Polymerase chain reaction detection of Haemophilus ducreyi DNA

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. (Sex Transm Inf 1998;74:63-65)

Treponema Infection Associated With Genital Ulceration in Wild Baboons

“…The PCRs targeting herpesvirus were positive in 23 of 26 tested individuals (Table 1): 19 clinically affected (17 of those PCR positive for T pallidum) and 7 clinically nonaffected (2 of those PCR positive for T pallidum). The PCRs revealed a variety of sequences, ranging from only 1 animal infected with 16S rRNA AAC GCG AAG AAC CTT ACC TG 56 GAC GGG CGG TGT GTA C Klebsiella granulomatis 8 phoE CTA TGA CAG CAA GGA TGG CGA/ 58 CAG ACC GAA GTC GAA CTG ATA CTG Haemophilus ducreyi 6,40,42 16S rRNA CAA GTC GAA CGG TAG CAC GAA G/ 58 TCA TCT CTG AGT TCT TCT ATG 16S rRNA (nested) 6 TTC TGT GAC TAA CGT CAA TCA ATT TTG/ 58 TCG GAT TAA AGG GTG GGA CCT T Pan-herpesvirus 13 dPol GCG GCC GAT GGT CGT GAC Treponema pallidum tpF1 (tp1038) 39 GAA AAA AAT ACC ACA GCA CCG C/ 57 CAA TGC CGT AGA TGT GCC AGT G tpF1 (tp1038) (nested) 39 CGT GCC ATT GCT GCT ATC T/ 56 TGC CGT AGA TGT GCC AGT G gpd (tp0257/glpQ) 7 AAG AAC TTT CCC TCC TCC GTG C/ 55 CGT TTG ATA CGC TTC AGC TCG/ gpd (tp0257/glpQ) (nested) 7 GTG GGT TGG AAC AGA CAA CC/ 55 CGT TTG CAC ATA CAC TAG ATC C deoD (tp0170/pfs) 10 GGT TAC CAG AAA GGG CGT ATT CC/ 55 CGA CCA TTA CAC GAC CAT C deoD (tp0170/pfs) (nested) 10 TAA TGG CGT CCC CTT TGT TA/ 55 GAA GCC ACT ACC GAT GTG C tprl 25 CGT CAC CCT CTC CTG GTA GT 60 ATC CCT CGC CTG TAA ACT GA tprl (nested) 25 GTG AGA GGA GGG GGA GTG A 55 CAC CAT TGG AAA GGA AGG AG polA 33 CGT CTG GTC GAT GTG CAA ATG AGT G/ 65 TGC ACA TGT ACA CTG AGT TGA CTC GG polA 31,32 AGG ATC CGG CAT ATG TCC AA/ 60 GTG AGC GTC TCA TCA TTC CAA A 6FAM-ATG CAC CAG CTT CGA-BBQ (probe) Human (housekeeping gene) c-myc 23 GCC AGA GGA GGA ACG AGC T/ 60 GGG CCT TTT CAT TGT TTT CCA 6FAM-TGC CCT GCG TGA CCA GAT CC-BBQ (probe) I, inosine. cercopithecine herpesvirus 16 (herpesvirus papio 2, case No.…”

Diagnosing Genital Ulcer Disease in a Clinic for Sexually Transmitted Diseases in Amsterdam, The Netherlands

“…The primers for H. ducreyi are located in the 16S rRNA gene and have, in part, been published (18,20). Outer primers, with a fragment length of 960 bp, were HD07 (5Ј CAA.GTC.GAA.CGG.TAG.CAC.GAA.G) and HD roe2 (5Ј TCA.TCT .CTG.AGT.TCT.TCT.ATG).…”

The use of molecular techniques for the diagnosis and epidemiologic study of sexually transmitted infections

“…Studies using panels of unrelated bacterial, fungal, and viral DNA showed that these assays seemed to specifically amplify or detect H. ducreyi-specific sequences. The performance of these assays has been determined by using men with a clinical diagnosis of chancroid [49,50,52], men with GUD [51], or consecutive patients (men alone or men and women) with GUD [43••,44-48,50]. The results of these studies are summarized in Table 3.…”