Implementation of a quantitative-polymerase chain reaction for the detection of Mycoplasma genitalium
Abstract
The diagnosis of M. genitalium infections by bacteriological culture is not feasible due to slow growth and that is time-consuming. Consequently, molecular methods using DNA amplification are widely used in the infection diagnosis procedure. There are no reports in Cuba of the implementation and use of quantitative Polymerase Chain Reaction methods for the detection and quantification of this pathogen. The aim of this study was to implement a qPCR method for the detection of M. genitalium by the amplification of the mgpBadhesin gene using two LightCycler® (Roche) protocols, SYBR Green I and TaqMan. The specifity and sensitivity were evaluated by using DNA of M. genitalium as well as other mycoplasms of human origin. In both qPCR-protocols, a limit of detection of 3.6 genome equivalents per µL template (geq/µL) was reached. The TaqMan protocol showed better efficiency than the SYBR Green assay. Both protocols showed a high specificity for the detection of M. genitalium, without cross-reactions with other mycoplasmas of human origin. For the first time in Cuba, a qPCR for detection of M. genitalium was implemented. The TaqMan method showed a better performance than the SYBR method and should be used for future applications in clinical samples. The present work will allow performing future studies of genetic and antigenic characterization of the circulating strains in Cuba, useful as vaccine immunogen.References
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