Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy, affecting more than 200 million people worldwide. To date more than 123 mutations in the G6PD gene have been discovered, among which 12 point mutations are found in the Chinese. Setting up a simple and accurate method for detecting these mutations is not only useful for diagnosing G6PD deficiency under some circumstances that it is difficult to measure the activity of the enzyme, but also for studying the frequency of the G6PD genotypes. The purpose of this study was to develop a simple, inexpensive and accurate method for detecting these common mutations. Microarraybased assay was described in this study. Samples from 198 G6PD-deficient persons were investigated. The DNA sequencing data supported the results obtained by microarraybased assay. Thus, we concluded that the microarray-based assay is a rapid, simple, inexpensive, and accurate method for detecting the most common G6PD gene mutations among the Chinese. This method involves the selective amplification of human G6PD gene with specific oligonucleotide primers, fragmentation and labeling of PCR products, followed by hybridization with allele-specific oligonucleotide (ASO) probes on chip.