Detection of macrolide resistant Mycoplasma pneumoniae in England, September 2014 to September 2015

Abstract

Mycoplasma pneumoniae infection can cause pneumonia, particularly in children. Global increase in macrolide-resistant M. pneumoniae is of concern due to limited therapeutic options. We describe the detection of macrolide resistance-conferring mutations in 9.3% of 43 clinical specimens where M. pneumoniae was detected in England and Wales from September 2014-September 2015. This study aims to impact by highlighting the presence of macrolide resistance in M. pneumoniae positive patients, promoting increased clinical vigilance. Macrolide resistance determinationThe Bacteriology Reference Department, Public Health England (PHE), London, receives specimens from England and Wales for M. pneumoniae testing and confirmatory testing. Here we detected M. pneumoniae by qPCR in 60 clinical specimens from 60 patients (Cambridge, Leeds, London, Manchester, Nottingham and Oxford) that were submitted to PHE between 1 September 2014 and 1 September 2015. DNA extractions from specimens, where M. pneumoniae was detected, were screened for point mutations known to confer macrolide resistance. Mutations in domain V of the 23S rRNA were detected by a modified version of the method described by Li et al., 2009 [3], wherein the entire region of interest is amplified and sequenced as one product. Primers used were as follows: forward primer 5'-ATCTCTTGACTGTCTCGGC-3' and reverse primer 5'-TACAACTGGAGCATAAGAGGTG-3'.Of the 60 specimens, 17 (28.3%; 95% confidence interval (CI): 18.4--40.8) contained insufficient DNA to determine macrolide resistance-conferring mutations. Of the remaining 43 specimens mutations in the 23S rRNA known to confer macrolide resistance were found in four (9.3%; 95% CI: 3.1-22.2). Of these 43 specimens, 32 were from a single city in England, Leeds, and a single specimen among these was positive for the mutation, 3.1% (95% CI: 0.01-17.1). The cases identified with point mutations known to confer macrolide-resistant M. pneumoniae were in two women and two men, respectively, aged > 15 to <65 years old. Three were hospitalised with pneumonia (Table) with no known connection between patients.Interestingly, two of the macrolide-resistant cases were patients that had recently arrived from the United States (exact timeline unknown); of which one had received clarithromycin whilst undergoing treatment in the UK. The origins of the infecting M. pneumoniae strains in these two cases may have been external to England and Wales. The other two cases were from separate cities in England. All macrolide resistanceconferring mutations were A2058G (Escherichia coli numbering) point mutation in the 23S rRNA.

Mycoplasma pneumoniae infections, 11 countries in Europe and Israel, 2011 to 2016

“…Routine macrolide resistance monitoring was not systematically in place. This may contribute to the under-detection of resistance or reflect low levels of macrolide resistance reported across Europe [18][19][20]. High levels of resistance have been noted, for example, in Israel (30%) [21] and China wherein reported macrolide resistance levels are 90 to 100% [22,23].…”

Community-acquired pneumonia related to intracellular pathogens

“…Recent reports from Asia and some European countries describe the isolation of strains of M. pneumoniae resistant to macrolides [9,10].…”

“…Macrolides and quinolones remain the best empirical treatment of intracellular pathogens owing to their good antimicrobial activity and high intracellular concentrations. Although antibiotic resistance in these intracellular pathogens does not currently represent a clinical problem in the majority of settings, recent studies from Asia and some European countries report the isolation of strains of M. pneumoniae resistant to macrolides, suggesting a need to monitor these pathogens to evaluate the clinical impact of resistance in CAP [9,10].…”

Molecular detection and evaluation of resistance of Mycoplasma Pneumoniae to macrolide (Erythromycin) due to mutation in the 23S rRNA gene among Iranian Patients with respiratory infections

“…ML resistance is associated with a point mutation in domain V of 23S rRNA gene and ribosomal proteins L22 and L14. Mutation affecting 23S rRNA gene at A2063G position is the most common mutation, followed by A2064G position, which is mostly responsible for signi cant ML resistance in M. pneumoniae (10,23). Results of the analysis of the sequence of 23S rRNA gene showed that point mutations at A2431G and G2491A positions were reported only in one specimen.…”