CAJ | 학술논문

目的建立变性高效液相色谱筛查G6PD基因变异的技术.观察广东汉族和壮族G6PD基因变异型的分布.方法应用变性高效液相色谱技术筛查广东汉族和壮族G6PD基因变异型.与ARMS及DNA测序比较,考察其准确性、灵敏度、效率及耗资.结果在73例G6PD缺陷者中发现G6PD 1388 G→A20例、1376 G→T24例、95 A→G 6例、1311 C→T 8例、1024 C→T 3例、392 G→T 2例、871 G→A 1例、1376 G→A/95 A→G4例、1388 G→A/95 A→G 3例、1311 C→T/95 A→G 2例,与DNA测序结果吻合;在酶学正常对照组中发现男1例女2例1388突变.结论变性高效液相色谱技术对G6PD基因变异型的筛查具有准确、高效、半自动化及经济等优点.尤其对无症状女性杂合子及临床Ⅳ型个体的筛查具有重要意义.
목적건립변성고효액상색보사사G6PD기인변이적기술.관찰엄동한족화장족G6PD기인변이형적분포.방법 응용변성고효액상색보기술사사엄동한족화장족G6PD기인변이형.여ARMS급DNA측서비교,고찰기준학성、령민도、효솔급모자.결과재73례G6PD결함자중발현G6PD 1388 G→A20례、1376 G→T24례、95 A→G 6례、1311 C→T 8례、1024 C→T 3례、392 G→T 2례、871 G→A 1례、1376 G→A/95 A→G4례、1388 G→A/95 A→G 3례、1311 C→T/95 A→G 2례,여DNA측서결과문합;재매학정상대조조중발현남1례녀2례1388돌변.결론변성고효액상색보기술대G6PD기인변이형적사사구유준학、고효、반자동화급경제등우점.우기대무증상녀성잡합자급림상Ⅳ형개체적사사구유중요의의.
Objective Of denaturing high performance liquid chromatography (DHPLC), a technique platform was developed for screening G6PD deficient variants. Methods When applied to screen and identify the G6PD deficient variants from 124 patients who come from 11 nations in China,the DHPLC was compared with amplification refractory mutation system (ARMS) or DNA sequence technique;and assessed carefully in its accuracy, sensitivity; efficiency and the cost of experiment. Results The G6PD-deficient variants, such as 1388 G→A(36/124 cases), 1376 G→T( 35 ), 95A→G(14), 1024 C→T (3),392 G→T(4), 871 G→A/1311 C→T/IVS XI +93 t→c(9) , 871 G→A (1), 1311C→T/IVS XI + 93 t→c (4),1376 G→T/1388 G→A (1) and so on, were characterized as sharp peaks by DHPLCand verified by DNA sequence. Further, the standard chromatograms were put into database for 8 kinds of common G6PDdeficient variants in Chinese populations. And also DHPLC found 3 G6PD variants (1388 G→A) from 103 negative controls. With taking 8.8 minutes and costing 1 dollar for each sample,DHPLCsuccessfully detected and identified 34 heterozygous females from patients with G6PD deficiency. However, ARMS checked 83 positive controls but got 12 false G6PD mutants, of which 5 were false positive, 7 false negative. Above results show that DHPLC sounds like to be more convenience, sensitive and accurate than ARMS and DNA sequence techniques for checking G6PD mutants. Conclusion DHPLC is of great advantage to screen the G6PD deficient variants with accuracy, convenience, automation andless cost,and significantly to identify the female heterozygote and clinical type Ⅳ individuals with G6PD deficiency.