Detection of macrolide-resistance mutations in 23S rRNA gene of Mycoplasma pneumoniae using a novel real-time PCR assay

Abstract

Background. Mycoplasma pneumoniae is a widespread pathogen of the respiratory tract diseases. In recent years, the data of foreign publications indicate the appearance and spread of resistant M. pneumoniae to macrolide antibiotics, which is associated with mutations in the 23S pRNA gene, mainly in positions 2063, 2064 and 2617. Methods. Studied 146 clinical samples obtained in 2006-2013. of patients with infections of the lower respiratory tract. Primary screening was done for the presence of DNA material M. pneumonia based on polymerase chain reaction (PCR) in real time. Results. The possibility of detecting and differentiation of various mutations in positions 2063, 2064 and 2617 in the 23S pRNA gene format single sample multiplex PCR in real time with subsequent melting curve analysis of fluorescently-labeled probes. In the study of clinical samples in all cases been detected 23S pDNA sequences of M. pneumoniae “wild-type” typical for phenotype of sensitivity to macrolides. Conclusions. Detection of single nucleotide corresponding replacements allows effective predict macrolide resistance phenotype, but used for this purpose methods are laborious and expensive. At present data on the prevalence and mechanisms of macrolide resistance in strains of M. pneumoniae in Russia are absent. The present study focuses on the development and validation of a new method for the determination of mutations associated with resistance to mac-rolides in M. pneumoniae, and also its use for the analysis of clinical samples in patients with lower respiratory tract infections.