Polymerase chain reaction detection of Haemophilus ducreyi DNA.
OBJECTIVE
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.


CONCLUSIONS
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.
Roesel
D J
DJ
Center for AIDS Research, Stanford University, California 94305-5107, USA.
Gwanzura
L
L
Mason
P R
PR
Joffe
M
M
Katzenstein
D A
DA
D002602:Chancroid / Q000175:diagnosis; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005832:Genital Diseases, Male / Q000382:microbiology; D006191:Haemophilus ducreyi / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity; D014456:Ulcer / Q000382:microbiology
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S