SIMULTANEOUS PCR DETECTION OF HAEMOPHILUS-DUCREYI, TREPONEMA-PALLIDUM, AND HERPES-SIMPLEX VIRUS TYPE-1 AND TYPE-2 FROM GENITAL ULCERS

A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus duc reyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2 . By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three in tervals from 1992 through 1994 were evaluated. The results of the M-PC R assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results w ere available for 99 specimens collected during the third interval. Co nfirmatory PCR assays targeting different gene sequences for each of t he three organisms were used to validate the M-PCR results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M -PCR and the confirmatory PCR were positive. The resolved sensitivitie s of M-PCR for HSV, H. ducreyi, and T. pallidum were 100, 98.4, and 91 %, respectively. The resolved sensitivities of HSV culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81%, respecti vely. These results indicate that the M-PCR assay is more sensitive th an standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.