Abstract: 　Objective　To analyze the genotype and variation of Mycoplasma pneumonia (MP) strains isolated from children with MP infection in Soochow area. Methods　The nasopharyngeal secretions from hospitalized children with MP infection were collected during January 2012 and December 2013. The nested-multiplex PCR based on MP P1 gene was performed to detect the subtype of MP gene. Results　In 313 samples, 304 (97.12 %) samples were classified as P1-I type and 8 (2.56%) samples were classified as P1-II type and one (0.32%) was V2 variant. Gene sequencing results were consistent with nested-multiple PCR results. Conclusions　Nested-multiplex PCR is a reliable method for genotyping of MP P1 gene. During the study period, P1-I type was the common genotype and only one case of V2 variant was found.