Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1975 Nov;28(11):910–914. doi: 10.1136/jcp.28.11.910

Laboratory procedures for the isolation of chlamydia trachomatis from the human genital tract.

P Reeve, J Owen, J D Oriel
PMCID: PMC475892  PMID: 1236632

Abstract

The technique of isolating Chlamydia trachomatis from the human gential tract by centrifuging clinical specimens on to cell monolayers with subsequent incubation has been improved and simplified. Gentamicin in the media was found to be superior to streptomycin in reducing bacterial contamination of specimens. The infectivity of chlamydial suspensions of laboratory cultured material was significantly reduced by storage at +4 degrees C for more than 48 hours, and by immediate freezing to -70 degrees C. When compared with immediate processing of the specimens, freezing to -70 degrees C was found to reduce the isolation rate of C.trachomatis from men with non-gonococcal urethritis (NGU) by approximately 20%. McCoy cells pretreated with idoxuridine were compared with irradiated McCoy cells for the isolation of C. trachomatis from clinical specimens. There was no significant difference in sensitivity between the two systems, but the former is considerably simpler. The effect of the centrifugal force used for inoculating specimens on to the cell monolayers on the isolation rate of C. trachomatis was studied in groups of men with NGU. Maximal isolation rates were obtained with forces of about 3000 G, which were not significantly raised by further increasing the force used. It is suggested that the isolation of C. trachomatis from the genital tract is now well within the capacity of any laboratory equipped with simple cell culture facilities.

Full text

PDF
912

Selected References

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

  1. Blyth W. A., Taverne J. Cultivation of TRIC agents: a comparison between the use of BHK-21 and irradiated McCoy cells. J Hyg (Lond) 1974 Feb;72(1):121–128. doi: 10.1017/s0022172400023287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Darougar S., Cubitt S., Jones B. R. Effect of high-speed centrifugation on the sensitivity of irradiated McCoy cell culture for the isolation of Chlamydia. Br J Vener Dis. 1974 Aug;50(4):308–312. doi: 10.1136/sti.50.4.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dunlop E. M., Freedman A., Garland J. A., Harper I. A., Jones B. R., Race J. W., Du Toit M. S., Treharne J. D. Infection by Bedsoniae and the possibility of spurious isolation. 2. Genital infection, disease of the eye, Reiter's disease. Am J Ophthalmol. 1967 May;63(5 Suppl):1073–1081. doi: 10.1016/0002-9394(67)94085-8. [DOI] [PubMed] [Google Scholar]
  4. Dunlop E. M., Vaughan-Jackson J. D., Darougar S., Jones B. R. Chlamydial infection. Incidence in 'non-specific' urethritis. Br J Vener Dis. 1972 Dec;48(6):425–428. doi: 10.1136/sti.48.6.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. FURNESS G., GRAHAM D. M., REEVE P. The titration of trachoma and inclusion blennorrhoea viruses in cell cultures. J Gen Microbiol. 1960 Dec;23:613–619. doi: 10.1099/00221287-23-3-613. [DOI] [PubMed] [Google Scholar]
  6. GORDON F. B., QUAN A. L. ISOLATION OF THE TRACHOMA AGENT IN CELL CULTURE. Proc Soc Exp Biol Med. 1965 Feb;118:354–359. doi: 10.3181/00379727-118-29841. [DOI] [PubMed] [Google Scholar]
  7. Gordon F. B., Dressler H. R., Quan A. L. Relative sensitivity of cell culture and yolk sac for detection of TRIC infection. Am J Ophthalmol. 1967 May;63(5 Suppl):1044–1048. doi: 10.1016/0002-9394(67)94080-9. [DOI] [PubMed] [Google Scholar]
  8. Hilton A. L., Richmond S. J., Milne J. D., Hindley F., Clarke S. K. Chlamydia A in the female genital tract. Br J Vener Dis. 1974 Feb;50(1):1–10. doi: 10.1136/sti.50.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hobson D., Johnson F. W., Rees E., Tait I. A. Simplified method for diagnosis of genital and ocular infections with Chlamydia. Lancet. 1974 Sep 7;2(7880):555–556. doi: 10.1016/s0140-6736(74)91879-0. [DOI] [PubMed] [Google Scholar]
  10. Kuo C., Wang S., Wentworth B. B., Grayston J. T. Primary isolation of TRIC organisms in HeLa 229 cells treated with DEAE-dextran. J Infect Dis. 1972 Jun;125(6):665–668. doi: 10.1093/infdis/125.6.665. [DOI] [PubMed] [Google Scholar]
  11. Oriel J. D., Powis P. A., Reeve P., Miller A., Nicol C. S. Chlamydial infections of the cervix. Br J Vener Dis. 1974 Feb;50(1):11–16. doi: 10.1136/sti.50.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Oriel J. D., Reeve P., Powis P., Miller A., Nicol C. S. Chlamydial infection. Isolation of Chlamydia from patients with non-specific genital infection. Br J Vener Dis. 1972 Dec;48(6):429–436. doi: 10.1136/sti.48.6.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Oriel J. D., Reeve P., Thomas B. J., Nicol C. S. Infection with Chlamydia group A in men with urethritis due to Neisseria gonorrhoeae. J Infect Dis. 1975 Apr;131(4):376–382. doi: 10.1093/infdis/131.4.376. [DOI] [PubMed] [Google Scholar]
  14. Reeve R., Taverne J. Strain differences in the behavior of TRIC agnets in cell cultures. Am J Ophthalmol. 1967 May;63(5 Suppl):1167–1173. doi: 10.1016/0002-9394(67)94099-8. [DOI] [PubMed] [Google Scholar]
  15. Richmond S. J., Hilton A. L., Clarke S. K. Chlamydial infection. Role of Chlamydia subgroup A in non-gonococcal and post-gonococcal urethritis. Br J Vener Dis. 1972 Dec;48(6):437–444. doi: 10.1136/sti.48.6.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wentworth B. B., Alexander E. R. Isolation of Chlamydia trachomatis by use of 5-iodo-2-deoxyuridine-treated cells. Appl Microbiol. 1974 May;27(5):912–916. doi: 10.1128/am.27.5.912-916.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wentworth B. B. Use of gentamicin in the isolation of subgroup A Chlamydia. Antimicrob Agents Chemother. 1973 Jun;3(6):698–702. doi: 10.1128/aac.3.6.698. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES