福建省龙岩市珠蛋白基因新位点突变的测序检测
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摘要
目的检测福建省龙岩市地中海贫血(地贫)患者可能携带的α珠蛋白基因和β珠蛋白基因新位点突变。方法选择福建省龙岩市地贫筛查阳性的15例患者,采用跨越裂口PCR法检测~(--SEA)、-α~(3.7)和-α~(4.2) 3种缺失型突变,PCR扩增α珠蛋白基因和β珠蛋白基因,Sanger测序检测纯化的PCR产物的序列,并且将感兴趣的标本用毛细管电泳技术检测血红蛋白的类型及占比。结果共检测到10种新位点突变,分别为HBA1:codon 14 TGG> TGA、HBA1:IVS-I-65A>G、HBA1:codon 139 AAA> ATA、HBA2:-43 G> C、HBA2:c.*136A> G、HBB:-99 G> A、HBB:-96 G> A、HBB:-62 G> A、HBB:c.56 C> T和HBB:3’UTR+133 G> C。15例地贫患者中有4例为HBA1:codon 14 TGG> TGA携带者。结论研究发现了10种珠蛋白基因的新位点突变,丰富了珠蛋白基因突变数据库。HBA1:codon 14 TGG>TGA在福建省龙岩市人群中可能是常见的α地贫位点突变,有必要进行大样本检测确定HBA1:codon 14 TGG>TGA在该地区地贫患者中的携带率。
Objective To detect the new point mutations of hemoglobin subunit alpha and beta genes probably carried by thalassemia patients from Longyan, Fujian Province in China. Methods Fifteen patients pos--itive for thalassemia from Longyan, Fujian Province were recruited in this study. Three deletion mutants of ~(--SEA),-α~(3.7) and-α~(4.2) were detected by the gap PCR. The hemoglobin subunit alpha and beta genes of thalassemia patients was amplified by PCR. The sequence of the purified PCR products was analyzed by Sanger sequencing. The hemoglobin type and proportion of interested samples were analyzed by capillary electrophoresis. Results Ten new point mutations of hemoglobin genes were found including HBA1: codon 14 TGG > TGA, HBA1: IVS-I-65 A > G, HBA1: codon 139 AAA > ATA, HBA2:-43 G > C, HBA2: c.*136 A >G, HBB:-99 G >A, HBB:-96 G > A, HBB:-62 G> A, HBB: c.56 C > T and HBB: 3'UTR +133 G > C. Four of the 15 thalassemia patients were HBA1: codon 14 TGG > TGA carriers. Conclusions Ten new point mutations of hemoglobin genes are detected, which enrich the hemoglobin gene mutation database. HBA1: codon 14 TGG>TGA may be a common alpha-thalassemia mutation in the population of Longyan, Fujian Province. Subsequent investigations with large sample size are urgently required to determine the carrying rate of HBA1: codon 14 TGG > TGA in thalassemia patients from this area.
Objective To detect the new point mutations of hemoglobin subunit alpha and beta genes probably carried by thalassemia patients from Longyan, Fujian Province in China. Methods Fifteen patients pos--itive for thalassemia from Longyan, Fujian Province were recruited in this study. Three deletion mutants of ~(--SEA),-α~(3.7) and-α~(4.2) were detected by the gap PCR. The hemoglobin subunit alpha and beta genes of thalassemia patients was amplified by PCR. The sequence of the purified PCR products was analyzed by Sanger sequencing. The hemoglobin type and proportion of interested samples were analyzed by capillary electrophoresis. Results Ten new point mutations of hemoglobin genes were found including HBA1: codon 14 TGG > TGA, HBA1: IVS-I-65 A > G, HBA1: codon 139 AAA > ATA, HBA2:-43 G > C, HBA2: c.*136 A >G, HBB:-99 G >A, HBB:-96 G > A, HBB:-62 G> A, HBB: c.56 C > T and HBB: 3'UTR +133 G > C. Four of the 15 thalassemia patients were HBA1: codon 14 TGG > TGA carriers. Conclusions Ten new point mutations of hemoglobin genes are detected, which enrich the hemoglobin gene mutation database. HBA1: codon 14 TGG>TGA may be a common alpha-thalassemia mutation in the population of Longyan, Fujian Province. Subsequent investigations with large sample size are urgently required to determine the carrying rate of HBA1: codon 14 TGG > TGA in thalassemia patients from this area.
引文
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[10]庞婉容,龙驹,叶学和,唐维骏,孙雷,劳可干,颜善活.广西北部湾地区人群地中海贫血基因突变分析.中国优生与遗传杂志,2016,24(1):39-42.
[11]黄烁丹,邹婕,庄宇嫦,熊蓉,张小燕,郑炎,邱美兰.广东梅州地区地中海贫血基因突变类型分析.新医学,2016,47(4):261-265.
[12]刘兴梅,苏莉,李贵芳,吴娴,王茹蕾,黄盛文.贵州地区重型β-地中海贫血基因突变类型分析.贵阳医学院学报,2016,41(4):407-409.
[2]Harteveld CL,Higgs DR.Alpha-thalassaemia.Orphanet J Rare Dis,2010,5:13.
[3]阎志杰,吴冠芸,王荣新,张俊武,刘敬忠,黄有文.非缺失型HbH病基因突变类型的研究.中华血液学杂志,1994,15(8):393-395.
[4]Cao A,Galanello R.Beta-thalassemia.Genet Med,2010,12(2):61-76.
[5]Harteveld CL,Yavarian M,Zorai A,Quakkelaar ED,van Delft P,Giordano PC.Molecular spectrum of alpha-thalassemia in the Iranian population of Hormozgan:three novel point mutation defects.Am J Hematol,2003,74(2):99-103.
[6]吴蓓颖,江岑,王也飞,顾燕英,林琳,蔡刚.血常规、血清铁及血红蛋白电泳联合检测在地中海贫血非高发地区的筛查意义.中华血液学杂志,2016,37(10):908-911.
[7]周常文,李厚钧,余伍忠,郝小军.新疆一例汉族家系β地中海贫血IVS-Ⅱ-654(C→T)的基因分析.八一农学院学报,1993,16(4):52-55.
[8]于洁,宪莹,姚秀云,肖剑文,刘海燕,陈仕平,张磊,张渝美,秦蓁子.重庆市学龄前儿童α地中海贫血的分子流行病学研究.中华血液学杂志,2014,35(5):419-423.
[9]张莉,黄伟忠,唐景云,黄国瑜,叶国永,喻长顺.联合应用MLPA和测序技术检测地中海贫血基因缺陷.实用预防医学,2014,21(7):779-781.
[10]庞婉容,龙驹,叶学和,唐维骏,孙雷,劳可干,颜善活.广西北部湾地区人群地中海贫血基因突变分析.中国优生与遗传杂志,2016,24(1):39-42.
[11]黄烁丹,邹婕,庄宇嫦,熊蓉,张小燕,郑炎,邱美兰.广东梅州地区地中海贫血基因突变类型分析.新医学,2016,47(4):261-265.
[12]刘兴梅,苏莉,李贵芳,吴娴,王茹蕾,黄盛文.贵州地区重型β-地中海贫血基因突变类型分析.贵阳医学院学报,2016,41(4):407-409.