Abstract: OBJECTIVE To develop a novel method based on polymerase chain reaction-ligase detection reaction(PCR-LDR) system for simultaneous detection of mutations in the gene locus of various major drug metabolizing enzymes in human. METHODS Single nucleotide polymorphism(SNP) sites related to drug metabolism enzymes were selected, and PCR primers and LDR probes at each SNP site were designed and synthesized. DNA extracted from human oral mucosa cells was used as template to obtain PCR-LDR reaction junction products. ABI 3130XL was used for detection and analysis. RESULTS The PCR-LDR typing method established in this study was used to classify 14 SNP sites from different individuals, and the typing results were completely consistent with the sequencing results. CONCLUSION The PCR-LDR typing method established in this study is able to perform one-time typing for 14 SNP sites at the same time with accurate and reliable results. It is a simple, effective and low-cost SNP typing method.