Polymerase chain reaction detection of Haemophilus ducreyi DNA

Sex Transm Infect. 1998 Feb;74(1):63-5. doi: 10.1136/sti.74.1.63.

Abstract

Objectives: To develop a polymerase chain reaction (PCR) method to detect Haemophilus ducreyi DNA in cultured isolates and clinical material.

Methods: Primers specific to the H ducreyi 16s rRNA gene were synthesised. PCR conditions were optimised and products were verified by restriction endonuclease digestion and agarose gel electrophoresis.

Results: The method was able to detect all 28 H ducreyi strains tested; specificity was demonstrated using lysates of 12 related organisms. Applied to clinical samples from genital ulcer swabs obtained in Harare, Zimbabwe, H ducreyi DNA was detected in repeated assays in 35 clinical samples.

Conclusion: PCR amplification using primers from the 16s rRNA gene may be a useful alternative to culture for the detection of H ducreyi and the diagnosis of chancroid.

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chancroid / diagnosis
  • DNA Primers
  • DNA, Bacterial / isolation & purification*
  • Electrophoresis, Agar Gel
  • Genital Diseases, Male / microbiology
  • Haemophilus ducreyi / genetics
  • Haemophilus ducreyi / isolation & purification*
  • Humans
  • Male
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 16S / analysis
  • Sensitivity and Specificity
  • Ulcer / microbiology

Substances

  • DNA Primers
  • DNA, Bacterial
  • RNA, Ribosomal, 16S