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Journal of Clinical Microbiology, April 2004, p. 1528-1533, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1528-1533.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Comparison of Multiplex PCR Assay with Culture for Detection of Genital Mycoplasmas

Kathleen A. Stellrecht,* Amy M. Woron,{dagger} Nada G. Mishrik,{ddagger} and Richard A. Venezia§

Department of Pathology and Laboratory Medicine, Albany Medical Center, Albany, New York 12208

Received 9 May 2003/ Returned for modification 24 June 2003/ Accepted 4 December 2003

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.

* Corresponding author. Mailing address: Department of Pathology and Laboratory Medicine, Division of Clinical Microbiology, MC-22, Albany Medical Center, 43 New Scotland Ave., Albany, NY 12208. Phone: (518) 262-3587. Fax: (518) 262-4337. E-mail: stellrk{at}mail.amc.edu.

{dagger} Present address: Wadsworth Center, New York State Department of Health, Albany, N.Y.

{ddagger} Present address: National Center for Infectious Diseases, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Ga.

§ Present address: Laboratories of Pathology, School of Medicine, University of Maryland, Baltimore, Md.

Journal of Clinical Microbiology, April 2004, p. 1528-1533, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1528-1533.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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