Detection and discrimination of herpes simplex viruses, Haemophilus ducreyi, Treponema pallidum, and Calymmatobacterium (Klebsiella) granulomatis from genital ulcers.
BACKGROUND
Genital ulcer disease (GUD) is commonly caused by pathogens for which suitable therapies exist, but clinical and laboratory diagnoses may be problematic. This collaborative project was undertaken to address the need for a rapid, economical, and sensitive approach to the detection and diagnosis of GUD using noninvasive techniques to sample genital ulcers.


METHODS
The genital ulcer disease multiplex polymerase chain reaction (GUMP) was developed as an inhouse nucleic acid amplification technique targeting serious causes of GUD, namely, herpes simplex viruses (HSVs), H. ducreyi, Treponema pallidum, and Klebsiella species. In addition, the GUMP assay included an endogenous internal control. Amplification products from GUMP were detected by enzyme linked amplicon hybridization assay (ELAHA).


RESULTS
GUMP-ELAHA was sensitive and specific in detecting a target microbe in 34.3% of specimens, including 1 detection of HSV-1, three detections of HSV-2, and 18 detections of T. pallidum. No H. ducreyi has been detected in Australia since 1998, and none was detected here. No Calymmatobacterium (Klebsiella) granulomatis was detected in the study, but there were 3 detections during ongoing diagnostic use of GUMP-ELAHA in 2004 and 2005. The presence of C. granulomatis was confirmed by restriction enzyme digestion and nucleotide sequencing of the 16S rRNA gene for phylogenetic analysis.


CONCLUSIONS
GUMP-ELAHA permitted comprehensive detection of common and rare causes of GUD and incorporated noninvasive sampling techniques. Data obtained by using GUMP-ELAHA will aid specific treatment of GUD and better define the prevalence of each microbe among at-risk populations with a view to the eradication of chancroid and donovanosis in Australia.
Mackay
Ian M
IM
Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Queensland, Australia. ian.mackay@uq.edu.au
Harnett
Gerry
G
Jeoffreys
Neisha
N
Bastian
Ivan
I
Sriprakash
Kadaba S
KS
Siebert
David
D
Sloots
Theo P
TP
D002157:Calymmatobacterium / Q000302:isolation & purification*; D002602:Chancroid / Q000175:diagnosis*; D017931:DNA Primers; D003937:Diagnosis, Differential; D004756:Enterobacteriaceae Infections / Q000175:diagnosis*; D005260:Female; D005831:Genital Diseases, Female / Q000175:diagnosis / Q000382:microbiology / Q000821:virology; D005832:Genital Diseases, Male / Q000175:diagnosis / Q000382:microbiology / Q000821:virology; D006099:Granuloma / Q000175:diagnosis; D006191:Haemophilus ducreyi / Q000302:isolation & purification*; D006561:Herpes Simplex / Q000175:diagnosis*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D012749:Sexually Transmitted Diseases / Q000175:diagnosis*; D018139:Simplexvirus / Q000302:isolation & purification*; D013587:Syphilis / Q000175:diagnosis; D014210:Treponema pallidum / Q000302:isolation & purification*; D014456:Ulcer / Q000175:diagnosis
D017931:DNA Primers