The Spread of Mycoplasma pneumoniae Is Polyclonal in Both an Endemic Setting in France and in an Epidemic Setting in Israel

Arabella Touati,Allon E Moses,Cécile Bébéar,Mitchell F Balish,Ran Nir-Paz,Sabine Pereyre,Carlos Hidalgo-Grass,Alain Charron

doi:10.1371/journal.pone.0038585

Abstract

Mycoplasma pneumoniae infections occur both endemically and epidemically, and macrolide resistance has been spreading for 10 years worldwide. A substantial increased incidence of M. pneumoniae infections has been reported in several countries since 2010. Whether this increased incidence is attributed to different or to the same M. pneumoniae genotype is unknown. We have developed a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) for the molecular typing of M. pneumoniae isolates. In this study, the MLVA typing method was modified and validated to be applicable directly to respiratory tract specimens without culture. This method was applied to 34 M. pneumoniae-positive specimens received at the Bordeaux Hospital, France, between 2007 and 2010 in an endemic setting, and to 63 M. pneumoniae-positive specimens collected during an epidemic surge of M. pneumoniae infections in 2010 in Jerusalem, Israel. The M. pneumoniae endemic spread was shown to be polyclonal in France, with 15 MLVA types identified. Strikingly, the Israeli epidemic surge was also a multi-clonal phenomenon, with 18 circulating MLVA types. The macrolide resistance-associated substitution, A2058G, was found in 22% of the Israeli patients. Macrolide-resistant M. pneumoniae belonged to four MLVA types, the MLVA type Z being the most frequent one. An association between the MLVA type Z and macrolide resistance might exist since macrolide resistance was present or generated during the course of illness in all patients infected with this MLVA type. In conclusion, the discriminatory power of the MLVA showed that the spread of M. pneumoniae strains in France in an endemic setting was polyclonal as well as the surge of M. pneumoniae infections in Israel in 2010.

Highlights

Mycoplasma pneumoniae infections occur both endemically and epidemically worldwide, especially in children and young adults [1]
Several typing methods based on this gene were developed including PCR-restriction fragment length polymorphism (RFLP) [7,8], amplification and gene sequencing [9], real-time PCR with high-resolution melt analysis [10] and pyrosequencing [11]
We developed a multi-locus variable-number tandem-repeat (VNTR) analysis (MLVA) for the molecular typing of M

Summary

Introduction

Mycoplasma pneumoniae infections occur both endemically and epidemically worldwide, especially in children and young adults [1]. A substantial increased incidence was reported since 2010 in several countries of Northern Europe [2,3,4] and Israel [5,6]. Whether this increased incidence is attributed to different or to the same M. pneumoniae genotype is unknown. Until recently, the most common typing methods of M. pneumoniae were based on the analysis of the gene encoding the P1 protein. Several typing methods based on this gene were developed including PCR-restriction fragment length polymorphism (RFLP) [7,8], amplification and gene sequencing [9], real-time PCR with high-resolution melt analysis [10] and pyrosequencing [11]. Isolates were poorly differentiated due to the separation of isolates into only two types and a few variants related to each type [12,13,14,15]

Methods

Results

Conclusion