Evaluation of multiplex real-time PCR for detection of Haemophilus ducreyi, Treponema pallidum, herpes simplex virus type 1 and 2 in the diagnosis of genital ulcer disease in the Rakai District, Uganda.
OBJECTIVE
To develop a real-time PCR assay that reliably and accurately detects the predominant sexually transmitted aetiological agents of genital ulcer disease (GUD) (Haemophilus ducreyi, Treponema pallidum and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2)) and to assess the use of real-time PCR diagnostic testing in a rural African field site.


METHODS
Two multiplex real-time PCR reactions were used to detect H ducreyi/and HSV-1/HSV-2 in ulcer swabs from 100 people with symptomatic genital ulcers in rural Rakai, Uganda. Results were compared with syphilis, HSV-1 and HSV-2 serology.


RESULTS
Of 100 GUD samples analysed from 43 HIV positive and 57 HIV negative individuals, 71% were positive for one or more sexually transmitted infection (STI) pathogens by real-time PCR (61% for HSV-2, 5% for T pallidum, 3% for HSV-1, 1% for H ducreyi and 1% for dual H ducreyi/HSV-2). The frequency of HSV in genital ulcers was 56% (32/57) in HIV negative individuals and 77% (33/43) in HIV positive individuals (p = 0.037). Assay reproducibility was evaluated by repeat PCR testing in the USA with 96% agreement (kappa = 0.85).


CONCLUSIONS
STI pathogens were detected in the majority of GUD swab samples from symptomatic patients in Rakai, Uganda, by real-time PCR. HSV-2 was the predominant cause of genital ulcers. Real-time PCR technology can provide sensitive, rapid and reproducible evaluation of GUD aetiology in a resource-limited setting.
Suntoke
T R
TR
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Hardick
A
A
Tobian
A A R
AA
Mpoza
B
B
Laeyendecker
O
O
Serwadda
D
D
Opendi
P
P
Gaydos
C A
CA
Gray
R H
RH
Wawer
M J
MJ
Quinn
T C
TC
Reynolds
S J
SJ
D000328:Adult; D015331:Cohort Studies; D005260:Female; D015658:HIV Infections / Q000150:complications; D006191:Haemophilus ducreyi / Q000302:isolation & purification*; D018259:Herpesvirus 1, Human / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012423:Rural Health; D012749:Sexually Transmitted Diseases / Q000175:diagnosis / Q000382:microbiology* / Q000821:virology; D014210:Treponema pallidum / Q000302:isolation & purification*; D014454:Uganda; D014456:Ulcer / Q000175:diagnosis / Q000382:microbiology* / Q000821:virology; D055815:Young Adult