolz E., Wagenvoort J. H. Evaluation of Clearview and Magic Lite tests, polymerase chain reaction, and cell culture for detection of Chlamydia trachomatis in urogenital specimens. J Clin Microbiol. 1993 Dec;31(12):3204–3210. doi: 10.1128/jcm.31.12.3204-3210.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  14. Liang T. J., Isselbacher K. J., Wands J. R. Rapid identification of low level hepatitis B-related viral genome in serum. J Clin Invest. 1989 Oct;84(4):1367–1371. doi: 10.1172/JCI114308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Loos M., Clas F. Antibody-independent killing of gram-negative bacteria via the classical pathway of complement. Immunol Lett. 1987 Feb;14(3):203–208. doi: 10.1016/0165-2478(87)90102-7. [DOI] [PubMed] [Google Scholar]
  16. Loos M., Wellek B., Thesen R., Opferkuch W. Antibody-independent interaction of the first component of complement with Gram-negative bacteria. Infect Immun. 1978 Oct;22(1):5–9. doi: 10.1128/iai.22.1.5-9.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meddens M. J., Quint W. G., van der Willigen H., Wagenvoort J. T., v Dijk W. C., Lindeman J., Herbrink P. Detection of Chlamydia trachomatis in culture and urogenital smears by in situ DNA hybridization using a biotinylated DNA probe. Mol Cell Probes. 1988 Dec;2(4):261–269. doi: 10.1016/0890-8508(88)90010-2. [DOI] [PubMed] [Google Scholar]
  18. Megran D. W., Stiver H. G., Bowie W. R. Complement activation and stimulation of chemotaxis by Chlamydia trachomatis. Infect Immun. 1985 Sep;49(3):670–673. doi: 10.1128/iai.49.3.670-673.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Melchers W. J., Schift R., Stolz E., Lindeman J., Quint W. G. Human papillomavirus detection in urine samples from male patients by the polymerase chain reaction. J Clin Microbiol. 1989 Aug;27(8):1711–1714. doi: 10.1128/jcm.27.8.1711-1714.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  21. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  22. Stamm W. E., Tam M., Koester M., Cles L. Detection of Chlamydia trachomatis inclusions in Mccoy cell cultures with fluorescein-conjugated monoclonal antibodies. J Clin Microbiol. 1983 Apr;17(4):666–668. doi: 10.1128/jcm.17.4.666-668.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Taylor-Robinson D., Thomas B. J., Osborn M. F. Evaluation of enzyme immunoassay (Chlamydiazyme) for detecting Chlamydia trachomatis in genital tract specimens. J Clin Pathol. 1987 Feb;40(2):194–199. doi: 10.1136/jcp.40.2.194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tenner A. J., Lesavre P. H., Cooper N. R. Purification and radiolabeling of human C1q. J Immunol. 1981 Aug;127(2):648–653. [PubMed] [Google Scholar]
  25. Tenner A. J., Ziccardi R. J., Cooper N. R. Antibody-independent C1 activation by E. coli. J Immunol. 1984 Aug;133(2):886–891. [PubMed] [Google Scholar]
  26. Thewessen E. A., Freundt I., van Rijsoort-Vos J. H., Stolz E., Michel M. F., Wagenvoort J. H. Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens. J Clin Microbiol. 1989 Jun;27(6):1399–1400. doi: 10.1128/jcm.27.6.1399-1400.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tjiam K. H., van Eijk R. V., van Heijst B. Y., Tideman G. J., van Joost T., Stolz E., Michel M. F. Evaluation of the direct fluorescent antibody test for diagnosis of chlamydial infections. Eur J Clin Microbiol. 1985 Dec;4(6):548–552. doi: 10.1007/BF02013392. [DOI] [PubMed] [Google Scholar]
  28. Widjojoatmodjo M. N., Fluit A. C., Torensma R., Verdonk G. P., Verhoef J. The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol. 1992 Dec;30(12):3195–3199. doi: 10.1128/jcm.30.12.3195-3199.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wilde J., Eiden J., Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol. 1990 Jun;28(6):1300–1307. doi: 10.1128/jcm.28.6.1300-1307.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zeldis J. B., Lee J. H., Mamish D., Finegold D. J., Sircar R., Ling Q., Knudsen P. J., Kuramoto I. K., Mimms L. T. Direct method for detecting small quantities of hepatitis B virus DNA in serum and plasma using the polymerase chain reaction. J Clin Invest. 1989 Nov;84(5):1503–1508. doi: 10.1172/JCI114326. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES