Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1991 Aug;29(8):1711–1718. doi: 10.1128/jcm.29.8.1711-1718.1991

Use of polymerase chain reaction and rabbit infectivity testing to detect Treponema pallidum in amniotic fluid, fetal and neonatal sera, and cerebrospinal fluid.

E Grimprel 1, P J Sanchez 1, G D Wendel 1, J M Burstain 1, G H McCracken Jr 1, J D Radolf 1, M V Norgard 1
PMCID: PMC270188  PMID: 1761693

Abstract

The diagnosis of congenital syphilis continues to pose a difficult clinical challenge. Because the serodiagnosis of congenital syphilis has significant limitations, the direct detection of Treponema pallidum in suspect neonatal tissues or body fluids represents a desirable alternate diagnostic strategy. We developed and applied the polymerase chain reaction (PCR) for the detection of T. pallidum in clinical material relevant to the diagnosis of congenital syphilis but which typically contain factors inhibitory for the PCR. Four methods of specimen processing were examined to circumvent PCR inhibition; clinical materials included amniotic fluids, neonatal sera, and neonatal cerebrospinal fluids. The PCR was 100% specific for T. T. pallidum compared with the sensitive rabbit infectivity test (RIT) for all clinical materials tested. For amniotic fluids, the PCR was 100% sensitive when correlated with the RIT but had a lesser sensitivity when applied to sera or cerebrospinal fluids, which typically contain few treponemes. The combined sensitivity of the PCR for all clinical samples was 78%. Positive PCR results also were obtained among some clinical specimens for which RIT was not performed; these results correlated well with either stigmata or risk factors for congenital syphilis. The combined results suggest that the PCR can be a useful adjunct to the diagnosis and clinical management of congenital syphilis and that it will provide a valuable tool for investigations of the pathogenesis of the disorder.

Full text

PDF
1711

Images in this article

Selected References

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

  1. Ackerman B. D. Congenital syphilis: observations on laboratory diagnosis of intrauterine infection. J Pediatr. 1969 Mar;74(3):459–462. doi: 10.1016/s0022-3476(69)80204-0. [DOI] [PubMed] [Google Scholar]
  2. Alford C. A., Jr, Foft J. W., Blankenship W. J., Cassady G., Benton J. W., Jr Subclinical central nervous system disease of neonates: a prospective study of infants born with increased levels of IgM. J Pediatr. 1969 Dec;75(6):1167–1178. doi: 10.1016/s0022-3476(69)80372-0. [DOI] [PubMed] [Google Scholar]
  3. Alford C. A., Schaefer J., Blankenship W. J., Straumfjord J. V., Cassady G. A correlative immunologic, microbiologic and clinical approach to the diagnosis of acute and chronic infections in newborn infants. N Engl J Med. 1967 Aug 31;277(9):437–449. doi: 10.1056/NEJM196708312770901. [DOI] [PubMed] [Google Scholar]
  4. Bernet C., Garret M., de Barbeyrac B., Bebear C., Bonnet J. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction. J Clin Microbiol. 1989 Nov;27(11):2492–2496. doi: 10.1128/jcm.27.11.2492-2496.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brisson-Noël A., Gicquel B., Lecossier D., Lévy-Frébault V., Nassif X., Hance A. J. Rapid diagnosis of tuberculosis by amplification of mycobacterial DNA in clinical samples. Lancet. 1989 Nov 4;2(8671):1069–1071. doi: 10.1016/s0140-6736(89)91082-9. [DOI] [PubMed] [Google Scholar]
  6. Burg J. L., Grover C. M., Pouletty P., Boothroyd J. C. Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction. J Clin Microbiol. 1989 Aug;27(8):1787–1792. doi: 10.1128/jcm.27.8.1787-1792.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burstain J. M., Grimprel E., Lukehart S. A., Norgard M. V., Radolf J. D. Sensitive detection of Treponema pallidum by using the polymerase chain reaction. J Clin Microbiol. 1991 Jan;29(1):62–69. doi: 10.1128/jcm.29.1.62-69.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dobson S. R., Taber L. H., Baughn R. E. Recognition of Treponema pallidum antigens by IgM and IgG antibodies in congenitally infected newborns and their mothers. J Infect Dis. 1988 May;157(5):903–910. doi: 10.1093/infdis/157.5.903. [DOI] [PubMed] [Google Scholar]
  9. Dorfman D. H., Glaser J. H. Congenital syphilis presenting in infants after the newborn period. N Engl J Med. 1990 Nov 8;323(19):1299–1302. doi: 10.1056/NEJM199011083231902. [DOI] [PubMed] [Google Scholar]
  10. Fogel B. J., Tamer A. M., Fojaco R. M., Herrera J. A., Frank P. Immunologic response of the fetus in congenital syphilis. J Fla Med Assoc. 1969 Oct;56(10):777–779. [PubMed] [Google Scholar]
  11. Glover D. D., Winter C. A., Charles D., Larsen B. Diagnostic considerations in intra-amniotic syphilis. Sex Transm Dis. 1985 Jul-Sep;12(3):145–149. doi: 10.1097/00007435-198507000-00010. [DOI] [PubMed] [Google Scholar]
  12. Grover C. M., Thulliez P., Remington J. S., Boothroyd J. C. Rapid prenatal diagnosis of congenital Toxoplasma infection by using polymerase chain reaction and amniotic fluid. J Clin Microbiol. 1990 Oct;28(10):2297–2301. doi: 10.1128/jcm.28.10.2297-2301.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hance A. J., Grandchamp B., Lévy-Frébault V., Lecossier D., Rauzier J., Bocart D., Gicquel B. Detection and identification of mycobacteria by amplification of mycobacterial DNA. Mol Microbiol. 1989 Jul;3(7):843–849. doi: 10.1111/j.1365-2958.1989.tb00233.x. [DOI] [PubMed] [Google Scholar]
  14. Hardy D. A., Arvin A. M., Yasukawa L. L., Bronzan R. N., Lewinsohn D. M., Hensleigh P. A., Prober C. G. Use of polymerase chain reaction for successful identification of asymptomatic genital infection with herpes simplex virus in pregnant women at delivery. J Infect Dis. 1990 Nov;162(5):1031–1035. doi: 10.1093/infdis/162.5.1031. [DOI] [PubMed] [Google Scholar]
  15. Hay P. E., Clarke J. R., Strugnell R. A., Taylor-Robinson D., Goldmeier D. Use of the polymerase chain reaction to detect DNA sequences specific to pathogenic treponemes in cerebrospinal fluid. FEMS Microbiol Lett. 1990 Mar 15;56(3):233–238. doi: 10.1111/j.1574-6968.1990.tb13943.x. [DOI] [PubMed] [Google Scholar]
  16. Hsu P. L., Chamberlain N. R., Orth K., Moomaw C. R., Zhang L. Q., Slaughter C. A., Radolf J. D., Sell S., Norgard M. V. Sequence analysis of the 47-kilodalton major integral membrane immunogen of Treponema pallidum. Infect Immun. 1989 Jan;57(1):196–203. doi: 10.1128/iai.57.1.196-203.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaufman R. E., Jones O. G., Blount J. H., Wiesner P. J. Questionnaire survey of reported early congenital syphilis: problems in diagnosis, prevention, and treatment. Sex Transm Dis. 1977 Oct-Dec;4(4):135–139. doi: 10.1097/00007435-197710000-00003. [DOI] [PubMed] [Google Scholar]
  18. Kaufman R. E., Olansky D. C., Wiesner P. J. The FTA-ABS (IgM) test for neonatal congenital syphilis: A critical review. J Am Vener Dis Assoc. 1974 Dec;1(2):79–84. [PubMed] [Google Scholar]
  19. Khan W. N., Ali R. V., Werthmann M., Ross S. Immunoglobulin M determinations in neonates and infants as an adjunct to the diagnosis of infection. J Pediatr. 1969 Dec;75(6):1282–1286. doi: 10.1016/s0022-3476(69)80384-7. [DOI] [PubMed] [Google Scholar]
  20. Kogan S. C., Doherty M., Gitschier J. An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. N Engl J Med. 1987 Oct 15;317(16):985–990. doi: 10.1056/NEJM198710153171603. [DOI] [PubMed] [Google Scholar]
  21. Korones S. B., Roane J. A., Gilkeson M. R., Lafferty W., Sever J. L. Neonatal IgM response to acute infection. J Pediatr. 1969 Dec;75(6):1261–1270. doi: 10.1016/s0022-3476(69)80382-3. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lewis L. L., Taber L. H., Baughn R. E. Evaluation of immunoglobulin M western blot analysis in the diagnosis of congenital syphilis. J Clin Microbiol. 1990 Feb;28(2):296–302. doi: 10.1128/jcm.28.2.296-302.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lukehart S. A., Hook E. W., 3rd, Baker-Zander S. A., Collier A. C., Critchlow C. W., Handsfield H. H. Invasion of the central nervous system by Treponema pallidum: implications for diagnosis and treatment. Ann Intern Med. 1988 Dec 1;109(11):855–862. doi: 10.7326/0003-4819-109-11-855. [DOI] [PubMed] [Google Scholar]
  25. Malloy D. C., Nauman R. K., Paxton H. Detection of Borrelia burgdorferi using the polymerase chain reaction. J Clin Microbiol. 1990 Jun;28(6):1089–1093. doi: 10.1128/jcm.28.6.1089-1093.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mascola L., Pelosi R., Blount J. H., Alexander C. E., Cates W., Jr Congenital syphilis revisited. Am J Dis Child. 1985 Jun;139(6):575–580. doi: 10.1001/archpedi.1985.02140080045030. [DOI] [PubMed] [Google Scholar]
  27. McCracken G. H., Jr, Hardy J. B., Chen T. C., Hoffman L. S., Gilkeson M. R., Sever J. L. Serum immunoglobulin levels in newborn infants. II. Survey of cord and follow-up sera from 123 infants with congenital rubella. J Pediatr. 1969 Mar;74(3):383–392. doi: 10.1016/s0022-3476(69)80195-2. [DOI] [PubMed] [Google Scholar]
  28. Miller M. J., Sunshine P. J., Remington J. S. Quantitation of cord serum IgM and IgA as a screening procedure to detect congenital infection: results in 5,006 infants. J Pediatr. 1969 Dec;75(6):1287–1291. doi: 10.1016/s0022-3476(69)80385-9. [DOI] [PubMed] [Google Scholar]
  29. Moser D. R., Kirchhoff L. V., Donelson J. E. Detection of Trypanosoma cruzi by DNA amplification using the polymerase chain reaction. J Clin Microbiol. 1989 Jul;27(7):1477–1482. doi: 10.1128/jcm.27.7.1477-1482.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Srinivasan G., Ramamurthy R. S., Bharathi A., Voora S., Pildes R. S. Congenital syphilis: a diagnostic and therapeutic dilemma. Pediatr Infect Dis. 1983 Nov-Dec;2(6):436–441. [PubMed] [Google Scholar]
  31. Sánchez P. J., McCracken G. H., Jr, Wendel G. D., Olsen K., Threlkeld N., Norgard M. V. Molecular analysis of the fetal IgM response to Treponema pallidum antigens: implications for improved serodiagnosis of congenital syphilis. J Infect Dis. 1989 Mar;159(3):508–517. doi: 10.1093/infdis/159.3.508. [DOI] [PubMed] [Google Scholar]
  32. Turner T. B., Hardy P. H., Newman B. Infectivity tests in syphilis. Br J Vener Dis. 1969 Sep;45(3):183–195. doi: 10.1136/sti.45.3.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wendel G. D., Maberry M. C., Christmas J. T., Goldberg M. S., Norgard M. V. Examination of amniotic fluid in diagnosing congenital syphilis with fetal death. Obstet Gynecol. 1989 Dec;74(6):967–970. [PubMed] [Google Scholar]

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

RESOURCES