Hostname: page-component-586b7cd67f-t8hqh Total loading time: 0 Render date: 2024-11-30T21:02:01.507Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  19 October 2009

T. J. Brown  and
C. A. Ison
T. J. Brown
Affiliation:
Department of Medical Microbiology, St Mary's Hospital Medical School, Paddington, London W2 1PG
C. A. Ison
Affiliation:
Department of Medical Microbiology, St Mary's Hospital Medical School, Paddington, London W2 1PG
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

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 phenotypie 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 nonradioactive method demonstrates the three important features of a typing system: discrimination, typability and reproducibility.

Type
Research Article
Information
Epidemiology & Infection , Volume 110 , Issue 2 , April 1993 , pp. 289 - 295
Copyright
Copyright © Cambridge University Press 1993

References

REFERENCES

1.Albritton, WL. Biology of Haemophilus ducreyi. Microbiol Rev 1989; 53: 377–89.CrossRefGoogle ScholarPubMed
2.Morse, SA. Chancroid and Haemophilus ducreyi. Clin Microbiol Rev 1989; 2: 137–57.CrossRefGoogle ScholarPubMed
3.Piot, P, Laga, M. Genital ulcers, other sexually transmitted diseases, and the sexual transmission of HIV. Brit Med J 1989; 298: 623–4.CrossRefGoogle ScholarPubMed
4.Odumeru, JA, Ronald, AR, Albritton, WL. Characterization of cell proteins of Haemophilus ducreyi by polyacrylamide gel electrophoresis. J Infect Dis 1983; 148: 710–4.CrossRefGoogle ScholarPubMed
5.Van Dyke, E, Piot, P. Enzyme profile of Ilaemophilus ducreyi strains isolated on different continents. Eur J Clin Microbiol 1987; 6: 40–3.CrossRefGoogle Scholar
6.Korting, HC, Abeck, D, Johnson, AP, Ballard, RC, Taylor-Robinson, D. Lectin typing of Haemophilus ducreyi. Eur J Clin Microbiol Infect Dis 1988; 7: 678–80.CrossRefGoogle ScholarPubMed
7.Slootmans, L, Vanden Berghe, DA, Piot, P. Typing Haemophilus ducreyi by indirect immunofluorescence assay. Genitourin Med 1985; 61: 123–6.Google ScholarPubMed
8.Sarafian, SK, Johnson, SR, Thomas, ML, Knapp, JS. Novel plasmid combinations in Haemophilus ducreyi isolates from Thailand. J Clin Microbiol 1991; 29: 2333–4.CrossRefGoogle ScholarPubMed
9.Stull, TL, LiPuma, JJ, Edlind, TD. A broad-spectrum probe for molecular epidemiology of bacteria: Ribosoinal RNA. J Infect Dis 1988; 157: 280–6.CrossRefGoogle ScholarPubMed
10.Sarafian, SK, Woods, TC, Knapp, JS, Swaminatban, B, Morse, SA. Molecular characterization of Haemophilus ducreyi by ribosoinal DNA fingerprinting. J Clin Mierobiol 1991: 29: 1949–54.CrossRefGoogle ScholarPubMed
11.Pitcher, DG, Owen, RJ, Dyal, P, Beck, A. Synthesis of a biotinylated DNA probe to detect ribosomal RNA cistrons in Providencia stuarlii. FEMS Mierobiol Lett 1987: 48: 283–7.CrossRefGoogle Scholar
12.Krieg, RN, ed. Berg's manual of systematic bacteriology, vol 1. Baltimore: Williams and Wilkins, 1984: 558–6.Google Scholar
13.Sambrook, J, Fritsch, EF, Maniatis, T. Molecular cloning, a laboratory manual. 2nd edCold Spring Harbor: Cold Spring Harbor Laboratory Press 1989: E3–17.Google Scholar
14.Hunter, PR, Gaston, MA. Numerical index of the discriminatory ability of typing systems: an application of Simpson's Index of Diversity. J Clin Mierobiol 1988: 26: 2465–6.Google ScholarPubMed
15.Rossau, R, Duhamel, M, Jannes, G, Decourt, JL, Van Heuverswyn, H. The development of specific rRNA-derived oligonucleotide probes for Haemophilus ducreyi. the causative agent of chancroid. J Gen Mierobiol 1991; 137: 277–85.Google ScholarPubMed