Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 May;60(5):2040–2047. doi: 10.1128/iai.60.5.2040-2047.1992

Demonstration of chlamydial RNA and DNA during a culture-negative state.

S M Holland 1, A P Hudson 1, L Bobo 1, J A Whittum-Hudson 1, R P Viscidi 1, T C Quinn 1, H R Taylor 1
PMCID: PMC257113  PMID: 1373404

Abstract

Trachoma is a common blinding disease of humans caused by ocular infections with Chlamydia trachomatis. The cynomolgus monkey is a valuable primate model for the detection, pathobiology, and treatment of this infection. We have used this model system to compare the relative ability of tissue culture, direct fluorescence cytology, a modified polymerase chain reaction, and RNA blotting to detect C. trachomatis following primary infection and reinfection over 34 weeks. Six cynomolgus monkeys were given a primary ocular chlamydia infection, and 20 weeks later they were reinoculated with the same organism. All animals showed brisk inflammatory responses to the primary infection and milder inflammatory reactions to reinfection. All four diagnostic techniques detected chlamydia at 1 week after primary infection, but both nucleic acid detection methods suggested that organisms were present longer after primary infection than did either tissue culture or direct fluorescence cytology (16 weeks for RNA blotting versus 12 weeks for tissue culture). Following reinoculation at 20 weeks, the period of C. trachomatis detection by tissue culture or direct fluorescence cytology (4 weeks) was much shorter than after primary infection. In contrast, nucleic acid detection was positive for up to 5 weeks longer than tissue culture or direct fluorescence cytology. Both polymerase chain reaction and RNA blotting, which involved no amplification step, indicated the presence of organisms during the culture-negative period. These data suggest that live chlamydiae may remain at a site of infection and produce inflammation beyond the time at which standard microbiological techniques are able to detect them.

Full text

PDF
2040

Selected References

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

  1. 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]
  2. Bobo L., Munoz B., Viscidi R., Quinn T., Mkocha H., West S. Diagnosis of Chlamydia trachomatis eye infection in Tanzania by polymerase chain reaction/enzyme immunoassay. Lancet. 1991 Oct 5;338(8771):847–850. doi: 10.1016/0140-6736(91)91502-l. [DOI] [PubMed] [Google Scholar]
  3. Boguslawski S. J., Smith D. E., Michalak M. A., Mickelson K. E., Yehle C. O., Patterson W. L., Carrico R. J. Characterization of monoclonal antibody to DNA.RNA and its application to immunodetection of hybrids. J Immunol Methods. 1986 May 1;89(1):123–130. doi: 10.1016/0022-1759(86)90040-2. [DOI] [PubMed] [Google Scholar]
  4. Byrne G. I., Lehmann L. K., Landry G. J. Induction of tryptophan catabolism is the mechanism for gamma-interferon-mediated inhibition of intracellular Chlamydia psittaci replication in T24 cells. Infect Immun. 1986 Aug;53(2):347–351. doi: 10.1128/iai.53.2.347-351.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheema M. A., Schumacher H. R., Hudson A. P. RNA-directed molecular hybridization screening: evidence for inapparent chlamydial infection. Am J Med Sci. 1991 Nov;302(5):261–268. doi: 10.1097/00000441-199111000-00001. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Grayston J. T., Wang S. P., Yeh L. J., Kuo C. C. Importance of reinfection in the pathogenesis of trachoma. Rev Infect Dis. 1985 Nov-Dec;7(6):717–725. doi: 10.1093/clinids/7.6.717. [DOI] [PubMed] [Google Scholar]
  8. Griffais R., Thibon M. Detection of Chlamydia trachomatis by the polymerase chain reaction. Res Microbiol. 1989 Feb;140(2):139–141. doi: 10.1016/0923-2508(89)90047-8. [DOI] [PubMed] [Google Scholar]
  9. Holland S. M., Gaydos C. A., Quinn T. C. Detection and differentiation of Chlamydia trachomatis, Chlamydia psittaci, and Chlamydia pneumoniae by DNA amplification. J Infect Dis. 1990 Oct;162(4):984–987. doi: 10.1093/infdis/162.4.984. [DOI] [PubMed] [Google Scholar]
  10. Morrison R. P., Belland R. J., Lyng K., Caldwell H. D. Chlamydial disease pathogenesis. The 57-kD chlamydial hypersensitivity antigen is a stress response protein. J Exp Med. 1989 Oct 1;170(4):1271–1283. doi: 10.1084/jem.170.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Morrison R. P., Lyng K., Caldwell H. D. Chlamydial disease pathogenesis. Ocular hypersensitivity elicited by a genus-specific 57-kD protein. J Exp Med. 1989 Mar 1;169(3):663–675. doi: 10.1084/jem.169.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Pollard D. R., Tyler S. D., Ng C. W., Rozee K. R. A polymerase chain reaction (PCR) protocol for the specific detection of Chlamydia spp. Mol Cell Probes. 1989 Dec;3(4):383–389. doi: 10.1016/0890-8508(89)90017-0. [DOI] [PubMed] [Google Scholar]
  14. Rothermel C. D., Schachter J., Lavrich P., Lipsitz E. C., Francus T. Chlamydia trachomatis-induced production of interleukin-1 by human monocytes. Infect Immun. 1989 Sep;57(9):2705–2711. doi: 10.1128/iai.57.9.2705-2711.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schachter J., Caldwell H. D. Chlamydiae. Annu Rev Microbiol. 1980;34:285–309. doi: 10.1146/annurev.mi.34.100180.001441. [DOI] [PubMed] [Google Scholar]
  16. Stamm W. E. Diagnosis of Chlamydia trachomatis genitourinary infections. Ann Intern Med. 1988 May;108(5):710–717. doi: 10.7326/0003-4819-108-5-710. [DOI] [PubMed] [Google Scholar]
  17. Stephens R. S., Sanchez-Pescador R., Wagar E. A., Inouye C., Urdea M. S. Diversity of Chlamydia trachomatis major outer membrane protein genes. J Bacteriol. 1987 Sep;169(9):3879–3885. doi: 10.1128/jb.169.9.3879-3885.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Taylor H. R. Development of immunity to ocular chlamydial infection. Am J Trop Med Hyg. 1990 Apr;42(4):358–364. doi: 10.4269/ajtmh.1990.42.358. [DOI] [PubMed] [Google Scholar]
  19. Taylor H. R., Johnson S. L., Prendergast R. A., Schachter J., Dawson C. R., Silverstein A. M. An animal model of trachoma II. The importance of repeated reinfection. Invest Ophthalmol Vis Sci. 1982 Oct;23(4):507–515. [PubMed] [Google Scholar]
  20. Taylor H. R., Johnson S. L., Schachter J., Caldwell H. D., Prendergast R. A. Pathogenesis of trachoma: the stimulus for inflammation. J Immunol. 1987 May 1;138(9):3023–3027. [PubMed] [Google Scholar]
  21. Taylor H. R., Maclean I. W., Brunham R. C., Pal S., Whittum-Hudson J. Chlamydial heat shock proteins and trachoma. Infect Immun. 1990 Sep;58(9):3061–3063. doi: 10.1128/iai.58.9.3061-3063.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Taylor H. R., Prendergast R. A., Dawson C. R., Schachter J., Silverstein A. M. An animal model for cicatrizing trachoma. Invest Ophthalmol Vis Sci. 1981 Sep;21(3):422–433. [PubMed] [Google Scholar]
  23. Taylor H. R., Rapoza P. A., West S., Johnson S., Munoz B., Katala S., Mmbaga B. B. The epidemiology of infection in trachoma. Invest Ophthalmol Vis Sci. 1989 Aug;30(8):1823–1833. [PubMed] [Google Scholar]
  24. Taylor H. R., Siler J. A., Mkocha H. A., Muñoz B., Velez V., Dejong L., West S. Longitudinal study of the microbiology of endemic trachoma. J Clin Microbiol. 1991 Aug;29(8):1593–1595. doi: 10.1128/jcm.29.8.1593-1595.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Taylor H. R., Young E., MacDonald A. B., Schachter J., Prendergast R. A. Oral immunization against chlamydial eye infection. Invest Ophthalmol Vis Sci. 1987 Feb;28(2):249–258. [PubMed] [Google Scholar]
  26. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Watkins N. G., Hadlow W. J., Moos A. B., Caldwell H. D. Ocular delayed hypersensitivity: a pathogenetic mechanism of chlamydial-conjunctivitis in guinea pigs. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7480–7484. doi: 10.1073/pnas.83.19.7480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Williams D. M., Byrne G. I., Grubbs B., Marshal T. J., Schachter J. Role in vivo for gamma interferon in control of pneumonia caused by Chlamydia trachomatis in mice. Infect Immun. 1988 Nov;56(11):3004–3006. doi: 10.1128/iai.56.11.3004-3006.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Williams D. M., Magee D. M., Bonewald L. F., Smith J. G., Bleicker C. A., Byrne G. I., Schachter J. A role in vivo for tumor necrosis factor alpha in host defense against Chlamydia trachomatis. Infect Immun. 1990 Jun;58(6):1572–1576. doi: 10.1128/iai.58.6.1572-1576.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zhong G. M., Peterson E. M., Czarniecki C. W., Schreiber R. D., de la Maza L. M. Role of endogenous gamma interferon in host defense against Chlamydia trachomatis infections. Infect Immun. 1989 Jan;57(1):152–157. doi: 10.1128/iai.57.1.152-157.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES