[PCR-single-strand conformation (SSCP), DNA direct sequencing analysis in detecting mutation in exon 2 of g6pd gene]

Hereditary glucose-6-phosphate dehydrogenase (G6PD) in red blood cell was one of the most common genetic diseases in South China. The research data of G6PD gene showed that at least 6 point mutations were responsible for various G6PD variants in Chinese. For developing a rapid, sensitive, and effective method to detect point mutation, we applied single-strand conformation (SSCP) analysis for detection of mutation in exon 2 of G6PD gene of 20 cases of G6PD variants. Four of them were found that mobility shift band in one of two single strands DNA is slower than other individuals. PCR direct sequencing for these 4 samples were given a base substitution (T to D) at nucleotide 95 of cDNA. The results indicated that this technique is very simple, sensitive, and useful over other methods of detecting point mutation. The experimental conditions of PCR-SSCP and features of this mutation were discussed at the same time.