SIMULTANEOUS DETECTION OF 3 COMMON SEXUALLY-TRANSMITTED AGENTS BY POLYMERASE CHAIN-REACTION

OBJECTIVE: Human papillomaviruses, herpes simplex viruses, and Chlamyd ia trachomatis are very common infections of the genital tract. The pu rpose of our study was to develop a polymerase chain reaction-based as say for the simultaneous detection of these organisms from a single ge nital swab. STUDY DESIGN: To prove the technical feasibility of a simu ltaneous polymerase chain reaction assay for these organisms, a mixtur e of deoxyribonucleic acids extracted from cells infected by these thr ee agents was amplified in the same tube with three different sets of primers corresponding to specific regions of the human papillomavirus genome, the herpes simplex virus 1 and 2 genomes, and the Chlamydia tr achomatis plasmid, respectively. Then genital swabs from patients with suspected infection by one or more of these agents were assayed by po lymerase chain reaction for the presence of herpes simplex virus, huma n papillomavirus, and Chlamydia trachomatis independently and simultan eously. Most of the samples were analyzed in parallel by other methods : herpes simplex virus by culture, Chlamydia trachomatis by culture an d antigen staining, and human papillomavirus by the filter in situ hyb ridization method. RESULTS: Analysis of the polymerase chain reaction products amplified from the deoxyribonucleic acid mixture revealed thr ee bands corresponding to the respective amplified region of each micr oorganism. A total of 391 genital swabs were assayed independently by polymerase chain reaction for the presence of herpes simplex virus (11 3 samples), human papillomavirus (200 samples), and Chlamydia trachoma tis (78 genital swabs and four urethral swabs). Forty-nine were herpes simplex virus positive (47 by culture), 45 were human papillomavirus positive (43 by filter in situ hybridization), and one sample was posi tive for Chlamydia trachomatis, both by polymerase chain reaction and by culture. Ninety-two of the 391 samples were analyzed simultaneously by polymerase chain reaction for the presence of the three agents. Th e correlation between the results obtained independently and simultane ously was of the order of 100%: 29 were positive for herpes simplex vi rus, 16 were positive for human papillomavirus, and one was positive f or Chlamydia trachomatis. In one sample we could detect both human pap illomavirus and herpes simplex virus. CONCLUSIONS: The polymerase chai n reaction simultaneous assay is a quick and efficient way of detectin g herpes simplex virus, human papillomavirus, and Chlamydia trachomati s from a single genital swab. This method can greatly simplify the dia gnostic procedures in the laboratory.