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To understand whether there were mixed infection and the existence feature of PE_PGRS33 gene subpopulations individual in pediatric tuberculosis patients.One hundred and sixteen clonal isolates were obtained from the L-J culture of their sputum/gastric aspirate from 5 pediatric tuberculosis patients and cerebrospinal fluid and there were 3~16 isolates for each patient.All of the clonal isolates were subjected to genotyping with the variable-number tandem-repeat (VNTR) system with 24 loci used PCR and DNA sequencing to identify genetic variations in the PE_PGRS33 gene in comparison with H37Rv.One hundred and sixteen M.tuberculosis clonal isolates were identified five genotypes and two types sequenced variations of PE_PGRS33 gene.There was no difference in genotype and sequence variations of PE_PGRS33 gene among the clonal isolates from different parts from a same child, and it implied that each of 5 pediatric patients with tuberculosis had the infection with a single M.tuberculosis strain and the existence feature of PE_PGRS33 gene subpopulations was genetic homogeneity.