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. 1989 Oct;27(10):2258–2262. doi: 10.1128/jcm.27.10.2258-2262.1989

Detection of leptospires in urine by polymerase chain reaction.

G J Van Eys 1, C Gravekamp 1, M J Gerritsen 1, W Quint 1, M T Cornelissen 1, J T Schegget 1, W J Terpstra 1
PMCID: PMC267006  PMID: 2584377

Abstract

Primers for polymerase chain reaction (PCR) were synthesized from clones derived from a Leptospira hardjo (type hardjobovis) library. One pair of synthetic oligonucleotide primers was selected for further analysis. Under experimental conditions an amplification was obtained with DNA of Leptospira interrogans of some serovars belonging to serogroup sejroe. However, very little or no amplification was observed with DNA from other serovars of this group. No amplification was observed with DNA from other serogroups, other bacteria, or eucaryotic organisms. Cattle urine, seeded with hardjobovis, was processed in several ways and subsequently subjected to PCR. Boiling of the samples or treatment with detergents appeared to be most effective. Urine samples containing fewer than 10 leptospires gave a positive result in the PCR assay. Twenty urine samples obtained from a slaughterhouse or farm cows were investigated using the PCR assay, culture isolation, dot and quick blot hybridization, and serological tests. This comparative study suggests that amplification by PCR may be a valuable method for the detection of leptospires in cattle urine.

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Selected References

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

  1. Demmler G. J., Buffone G. J., Schimbor C. M., May R. A. Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification. J Infect Dis. 1988 Dec;158(6):1177–1184. doi: 10.1093/infdis/158.6.1177. [DOI] [PubMed] [Google Scholar]
  2. Johnson R. C., Harris V. G. Differentiation of pathogenic and saprophytic letospires. I. Growth at low temperatures. J Bacteriol. 1967 Jul;94(1):27–31. doi: 10.1128/jb.94.1.27-31.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. LeFebvre R. B. DNA probe for detection of the Leptospira interrogans serovar hardjo genotype hardjo-bovis. J Clin Microbiol. 1987 Nov;25(11):2236–2238. doi: 10.1128/jcm.25.11.2236-2238.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Marshall R. B., Wilton B. E., Robinson A. J. Identification of Leptospira serovars by restriction-endonuclease analysis. J Med Microbiol. 1981 Feb;14(1):163–166. doi: 10.1099/00222615-14-1-163. [DOI] [PubMed] [Google Scholar]
  6. Ou C. Y., Kwok S., Mitchell S. W., Mack D. H., Sninsky J. J., Krebs J. W., Feorino P., Warfield D., Schochetman G. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science. 1988 Jan 15;239(4837):295–297. doi: 10.1126/science.3336784. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Shibata D. K., Arnheim N., Martin W. J. Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction. J Exp Med. 1988 Jan 1;167(1):225–230. doi: 10.1084/jem.167.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  12. Terpstra W. J., Schoone G. J., Ligthart G. S., ter Schegget J. Detection of Leptospira interrogans in clinical specimens by in situ hybridization using biotin-labelled DNA probes. J Gen Microbiol. 1987 Apr;133(4):911–914. doi: 10.1099/00221287-133-4-911. [DOI] [PubMed] [Google Scholar]
  13. Terpstra W. J., Schoone G. J., ter Schegget J. Detection of leptospiral DNA by nucleic acid hybridisation with 32P- and biotin-labelled probes. J Med Microbiol. 1986 Aug;22(1):23–28. doi: 10.1099/00222615-22-1-23. [DOI] [PubMed] [Google Scholar]
  14. Thiermann A. B., Handsaker A. L., Foley J. W., White F. H., Kingscote B. F. Reclassification of North American leptospiral isolates belonging to serogroups Mini and Sejroe by restriction endonuclease analysis. Am J Vet Res. 1986 Jan;47(1):61–66. [PubMed] [Google Scholar]
  15. Thiermann A. B., Handsaker A. L., Moseley S. L., Kingscote B. New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki. J Clin Microbiol. 1985 Apr;21(4):585–587. doi: 10.1128/jcm.21.4.585-587.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Van Eys G. J., Zaal J., Schoone G. J., Terpstra W. J. DNA hybridization with hardjobovis-specific recombinant probes as a method for type discrimination of Leptospira interrogans serovar hardjo. J Gen Microbiol. 1988 Mar;134(3):567–574. doi: 10.1099/00221287-134-3-567. [DOI] [PubMed] [Google Scholar]
  17. Wong C., Dowling C. E., Saiki R. K., Higuchi R. G., Erlich H. A., Kazazian H. H., Jr Characterization of beta-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA. 1987 Nov 26-Dec 2Nature. 330(6146):384–386. doi: 10.1038/330384a0. [DOI] [PubMed] [Google Scholar]
  18. Zuerner R. L., Bolin C. A. Repetitive sequence element cloned from Leptospira interrogans serovar hardjo type hardjo-bovis provides a sensitive diagnostic probe for bovine leptospirosis. J Clin Microbiol. 1988 Dec;26(12):2495–2500. doi: 10.1128/jcm.26.12.2495-2500.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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