National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States; Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Institute of Public Health, Makerere University, Kampala, Uganda; Rakai Health Sciences Program, Rakai, Uganda; Department of Population and Family Health Sciences, Bloomberg School of Public Health, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Division of Infectious Diseases, Johns Hopkins Medical Institutions, 855 N. Wolfe Street, Baltimore, MD 21205, United States
Suntoke, T.R., National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States; Hardick, A., Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Tobian, A.A.R., Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Mpoza, B., Rakai Health Sciences Program, Rakai, Uganda; Laeyendecker, O., National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Serwadda, D., Institute of Public Health, Makerere University, Kampala, Uganda; Opendi, P., Rakai Health Sciences Program, Rakai, Uganda; Gaydos, C.A., Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Gray, R.H., Department of Population and Family Health Sciences, Bloomberg School of Public Health, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Wawer, M.J., Department of Population and Family Health Sciences, Bloomberg School of Public Health, Johns Hopkins Medical Institutions, Baltimore, MD, United States; Quinn, T.C., National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States, Division of Infectious Diseases, Johns Hopkins Medical Institutions, 855 N. Wolfe Street, Baltimore, MD 21205, United States; Reynolds, S.J., National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, Division of Infectious Diseases, Johns Hopkins Medical Institutions, Baltimore, MD, United States
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-7). 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 (κ = 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.