title: Comparison of multiplex PCR assay with culture for detection of genital mycoplasmas.
abstract: Ureaplasma, spp. Mycoplasma genitalium, and Mycoplasma hominis are associated with infection of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. We have developed a multiplex PCR for the detection of these Mycoplasmatales in a single amplification reaction. The analytical sensitivities of this assay were 10.8, 10.8, and 8.8 CFU for each organism, respectively. This multiplex PCR was compared to culture on 26 cervical swabs, 2 vaginal swabs, 4 female urine specimens, 49 semen samples, 2 male urine specimens, and 1 nonspecified sample. A total of 21 specimens were culture positive (25%); 17 of these were PCR positive. An additional 11 specimens were PCR positive but culture negative. Of the 21 culture-positive specimens, 17 (81%) grew Ureaplasma spp. and 4 (19%) grew Mycoplasma spp. Of the 28 PCR-positive specimens, Ureaplasma spp. was detected in 23 (82%), M. hominis was detected in 3 (11%), and both were detected in 2 (7%). In a confirmatory analysis, all samples were tested by amplification of a second target of the ureaplasma genome. True-positive cases were defined as a positive result by culture or by both amplification assays. The multiplex PCR detected organisms in 26 of the 30 true-positive specimens, as well as in 2 other specimens. Based on a 36% prevalence of infection, the sensitivity, specificity, and positive and negative predictive values of multiplex PCR analyses were 87, 96, 94, and 93%, respectively. Multiplex PCR offers a rapid, sensitive, and easy method to detect genital mycoplasmas.
authors.affiliation: Department of Pathology and Laboratory Medicine, Albany Medical Center, Albany, New York 12208, USA. stellrk@mail.amc.edu
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Stellrecht
authors.forename: Amy M
authors.initials: AM
authors.lastname: Woron
authors.forename: Nada G
authors.initials: NG
authors.lastname: Mishrik
authors.forename: Richard A
authors.initials: RA
authors.lastname: Venezia
mesh_terms: D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D005831:Genital Diseases, Female / Q000382:microbiology*; D005832:Genital Diseases, Male / Q000382:microbiology*; D006801:Humans; D008297:Male; D009178:Mycoplasmataceae / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D009180:Mycoplasmatales Infections / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial