Skip to main content
NCBI home page
Epidemiology and Infection logoLink to Epidemiology and Infection
. 1993 Apr;110(2):289–295. doi: 10.1017/s0950268800068229

Non-radioactive ribotyping of Haemophilus ducreyi using a digoxigenin labelled cDNA probe.

T J Brown 1, C A Ison 1
PMCID: PMC2272269  PMID: 8386096

Abstract

Haemophilus ducreyi, the causal organism of chancroid, has increased in significance recently due to its association with HIV transmission. Most previous typing systems have exploited phenotypic characteristics. Detection of ribosomal RNA cistrons, ribotyping, was successfully developed to examine H. ducreyi, but required the use of 32P. We have used digoxigenin to define ribotypes from 30 strains of H. ducreyi from diverse geographical locations. This was achieved by agarose gel electrophoresis of restriction enzyme (RE) digested DNA extracts. These extracts were vacublotted onto nylon membrane and probed using digoxigenin labelled complementary DNA probe, prepared from Escherichia coli 16S and 23S ribosomal RNA. From 19 REs tested, Ava II, Hinc II, Bgl II and BstE II gave clear ribotypes. The ribotypes of BstE II and Bgl II used together gave the highest index of discrimination (D = 0.95), 16 types, and showed good reproducibility. This non-radioactive method demonstrates the three important features of a typing system; discrimination, typability and reproducibility.

Full text

PDF
289

Images in this article

293Fig. 1
on p.293

Selected References

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

  1. Albritton W. L. Biology of Haemophilus ducreyi. Microbiol Rev. 1989 Dec;53(4):377–389. doi: 10.1128/mr.53.4.377-389.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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 Nov;26(11):2465–2466. doi: 10.1128/jcm.26.11.2465-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Korting H. C., Abeck D., Johnson A. P., Ballard R. C., Taylor-Robinson D., Braun-Falco O. Lectin typing of Haemophilus ducreyi. Eur J Clin Microbiol Infect Dis. 1988 Oct;7(5):678–680. doi: 10.1007/BF01964253. [DOI] [PubMed] [Google Scholar]
  4. Morse S. A. Chancroid and Haemophilus ducreyi. Clin Microbiol Rev. 1989 Apr;2(2):137–157. doi: 10.1128/cmr.2.2.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Odumeru J. A., Ronald A. R., Albritton W. L. Characterization of cell proteins of Haemophilus ducreyi by polyacrylamide gel electrophoresis. J Infect Dis. 1983 Oct;148(4):710–714. doi: 10.1093/infdis/148.4.710. [DOI] [PubMed] [Google Scholar]
  6. Piot P., Laga M. Genital ulcers, other sexually transmitted diseases, and the sexual transmission of HIV. BMJ. 1989 Mar 11;298(6674):623–624. doi: 10.1136/bmj.298.6674.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Rossau R., Duhamel M., Jannes G., Decourt J. L., Van Heuverswyn H. The development of specific rRNA-derived oligonucleotide probes for Haemophilus ducreyi, the causative agent of chancroid. J Gen Microbiol. 1991 Feb;137(2):277–285. doi: 10.1099/00221287-137-2-277. [DOI] [PubMed] [Google Scholar]
  8. Sarafian S. K., Johnson S. R., Thomas M. L., Knapp J. S. Novel plasmid combinations in Haemophilus ducreyi isolates from Thailand. J Clin Microbiol. 1991 Oct;29(10):2333–2334. doi: 10.1128/jcm.29.10.2333-2334.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sarafian S. K., Woods T. C., Knapp J. S., Swaminathan B., Morse S. A. Molecular characterization of Haemophilus ducreyi by ribosomal DNA fingerprinting. J Clin Microbiol. 1991 Sep;29(9):1949–1954. doi: 10.1128/jcm.29.9.1949-1954.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Slootmans L., Vanden Berghe D. A., Piot P. Typing Haemophilus ducreyi by indirect immunofluorescence assay. Genitourin Med. 1985 Apr;61(2):123–126. doi: 10.1136/sti.61.2.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Stull T. L., LiPuma J. J., Edlind T. D. A broad-spectrum probe for molecular epidemiology of bacteria: ribosomal RNA. J Infect Dis. 1988 Feb;157(2):280–286. doi: 10.1093/infdis/157.2.280. [DOI] [PubMed] [Google Scholar]
  12. Van Dyck E., Piot P. Enzyme profile of Haemophilus ducreyi strains isolated on different continents. Eur J Clin Microbiol. 1987 Feb;6(1):40–43. doi: 10.1007/BF02097188. [DOI] [PubMed] [Google Scholar]

Articles from Epidemiology and Infection are provided here courtesy of Cambridge University Press

RESOURCES