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Background  In China mainland, most  Mycoplasma pneumoniae  related studies are carried out in Beijing and Shanghai, while rare studies are performed in the other regions. In this study, we analyzed the molecular biology characteristics and antimicrobial susceptibility of clinical isolates of  M. pneumoniae  from 5 regions between January 2017 and December 2018.    Methods  Genotyping was performed to 154  M. pneumoniae  isolates from 5 cities using PCR and multiple-locus variable-number tandem repeat analysis (MLVA) method. Antimicrobial susceptibility test was performed to all the isolates against 4 antibiotics. Sequencing was performed to the amplification products of the 23S rRNA drug resistant gene.    Results  Genotype I was detected in 118  M. pneumoniae  isolates (76.6%), and genotype II was identified in 36 isolates (23.4%). The majority (92.2%) of the MLVA genotypes were 4–5–7-2 and 3–5–6-2, which represented the genotype I and II, respectively. The total macrolide (ML) resistance rate was 79.7%. The minimum inhibitory concentration (MIC) of the erythromycin was in a range of 128- > 256 μg/ml, while that for the azithromycin was 2-32 μg/ml. There were mutations in the 23S rRNA in each ML resistance isolate. Jilin city showed the highest prevalence of genotype I (100%) and ML resistance rate (100%), while Jinan showed the lowest prevalence of genotype I (45.5%) and ML resistance rate (54.5%).    Conclusions  A large variance was identified in the  M. pneumoniae  genotype and ML resistance among the 5 cities. The proportion of  M. pneumoniae  with a genotype II genotype (3–5–6-2) showed an increased trend.    Electronic supplementary material  The online version of this article (10.1186/s13756-019-0576-5) contains supplementary material, which is available to authorized users.

Antimicrobial susceptibility and molecular characteristics of Mycoplasma pneumoniae isolates across different regions of China