1887

Journal of Medical Microbiology logo

Volume 46, Issue 2

Differentiation of species and serovars by PCR-restriction endonuclease analysis, arbitrarily primed PCR and low-stringency PCR Free

Abstract

Reference strains from 30 serovars representing seven species of and 48 recent isolates from human patients, dogs and rats, were characterised by polymerase chain reaction-restriction endonuclease analysis (PCR-REA), arbitrarily primed PCR (AP-PCR) and low stringency PCR (LS-PCR). PCR-REA analysis yielded seven groups among 29 serovars of pathogenic ; the non-pathogenic serovar was not amplified with the primer pairs studied. AP-PCR and LS-PCR fingerprinting resulted in 25 and 21 distinct profiles, respectively, among the 30 reference strains. The results of the three PCR-based techniques were highly concordant and were in general agreement with those from previous DNA studies, confirming the high level of polymorphism among species and serovars, and supported the concept of the serovar as the basic taxonomic unit of leptospiral classification. Results of the PCR-based typing methods for 11 randomised leptospiral strains, 36 clinical isolates from human patients and dogs and 12 survey isolates from trapped rats agreed with those from serological identification. With one exception, isolates of the same serovar gave identical profiles irrespective of the source. AP-PCR and LS-PCR are simple to perform and interpret, and appear to be useful for characterising isolates of spp. for diagnostic and epidemiological purposes.

  • Received:
  • Published Online:
© 1997 The Pathological Society of Great Britain and Ireland
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-46-2-173
1997-02-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jmm/46/2/medmicro-46-2-173.html?itemId=/content/journal/jmm/10.1099/00222615-46-2-173&mimeType=html&fmt=ahah

References

  1. Kmety E., Dikken H. Revised list of Leptospira serovars accepted by the sub-committee on the taxonomy of Leptospira at the Manchester meeting. 6/7 September 1986 Groningen: University Press;1988
  2. Kmety E., Dikken H. Classification of the species Leptospira interrogans and history of its serovars. Groningen: University Press; 19931–8 89–96
    [Google Scholar]
  3. Ramadass P., Jarvis B. D. W., Corner R. J., Penny D., Marshall R. B. Genetic characterization of pathogenic Leptospira species by DNA hybridization. Int J Syst Bacteriol 1992; 42:215–219
     [Google Scholar]
  4. Yasuda P. H., Stiegerwalt A. G., Sulzer K. R., Kaufmann A. F., Rogers F., Brenner D. J. Deoxyribonucleic acid relatedness between serogroups and serovars in the family Leptospiraceae with proposals for seven new Leptospira species. Int J Syst Bacteriol 1987; 37:407–415
     [Google Scholar]
  5. Faine S. (ed) Guidelines for the control of leptospirosis. WHO Offset Publication No. 67 Geneva: World Health Organization; 1982; 2043
     [Google Scholar]
  6. Michna S. W. Leptospirosis. Vet Rec 1970; 86:484–496
     [Google Scholar]
  7. Turner L. H. Leptospirosis II. Serology. Trans R Soc Trop Med Hyg 1968; 62:880–899
     [Google Scholar]
  8. Everard J. D., Everard C. O. R. Leptospirosis in the Caribbean. Rev Med Microbiol 1993; 4:114–122
     [Google Scholar]
  9. Everard C. O. R., Edwards C. N., Everard J. D., Carrington D. G. A twelve-year study of leptospirosis on Barbados. Eur J Epidemiol 1995; 11:311–320
     [Google Scholar]
  10. Marshall R. B., Wilton B. E., Robinson A. J. Identification of Leptospira serovars by restriction-endonuclease analysis. J Med Microbiol 1981; 14:163–166
     [Google Scholar]
  11. LeFebvre R. B. DNA probe for detection of the Leptospira interrogans serovar hardjo genotype hardjo-bovis. J Clin Microbiol 1987; 25:2236–2238
     [Google Scholar]
  12. van Eys G. J. J. M., 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; 134:567–574
     [Google Scholar]
  13. Perolat P., Grimont F., Regnault B. rRNA gene restriction patterns of Leptospira: a molecular typing system. Res Microbiol 1990; 141:159–171
     [Google Scholar]
  14. Zuerner R. L., Bolin C. A. Nucleic acid probe characterizes Leptospira interrogans serovars by restriction fragment length polymorphisms. Vet Microbiol 1990; 24:355–366
     [Google Scholar]
  15. Herrmann J. L., Bellenger E., Perolat P., Baranton G., Saint Girons I. Pulsed-field gel electrophoresis of Not I digests of leptospiral DNA: a new rapid method of serovar identification. J Clin Microbiol 1992; 30:1696–1702
     [Google Scholar]
  16. Hookey J. V. Characterization of Leptospiraceae by 16S DNA restriction fragment length polymorphisms. J Gen Microbiol 1993; 139:1681–1689
     [Google Scholar]
  17. Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 1990; 18:7213–7218
     [Google Scholar]
  18. Williams J. G. K., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 1990; 18:6531–6535
     [Google Scholar]
  19. Elaichouni A., Verschraegen G., Claeys G., Devleeschouwer M., Godard C., Vaneechoutte M. Pseudomonas aeruginosa serotype O12 outbreak studied by arbitrary primer PCR. J Clin Microbiol 1994; 32:666–671
     [Google Scholar]
  20. Corney B. G., Colley J., Djordjevic S. P., Whittington R., Graham G. C. Rapid identification of some Leptospira isolates from cattle by random amplified polymorphic DNA fingerprinting. J Clin Microbiol 1993; 31:2927–2932
     [Google Scholar]
  21. Levett P. N., Bennett P., O’Donaghue K., Bowker K., Reeves D., MacGowan A. Relapsed infection due to Listeria monocytogenes confirmed by random amplified polymorphic DNA (RAPD) analysis. J Infect 1993; 27:205–207
     [Google Scholar]
  22. Welsh J., Pretzman C., Postic D., Saint Girons I., Baranton G., McClelland M. Genomic fingerprinting by arbitrarily primed polymerase chain reaction resolves Borrelia burgdorferi into three distinct phyletic groups. Int J Syst Bacteriol 1992; 42:370–377
     [Google Scholar]
  23. Gravekamp C., van de Kemp H., Franzen M. Detection of seven species of pathogenic leptospires by PCR using two sets of primers. J Gen Microbiol 1993; 139:1691–1700
     [Google Scholar]
  24. Ellinghausen H. C., McCullough W. G. Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and a medium of bovine albumin and polysorbate 80. Am J Vet Res 1965; 26:45–51
     [Google Scholar]
  25. Johnson R. C., Harris V. G. Differentiation of pathogenic and saprophytic leptospires. 1. Growth at low temperatures. J Bacteriol 1967; 94:27–31
     [Google Scholar]
  26. Brown P. D., Gravekamp C., Carrington D. G. Evaluation of the polymerase chain reaction for early diagnosis of leptospirosis. J Med Microbiol 1995; 43:110–114
     [Google Scholar]
  27. Hunter P. R., Gaston M. A. Numerical index of the discriminatory ability of typing systems: an application of Simpson’s index of diversity. J Clin Microbiol 1988; 26:2465–2466
     [Google Scholar]
  28. Savio M. L., Rossi C., Fusi P., Tagliablue S., Pacciarini M. L. Detection and identification of Leptospira interrogans serovars by PCR coupled with restriction endonuclease analysis of amplified DNA. J Clin Microbiol 1994; 32:935–941
     [Google Scholar]
  29. Damas de Caballero O. L. S., Dias Neto E., Koury M. C., Romanha A. J., Simpson A. J. G. Low-stringency PCR with diagnostically useful primers for identification of Leptospira serovars. J Clin Microbiol 1994; 32:1369–1372
     [Google Scholar]
  30. Herrmann J. L. Genomic techniques for identification of Leptospira strains. Pathol Biol 1993; 41:943–950
     [Google Scholar]
  31. Tamai T., Sada E., Kobayashi Y. Restriction endonuclease DNA analysis of Leptospira interrogans serovars Icterohaemorrhagiae and Copenhageni . Microbiol Immunol 1988; 32:887–894
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-46-2-173
Loading
Differentiation of Leptospira species and serovars by PCR-restriction endonuclease analysis, arbitrarily primed PCR and low-stringency PCR
J. Med. Microbiol. 46, 173 (1997); https://doi.org/10.1099/00222615-46-2-173
/content/journal/jmm/10.1099/00222615-46-2-173
/content/journal/jmm/10.1099/00222615-46-2-173
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error