Simultaneous PCR detection of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus types 1 and 2 from genital ulcers

Abstract

A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, 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 intervals from 1992 through 1994 were evaluated. The results of the M-PCR assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results were available for 99 specimens collected during the third interval. Confirmatory PCR assays targeting different gene sequences for each of the 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 sensitivities 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%, respectively. These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.

Genital ulcer disease and human immunodeficiency virus: a focus

“…51,55 Compared with M-PCR, culture had a sensitivity of 74%. 57 For lymphogranuloma venereum (LGV), microimmunofluorescence and PCR are of adequate sensitivity, while donovaniosis is best diagnosed by tissue smears. 54,55 Despite its sensitivity, nucleic acid amplification is not practical for diagnosing GUD.…”

Diagnosis and Monitoring of Infections

“…A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. The M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers (Orle et al, 1996).…”

Chancroid and human immunodeficiency virus infection – a review

“…Most ulcers appear to be chancroid on clinical grounds, although the H. ducreyi isolation rate is only 23%, perhaps reflecting increased antimicrobial use. 34 The use of PCR has revolutionized the laboratory diagnosis of chancroid, 35 because alternative methods of diagnosis, such as Gram's stain and culture, are inaccurate and insensitive as H. ducreyi is a fastidious organism. Several serologic tests have been developed, including adsorption enzyme immunoassay (EIA) and lipooligosaccharide (LOS) EIA, to detect H. ducreyi infection.…”