Quantitative Detection of Mycoplasma genitalium from First-Pass Urine of Men with Urethritis and Asymptomatic Men by Real-Time PCR

Abstract

ABSTRACT We developed a TaqMan-based real-time PCR assay for quantifying Mycoplasma genitalium . This assay is able to specifically quantify concentrations of the M. genitalium 16S rRNA gene ranging from 10 7 to 10 copies/reaction. Using the TaqMan assay, we quantified the M. genitalium 16S rRNA gene in first-pass urine of men with urethritis and asymptomatic men who were positive for M. genitalium by PCR- and phylogeny-based assay. Of 130 men with gonococcal urethritis (GU), five were positive for M. genitalium . The mycoplasma load for each specimen was <5 × 10 copies/ml. Of 84 men with chlamydial non-GU (CNGU), seven were positive for M. genitalium . One man had an M. genitalium load of <5 × 10 copies/ml, and six men had loads ranging from 1.1 × 10 7 to 2.7 × 10 2 copies/ml. Of 86 men with nonchlamydial NGU (NCNGU), 17 were positive for M. genitalium . The mycoplasma loads for these men ranged from 3.3 × 10 6 to 2.3 × 10 2 copies/ml. Of 76 asymptomatic men, only two were positive for M. genitalium . For these men, the loads were 2 × 10 2 and <5 × 10 copies/ml. The patients with NGU had significantly higher concentrations of M. genitalium in their first-pass urine than did men with GU ( P < 0.01) or asymptomatic men ( P < 0.05). In addition, M. genitalium loads were significantly higher in men with NCNGU than those in asymptomatic men ( P < 0.05). The quantitative assessment of M. genitalium loads by the TaqMan assay will provide useful information for understanding the pathogenicity of this mycoplasma in the urogenital tract.