In this retrospective study we employed real-time polymerase chain reaction (PCR) to analyse the occurrence of Mycoplasma pneumoniae among upper and lower respiratory tract infections (RTI) in the Central Region of Slovenia between January 2006 and December 2014. We also used a culture and pyrosequencing approach to genotype strains and infer their potential macrolide resistance. Of a total 9,431 tested samples from in- and out-patient with RTI, 1,255 (13%) were found to be positive by M. pneumoniae PCR. The proportion of positive samples was 19% (947/5,092) among children (?16 years-old) and 7% (308/4,339) among adults (>16 years-old). Overall, among those PCR tested, the highest proportions of M. pneumoniae infections during the study period were observed in 2010 and 2014. In these two years, 18% (218/1,237) and 25% (721/2,844) of samples were positive respectively, indicating epidemic periods. From the 1,255 M. pneumoniae PCR-positive samples, 783 (614 from paediatric and 169 from adult patients) were successfully cultured. Of these, 40% (312/783) were constituted of strains belonging to the P1 type II genomic group, while 60% (469/783) contained strains of the P1 type I group. Two isolates comprised both P1 type I and II strains. Results of a genotype analysis by year, showed that the dominant M. pneumoniae P1 type during the 2010 epidemic was P1 type II (82% of isolates; 81/99), which was replaced by P1 type I in the 2014 epidemic (75%; 384/510). This observation could indicate that the two epidemics may have been driven by a type shift phenomenon, although both types remained present in the studied population during the assessed period of time. Only 1% of strains (7/783) were found to harbour an A2063G mutation in the 23S rRNA gene, which confers macrolide resistance, suggesting that the occurrence of M. pneumoniae macrolide resistance still seems to be sporadic in our geographic area.