Abstract: Objective: To analyze the gene mutations of two pedigrees with distal renal tubular acidosis. Methods: The data of children with distal renal tubular acidosis were collected, and their mutations of genomic DNA were screened by the high throughput sequencing technology. The suspected mutations were verified by Sanger sequencing, and then bioinformatics prediction was performed. Results: The sequencing results showed that a heterozygous c.1185delC and c.1418C＞T mutation of ATP6V0A4 gene existed in one pediatric patient and that a heterozygous c.639+1G＞A and c.2227C＞T mutation of ATP6V0A4 gene existed in the other. The mutations of the two pediatric patients were inherited from their parents, respectively. The mutations of c.1418C＞T and c.2227C＞T had not been reported yet, and they were predicted to be harmful by the bioinformatics analysis. According to the standards and guidelines of the American College of Medical Genetics (ACMG), the two mutations were classified as likely pathogenic variation. Conclusion: Two new candidate mutation sites of ATP6V0A4 gene are found, which may be used for gene detection and genetic diagnosis and treatment.