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Sex Transm Infect doi:10.1136/sti.2008.034207

Evaluation of multiplex real-time PCR for detection of H. ducreyi, T. pallidum, HSV-1, and HSV-2 in the diagnosis of genital ulcer disease in Rakai District, Uganda

  1. Tara R Suntoke (tsuntoke{at}yahoo.com)
  1. NIAID/NIH, United States
    1. Andrew Hardick
    1. Johns Hopkins Medical Institutions, United States
      1. Aaron A. R. Tobian
      1. Johns Hopkins Medical Institutions, United States
        1. Bryan Mposa
        1. Rakai Health Sciences Program, Uganda
          1. Oliver Laeyendecker
          1. NIAID/NIH and Johns Hopkins Medical Institutions, United States
            1. David Serwadda
            1. Makerere University, Uganda
              1. Pius Opendi
              1. Rakai Health Sciences Program, Uganda
                1. Charlotte A Gaydos
                1. Johns Hopkins Medical Institutions, United States
                  1. Ronald H Gray
                  1. Johns Hopkins School of Public Health, United States
                    1. Maria J Wawer
                    1. Johns Hopkins School of Public Health, United States
                      1. Thomas C Quinn (tquinn{at}jhmi.edu)
                      1. NIAID/NIH and Johns Hopkins Medical Institutions, United States
                        1. Steven J Reynolds
                        1. NIAID/NIH and Johns Hopkins Medical Institutions, United States
                          • Published Online First 9 December 2008

                          Abstract

                          Objective: To develop a real-time PCR assay that reliably and accurately detects the predominant sexually transmitted aetiologic 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 utility 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/T. pallidum and HSV-1/HSV-2 in ulcer swabs from 100 persons with symptomatic genital ulcers in rural Rakai Uganda. Results were compared with syphilis, HSV-1 and HSV-2 serology.

                          Results: Of 100 GUD samples analyzed from 43 HIV-positive and 57 HIV-negative individuals, 71% were positive for one or more 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 US, 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.