Research Of The Three G6PD Gene Mutations In Neonate From Chong Qing City

Objective: Heredity glucose-6-phosphate dehydrogenase(G6PD) deficiency in red blood cells is the most common enzymopathy of humans, affecting approximate 400 million people worldwilde. there is a regional high frequency in southern China, there is a regional high frequency in ChongQing. It is estimated that the incidence of disease of G6PD is 4%-15%.individual area reachs 40%. the G6PD gene is "housekeeping"gene typically, and is located on region Xq28,roughly 20114 bp,13 exons,and 12 introns. coding spacer long is 1548bp,and codes 515 amino acids. According to the suggested World Health Organization(WHO) Criteria of identify of G6PD biochemical variant types, namely indes of enzymatic activity,electrophoresis velocity, substrate Michaelis constant, NADP appetency, fingerprint analysis and so on. so far more than 400 different G6PD biochemical variant types heve been identified, and 20 of 400 biochemical variant types can cause hemolysis.The biochemical type of normal G-6PD is B- type, there is variant type-A of a sort of enzyme alive reduce among africa crowd. so far, Our country have discorved at least more than 40 kinds of B-G6PD variant typesof HongKong ,Guangdong, HaKKa,White sand of miao and Taiwan, and so on.Currently, 126 different types of gene mutations are identified in the world, in addition to 5 species deletion, and the rest is all G6PD gene point mutations.Therearel5differenttypesofgeneMutationsinourcontry.TheyareG138 8A,G1376T,A95G,G1388A,G1376T,A95G,C1311T,G392T,C1024T,C592T,Cl 004T,A493G,G487A,C1360T,A835T,G1381A,G1387T,G871A.Biochemical variant types bear no relation to point mutation types, various biochemical variants share the same mutation, a few of point mutations share the same biochemical variant. G6PD gene point mutations result in G6PD deficiency clinical manifestations. For making a diagnosis and giving treatment neonatal patients with G6PD deficiency accurately as soon as possible, lowering the death rate, and improving survival quality , During twenty-one month from 2001-12-01 to 2003-09-30, we researched the mutation types of three G6PD genes in 54 case neonatal patient with G6PD deficiency from Chong Qing City, and estimated the approximate frequencies of gene mutations and discussed the clinical significance and genetics characteristic.Methods: Totally G6PD three gene mutations 54 out of 78 cases who were diagnosed neonate patients with G6PD deficiency testified by quantitative analysis of nitroblne terazolium or by G6PD gene determination were tested by PCR instrument.Diagnosis all accords with Zhangzhi-Lan' criterion of blood illness diagnosis and effect, two milliliter of heparin demurcruor was taken out. white blood cells and DNA were abstracted by normal method.Experiments were designed positive, negative and internal contrast,and were separately added to G1388A,G1376T and A95G positive samples, and normal DNA samples,and Mediterrean anemia primer that expanded 601bp gene fragment between transcribed promoter spacer of 6-bead albumen gene and the second exon. PCR circle expand was carried through by amplification refractomutation system set upl388M,1376Mand95M, DNA product was observed special band by dye and electrophoresis under violet exo lamp.Results : 54 cases of neonatal patients with G6PD deficiency were examined, and 39 of them were identified to be G1388A(72%), 8 were G1376T(15%), The G1388A and G1376T mutations account For 87% of the 54 cases and were the main types of G6PD gene mutations of infant, newborn in Chong Qing .The rest(13%) belong to undefined mutations.Conclusions: G1388A and G1376T gene mutations were described for the first time in infant, newborn in Chong Qing. Research had proved that AMRS methods could be used in detecting common mutations of G6PD gene with the benefits of simple, fast, economic, and accurate,but that AMRS method was of great benefits to clinical diagnosis, population genetics, anthropology and gene geography. Detection of G6PD gene mutations was served as a kind of genetics index of our people origin and migration and...