Simultaneous detection of three common sexually transmitted agents by polymerase chain reaction

Human papillomaviruses, herpes simplex viruses, and Chlamydia trachomatis are very common infections of the genital tract. The purpose of our study was to develop a polymerase chain reaction-based assay for the simultaneous detection of these organisms from a single genital swab.

To prove the technical feasibility of a simultaneous polymerase chain reaction assay for these organisms, a mixture of deoxyribonucleic acids extracted from cells infected by these three 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 trachomatis plasmid, respectively. Then genital swabs from patients with suspected infection by one or more of these agents were assayed by polymerase chain reaction for the presence of herpes simplex virus, human papillomavirus, and Chlamydia trachomatis independently and simultaneously. Most of the samples were analyzed in parallel by other methods: herpes simplex virus by culture., Chlamydia trachomatis by culture and antigen staining, and human papillomavirus by the filter in situ hybridization method.

Analysis of the polymerase chain reaction products amplified from the deoxyribonucleic acid mixture revealed three bands corresponding to the respective amplified region of each microorganism. A total of 391 genital swabs were assayed independently by polymerase chain reaction for the presence of herpes simplex virus (113 samples), human papillomavirus (200 samples), and Chlamydia trachomatis (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 positive 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. The correlation between the results obtained independently and simultaneously was of the order of 100%: 29 were positive for herpes simplex virus, 16 were positive for human papillomavirus, and one was positive for Chlamydia trachomatis, in one sample we could detect both human papillomavirus and herpes simplex virus.

The polymerase chain reaction simultaneous assay is a quick and efficient way of detecting herpes simplex virus, human papillomavirus, and Chlamydia trachomatis from a single genital swab. This method can greatly simplify the diagnostic procedures in the laboratory.