title: 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.
abstract: 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.
authors.affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: T R
authors.initials: TR
authors.lastname: Suntoke
authors.forename: A
authors.initials: A
authors.lastname: Hardick
authors.forename: A A R
authors.initials: AA
authors.lastname: Tobian
authors.forename: B
authors.initials: B
authors.lastname: Mpoza
authors.forename: O
authors.initials: O
authors.lastname: Laeyendecker
authors.forename: D
authors.initials: D
authors.lastname: Serwadda
authors.forename: P
authors.initials: P
authors.lastname: Opendi
authors.forename: C A
authors.initials: CA
authors.lastname: Gaydos
authors.forename: R H
authors.initials: RH
authors.lastname: Gray
authors.forename: M J
authors.initials: MJ
authors.lastname: Wawer
authors.forename: T C
authors.initials: TC
authors.lastname: Quinn
authors.forename: S J
authors.initials: SJ
authors.lastname: Reynolds
mesh_terms: 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