Application of Mass-array gene chip to detect glucose-6-phosphate dehydrogenase gene mutations

Abstract

Objective To develop the Mass-array gene chip to detect glucose-6-phosphate dehydrogenase (G6PD) gene mutations, and to evaluate its quality. Methods Randomly choosing the children who perform neonatal screening in Neonatal Screening Center of Fujian Maternity and Children Health Hospital from 2006 to 2013. Children were divided into control group and G6PD patient group. Using Genotyping Tools from Sequenom company and the software of Mass-array Assay Design to design the PCR amplification primer of 33 G6PD gene mutations which were well-known in Chinese. Then depending on Mass-array gene chip technology to detect glucose-6-phosphate dehydrogenase gene mutations. DNA Sanger sequencing was used to verify the accuracy of the testing results of gene chip. Results In the 300 children with G6PD deficiency, we detected 9 kinds of simplex single point mutations: 1376G>T, 1388G>A, 95A>G, 1024C>T, 392G>T, 1360C>T, 487G>A, 517T>C, 1365-13T>C and 7 kinds of compound mutations: 871G>A/1365-13T>C/1311C>T, 1004C>A/1311C>T/1365-13T>C, 1376G>T/ 1365-13T>C/1311C>T, 1365-13T>C/1311C>T, 1376G>T/1365-13T>C, 95A>G/1365-13T >C/1311C>T, 1388G>A/1365-13T >C. No G6PD gene mutation was detected in 300 normal children. The results of DNA Sanger sequencing were identical to that of gene chip. Conclusion Mass-array of G6PD gene is an effective, accurate and efficient way for G6PD gene mutation screening.(Chin J Lab Med, 2015, 38: 822-826) Key words: Glucosephosphate dehydrogenase; Mutation; Oligonucleotide array sequence analysis; Sequence analysis, DNA; Polymerase chain reaction