This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Liu, H.
Right arrow Articles by Steiner, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Liu, H.
Right arrow Articles by Steiner, B.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 2001, p. 1941-1946, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1941-1946.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene

Hsi Liu,* Berta Rodes, C.-Y. Chen, and Bret Steiner

Centers for Disease Control and Prevention, Atlanta, Georgia 30333

Received 14 December 2000/Returned for modification 18 January 2001/Accepted 21 February 2001

A sensitive and specific PCR method to detect Treponema pallidum in clinical specimens was developed. PCR primers were designed based on two unique features of the DNA polymerase I gene (polA). The first distinctive characteristic is that the region codes for a high cysteine content and has low homology with similar regions of DNA polymerase I gene from known microorganisms. The second unique feature is the presence of four insertions in the gene. PCR tests using primers designed on the basis these regions reacted with various pathogenic T. pallidum subspecies but did not react with nonpathogenic treponemal species or other spirochetes. An additional 59 species of bacteria and viruses, including those that cause genital ulcers, tested negative. This PCR method is extremely robust and sensitive. The detection limit is about 10 to 25 organisms when analyzed on gel. However, the analytic sensitivity can be increased by at least 1 log, to a detection limit of a single organism, when the ABI 310 Prism Genetic Analyzer is used to detect fluorescence-labeled amplicons. We further used this test in a clinical setting and compared the results with results from a previously reported multiplex-PCR test (for T. pallidum, Haemophilus ducreyi, and herpes simplex virus). We tested 112 genital ulcer specimens by the polA PCR, obtaining a sensitivity of 95.8% and a specificity of 95.7%. These results suggest that the polA PCR is applicable as a routine clinical diagnostic test for syphilis.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Rd., Mail Stop D13, Atlanta, GA 30333. Phone: (404) 639-3348. Fax: (404) 639-3976. E-mail: hcl6{at}cdc.gov.

Journal of Clinical Microbiology, May 2001, p. 1941-1946, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1941-1946.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Martin, I. E., Tsang, R. S. W., Sutherland, K., Tilley, P., Read, R., Anderson, B., Roy, C., Singh, A. E. (2009). Molecular Characterization of Syphilis in Patients in Canada: Azithromycin Resistance and Detection of Treponema pallidum DNA in Whole-Blood Samples versus Ulcerative Swabs. J. Clin. Microbiol. 47: 1668-1673 [Abstract] [Full Text]  
  • Matejkova, P., Flasarova, M., Zakoucka, H., Borek, M., Kremenova, S., Arenberger, P., Woznicova, V., Weinstock, G. M., Smajs, D. (2009). Macrolide treatment failure in a case of secondary syphilis: a novel A2059G mutation in the 23S rRNA gene of Treponema pallidum subsp. pallidum. J Med Microbiol 58: 832-836 [Abstract] [Full Text]  
  • Florindo, C., Reigado, V., Gomes, J. P., Azevedo, J., Santo, I., Borrego, M. J. (2008). Molecular Typing of Treponema pallidum Clinical Strains from Lisbon, Portugal. J. Clin. Microbiol. 46: 3802-3803 [Abstract] [Full Text]  
  • Behrhof, W, Springer, E, Brauninger, W, Kirkpatrick, C J, Weber, A (2008). PCR testing for Treponema pallidum in paraffin-embedded skin biopsy specimens: test design and impact on the diagnosis of syphilis. J. Clin. Pathol. 61: 390-395 [Abstract] [Full Text]  
  • Chakraborty, R., Luck, S. (2008). Syphilis is on the increase: the implications for child health. Arch. Dis. Child. 93: 105-109 [Full Text]  
  • Pandori, M. W., Gordones, C., Castro, L., Engelman, J., Siedner, M., Lukehart, S., Klausner, J. (2007). Detection of Azithromycin Resistance in Treponema pallidum by Real-Time PCR. Antimicrob. Agents Chemother. 51: 3425-3430 [Abstract] [Full Text]  
  • Zetola, N. M., Engelman, J., Jensen, T. P., Klausner, J. D. (2007). Syphilis in the United States: An Update for Clinicians With an Emphasis on HIV Coinfection. Mayo Clin Proc. 82: 1091-1102 [Abstract] [Full Text]  
  • Muller, M, Ewert, I, Hansmann, F, Tiemann, C, Hagedorn, H J, Solbach, W, Roider, J, Nolle, B, Laqua, H, Hoerauf, H (2007). Detection of Treponema pallidum in the vitreous by PCR. Br. J. Ophthalmol. 91: 592-595 [Abstract] [Full Text]  
  • Leslie, D. E., Azzato, F., Karapanagiotidis, T., Leydon, J., Fyfe, J. (2007). Development of a Real-Time PCR Assay To Detect Treponema pallidum in Clinical Specimens and Assessment of the Assay's Performance by Comparison with Serological Testing. J. Clin. Microbiol. 45: 93-96 [Abstract] [Full Text]  
  • Chen, C.-Y., Chi, K.-H., George, R. W., Cox, D. L., Srivastava, A., Rui Silva, M., Carneiro, F., Lauwers, G. Y., Ballard, R. C. (2006). Diagnosis of gastric syphilis by direct immunofluorescence staining and real-time PCR testing.. J. Clin. Microbiol. 44: 3452-3456 [Abstract] [Full Text]  
  • Morshed, M. G., Jones, H. D. (2006). Treponema pallidum macrolide resistance in BC. CMAJ 174: 349-349 [Full Text]  
  • Pope, V., Fox, K., Liu, H., Marfin, A. A., Leone, P., Sena, A. C., Chapin, J., Fears, M. B., Markowitz, L. (2005). Molecular Subtyping of Treponema pallidum from North and South Carolina. J. Clin. Microbiol. 43: 3743-3746 [Abstract] [Full Text]  
  • van Assen, S., Bakker, S. J. L. (2005). Did syphilis truly strike the kidneys this time?. Nephrol Dial Transplant 20: 1029-1031 [Full Text]  
  • Palmer, H M, Higgins, S P, Herring, A J, Kingston, M A (2003). Use of PCR in the diagnosis of early syphilis in the United Kingdom. Sex. Transm. Infect. 79: 479-483 [Abstract] [Full Text]  
  • Pillay, A., Liu, H., Ebrahim, S., Chen, C. Y., Lai, W., Fehler, G., Ballard, R. C., Steiner, B., Sturm, A. W., Morse, S. A. (2002). Molecular Typing of Treponema pallidum in South Africa: Cross-Sectional Studies. J. Clin. Microbiol. 40: 256-258 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»