Comparison of the new Mycofast Revolution assay with a molecular assay for the detection of genital mycoplasmas from clinical specimens

Abstract

BackgroundGenital mycoplasmas are opportunistic bacteria that are associated with undesirable gynaecologic and reproductive events. Mycoplasmas are fastidious bacteria with increasing resistance to routine antimicrobials and often fail to grow on conventional culture methods. The commercial Mycofast Revolution assay permits the phenotypic detection and identification of genital mycoplasmas. Antimicrobial susceptibility testing against five antimicrobial agents with MICs corresponding to the CLSI guidelines can also be performed. This study aimed to compare the new commercially available Mycofast Revolution assay with a multiplex PCR assay.MethodsSelf-collected swabs were obtained from pregnant women attending the antenatal clinic of a tertiary academic hospital in Pretoria, South Africa from October 2012 to November 2012. These swabs were used to seed UMMt and modified Amies transport media. The seeded UMMt transported medium was used to inoculate the Mycofast Revolution assay for the identification, enumeration and antimicrobial susceptibility testing of genital mycoplasmas. Following DNA extraction from the modified Amies transport medium, specimens were subjected to a multiplex PCR assay for the detection of genital mycoplasmas.ResultsThe Mycofast Revolution kit had a sensitivity and specificity of 77.3% (95% CI: 62.15% to 88.51%) and 80% (95% CI: 28.81% to 96.70%), respectively, against the PCR assay. The positive and negative predictive values were 97.1% (95% CI: 85.03% to 99.52%) and 28.6% (95% CI: 8.57% to 58.08%). Genital mycoplasmas were detected in 71.4% (35/49) of samples with the Mycofast Revolution assay with 49% (24/49) being Ureaplasma spp. and 22.4% (11/49) mixed strains. The multiplex PCR assay had a positivity rate of 89.8% (44/49) for genital mycoplasmas; mixed strains were present in 51% (25/49) of samples, Ureaplasma spp. in 16.3% (8/49) and M. hominis in 22.4% (11/49) of samples.ConclusionsThere was a fair agreement (κ = 0.319) between the Mycofast Revolution assay and the mPCR assay. With the high prevalence rates of genital mycoplasmas, fast and efficient diagnostic methods are imperative to treat infections and minimise complications. The Mycofast Revolution assay is simple to use, has a short turn-around time and interpretation of results are straightforward. This assay circumvents common problems experienced with conventional culture and molecular methods in diagnostic laboratories where skilled personnel are limited and can be used as an alternative diagnostic assay.

Comparison of Mycoplasma IES, Mycofast Revolution and Mycoplasma IST2 to detect genital mycoplasmas in clinical samples

“…For Mycofast Revolution testing, 300 μl of the seeded saline suspension was dispensed on UMMt (Ureaplasma Mycoplasma medium transport) medium (EliTech Diagnostic, Puteaux, France); 100 μl were dispensed into wells 1 to 20 of the tray, as previously described [11]. Fifty μl of Mueller Hinton supplement (S. Mh) (EliTech Diagnostic, Puteaux, France) were dispensed into wells 6 and 7.…”

“…Especially in low to moderate test volume laboratories it is still considered the most cost effective strategy. Nucleic acid amplification-based tests have higher specificity and sensitivity than culture and are the only means for detection of Mycoplasma genitalium and for discriminating between U.urealyticum and U. parvum [10][11][12]. They can be also completed in few hours, despite cost of equipment and reagents is significant and molecular diagnosis-trained personnel is required.…”

“…Nonetheless, culture is still performed to monitor the antimicrobial susceptibility profiles of these pathogens [8,13]. The development of commercially available diagnostic assays, which are based on liquid broth cultures, offer a simpler alternative to conventional culture [11,14]. Mycofast Revolution (EliTech Diagnostic, Puteaux, France) and MycoplasmaIST2 (BioMérieux, Marcy l'Etoile, France) are two widely used assays for the detection of M. hominis and U. urealyticum, similar with regards to identification, antimicrobial susceptibility testing and turnaround time.…”

Susceptibility and Antimicrobial Resistance of Genital Ureaplasma Parvum

“…azithromycin (AZM) and clarithromycin (CLR)) allow better tolerability and the once daily dosing benefit can increase compliance. Treatment with AZM was equally successful, compared to ERY but with fewer side effects [2]. Ureaplasma species (Ureaplasma spp.)…”

The Prevalence and Antibiotic Resistance Profiles of Mycoplasma hominis and Ureaplasma urealyticum at a University Hospital

“…U.urealyticum ve M.hominis tespitinde kısa zamanda sonuç vermesi nedeniyle tercih edilebilecek bir yöntem olmasına rağmen antimikrobiyal duyarlılık ile ilgili bilgi vermemesi ve maliyetinin yüksek olması bu yöntemin dezavantajıdır (4,13) . Ayrıca ürogenital örnekler-den U.urealyticum ve M.hominis tespitinde özel kit şeklinde içinde besiyeri ve antimikrobiyal duyarlılık testlerini de içeren sistemler kullanı-labilmektedir (3,12) . Çalışmamızda Mycoplasma IST 2 test kiti kullanılmıştır.…”