Gene Analysis Of Glucose-6-phosphate Dehydrogenase(G6PD) In Neonates Of Guangdong

Glucose-6-phosphate dehydrogenase(G6PD) is a metabolic disorder induces hemolysis.The cause of the disease is G6PD gene mutation.It's an X-link incomplete dominant inheritance disease.The incidence of hyperbilirubinemia in G6PD deficiency neonates is higher than the normal neonates.One of the most important complication of G6PD deficiency is severe neonatal hyperbilirubinemia and the risk of developing kerniterus,which may cause the permanent nervous system damages even the death.So it's very important to diagnose and treat as early as possible. G6PD deficiency is the most common single gene inheritance disease in human estimated to affect 400 million individuals worldwide.The incidence of G6PD deficiency in Guangdong and Guangxi province is higher than the other provinces and regions in China.There are at least 25 different point mutations associated with reduced G6PD enzyme activity have been identified in the Chinese G6PD gene till now.The three commonest mutations in Chinese are G6PD Canton(1376G>T),G6PD Kaiping(1388G>A) and G6PD Gaohe(95A>G).Molecular analysis to investigate the G6PD gene mutation helps to diagnose at genic level and value the therapeutic effect.Objective:We have analysed 50 Cantonese G6PD deficiency neonates and 20 normal neonates with PCR-based techniques to identify their genotypes.To investigate the genotypic frequency of G6PD Canton(1376G>T) ,G6PD Kaiping(1388G>A),G6PD1311C>T and G6PD IVS93 T>C.That helps to get the message of the character and the pathogenesis of G6PD gene mutation.Methods:To extract genomic DNA by standard method from all 50 G6PD deficiency Cantonese subjects and 20 neonates of control group.We have identified G6PD Canton(1376G>T) , G6PD1311C>T and G6PD IVS93 T>C by using PCR-direct DNA sequence analysis and identified G6PD Kaiping(1388G>A) by using Allele specific amplification system (ARMS).Data of different genotypes has been analysed using the SPPS software.Results:The overall results of mutation analysis in the 50 G6PD deficiency subjects showed the existence of 4 different alleles: G6PD Canton(1376G>T), G6PD Kaiping(1388G>A), cnDNA1311C>T,11 Intron 93T>C.The different genotypic frequency is: G6PD Kaiping(1388G>A) 48.0%(24/50), Canton(1376G>T) 12.0%(6/50), 1388G>A/1376G>T 12.0%(6/50), 1388G>A/1311C>T/IVS 93T>C 2.0%(1/50),1376G>T/1311C>T/IVS 93T>C 2.0%(1/50).We have identified a G6PD1388G>A mutation in the control group. There was significant differences in the Male/Female, mean peak serum bilirubin, MetHb% and G6PD/6PGD between the 3 mutation groups (1388G>A,1376G>T,1388G>A/1376G>T) and the control group. There was no significant differences in the Male/Female, mean peak serum bilirubin, MetHb % ,G6PD/6PGD between the 3 mutation groups. There was no significant difference in the onset of jaudince between the 3 mutation groups and the control group ,and between the 3 mutation groups neither. There was significant difference in the finale of jaudince between the 1376G>T group,1388G>A/1376G>T group and the control group. There was significant difference in the finale of jaudince between the 1388G>A group and 1388G>A/1376G>T group. There was no significant difference in the finale of jaudince between the 1388G>A group and the control group, 1376G>T group and 1388G>A group , 1376G>T group and 1388G>A/1376G>T group. There was significant difference in the hemoglobin between the 1388G>A group and the control group, 1376G>T group and 1388G>A group. There was no significant difference in the hemoglobin between the 1376G>T group and the control group,1388G>A/1376G>T group and the control group. There was no significant difference in the hemoglobin between the 1376G>T group,1388G>A group and 1388G>A/1376G>T group. There was no significant differences in the duration of phototherapy and albumin between the 3 mutation groups (1388G>A,1376G>T,1388G>A/1376G>T). Conclusion:1. G6PD Kaiping(1388G>A) and G6PD Canton(1376G>T) are the two commonest mutation in Cantonese.The genotypic frequence of G6PD 1388G>A is 60.0%. The genotypic frequence of G6PD 1376G>T is 26.0%.2. G6PD gene complex mutation has been confirmed by this study.We have detected the 1388G>A/1376G>T, 1388G>A/1311C>T/IVS 93T>C and 1376G>T/1311C>T/IVS 93T>C complex mutation of G6PD gene.3. The female heterozygote of G6PD deficiency is likely to have variable enzyme activities.The result of the G6PD/6PGD is likely to at the normal range in the female heterozygote of G6PD deficiency.4. G6PD deficiency is one of the high risk factors of neonatal hyperbilirubinemia,but it's not the high risk factor to make the neonatal jaundice earlier than the normal infant.We found no significant difference in the neonatal jaundice between the solo mutation group and the complex mutation group.