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.

Frequency of sexually transmitted diseases and main methodological implications

“…Quantificare l'incidenza e la distribuzione di questi patogeni nella popolazione, mediante l'investimento in opportune strategie di controllo e prevenzione, è importante per ridurre la morbilità e la mortalità correlata a questo tipo di infezioni con ricadute, sicuramente, in termini di qualità di vita per i pazienti ma anche in termini di spesa sanitaria più complessa da calcolare. Numerosi sono i test di amplificazione degli acidi nucleici (NAATs) in Real-Time polymerase chain reaction (RT-PCR) per la rivelazione di Chlamydia trachomatis e Neisseria gonorrhoeae (11,23,32,45) mentre poco implementati quelli per la rilevazione di Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum e Ureaplasma parvum, nonostante numerosi studi abbiano dimostrato una maggiore sensibilità delle metodiche RT-PCR (1,2,28,41,44). Diversi sono i laboratori che stanno implementando la diagnostica molecolare e impiegano le metodiche colturali solo quando è necessario l'antibiogramma.…”

Frequency of Mycoplasma hominis and Ureaplasma urealyticum in Females With Urogenital Infections and Habitual Abortion History in Ahvaz, Iran; Using Multiplex PCR

“…Mycoplasmatales are associated with infection of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. Infection with genital mycoplasmas has been linked with infertility (2)(3)(4). Ureaplasma spp.…”

“…Ureaplasma spp. are the main cause of non-chlamydial, non-gonococcal urethritis and acute prostatitis (2). In pregnancy, Ureaplasma can cause chorioamnionitis and preterm delivery (5).…”

Banana Bunchy Top Viral Coat Protein (CP) Gene Expression Studies at Molecular Level in Hill Banana cv. Sirumalai (AAB)

“…Enzyme Linked Immunosorbant Assay (ELISA) is convenient and easy method but it has a limitation of being less sensitive when the viral load is low (Landgraf et al, 1991). In this context, Nucleic acid based polymerase chain reaction (PCR) (Edwards and Gibbs, 1994;Grieco and Gallitelli, 1999;Tettelin et al, 1999;Olmos et al, 2002;Stellrecht et al, 2004) and hybridization methods have the advantage of amplifying the target nucleic acid present even at very low level and it has become an attractive technique for the diagnosis of plant viral diseases (Mullis et al, 1986;Henson and French, 1993;Xie and Hu, 1994;McManus and Jones, 1995;Hafner et al, 1997a and b;Elnifro et al, 2000;Nassuth et al, 2000).…”