CAJ | 학술논문

目的了解近年来广东地区人群中α、β珠蛋白基因的基因型分布,并对β珠蛋白基因的单核苷酸多态性(SNP)进行研究,为更准确地进行地中海贫血的遗传诊断提供理论依据.方法利用单管多重PCR检测3种α缺失型地中海贫血,采用反向杂交(RDB)/PCR技术检测αCS、αQS、αWS3种α非缺失型及中国人常见的18种β珠蛋白的基因突变,对454份广东籍受检者标本进行地中海贫血基因分型,对其中的142份标本进行变性高效液相色谱(DHPLC)分析和β珠蛋白全基因测序,分析广东地区人群β珠蛋白基因SNP.结果在438例基因分析确诊的地中海贫血患者中,α、β、αβ复合型地中海贫血分别为246、164和28例.246例α地中海贫血患者中,检出αα/--SEA197例、αα/-α3.7 20例、αα/-α4.2 9例.164例β地中海贫血中,152例(92.7％)为杂合子,以CD41-42、IVS-Ⅱ-654、-28和CD17为主；8例(4.9％)为复合杂合子,4例(2.4％)为纯合子.DHPLC结合测序筛查β珠蛋白基因与RDB诊断结果一致,并发现9种SNP,其中IVS-Ⅰ-13 G＞A、IVS-Ⅱ-310 T＞C未见报道.在检测标本中,有4种SNP频率较高,73.2％的患者同时合并其中3种SNP (rs713040、rs10768683、rs1609812).结论在α地中海贫血中,主要以αα/-SEA基因型为主；β地中海贫血以CD41-42、IVS-Ⅱ-654、-28、CD17为主要突变类型；β地中海贫血复合杂合子、纯合子及αβ复合型地中海贫血的频率较高.DHPLC结合β珠蛋白全基因测序可以快速、有效地检出常见β地中海贫血突变型.rs713040、rs10768683、rs7480526、rs1609812在筛查人群中频率较高.
목적 료해근년래엄동지구인군중α、β주단백기인적기인형분포,병대β주단백기인적단핵감산다태성(SNP)진행연구,위경준학지진행지중해빈혈적유전진단제공이론의거.방법 이용단관다중PCR검측3충α결실형지중해빈혈,채용반향잡교(RDB)/PCR기술검측αCS、αQS、αWS3충α비결실형급중국인상견적18충β주단백적기인돌변,대454빈엄동적수검자표본진행지중해빈혈기인분형,대기중적142빈표본진행변성고효액상색보(DHPLC)분석화β주단백전기인측서,분석엄동지구인군β주단백기인SNP.결과 재438례기인분석학진적지중해빈혈환자중,α、β、αβ복합형지중해빈혈분별위246、164화28례.246례α지중해빈혈환자중,검출αα/--SEA197례、αα/-α3.7 20례、αα/-α4.2 9례.164례β지중해빈혈중,152례(92.7％)위잡합자,이CD41-42、IVS-Ⅱ-654、-28화CD17위주；8례(4.9％)위복합잡합자,4례(2.4％)위순합자.DHPLC결합측서사사β주단백기인여RDB진단결과일치,병발현9충SNP,기중IVS-Ⅰ-13 G＞A、IVS-Ⅱ-310 T＞C미견보도.재검측표본중,유4충SNP빈솔교고,73.2％적환자동시합병기중3충SNP (rs713040、rs10768683、rs1609812).결론 재α지중해빈혈중,주요이αα/-SEA기인형위주；β지중해빈혈이CD41-42、IVS-Ⅱ-654、-28、CD17위주요돌변류형；β지중해빈혈복합잡합자、순합자급αβ복합형지중해빈혈적빈솔교고.DHPLC결합β주단백전기인측서가이쾌속、유효지검출상견β지중해빈혈돌변형.rs713040、rs10768683、rs7480526、rs1609812재사사인군중빈솔교고.
Objective To understand the genotype of α and β-globin,as well as the polymorphism of β-globin gene in Cantonese in recent years,and to provide an effective genetic diagnosis for thalassemia (thal).Methods The single-tube complex PCR was used to detect 3 types of deletional α-thal,reverse dot blotting (RDB)/PCR to detect 3 kinds of undeletional α-thal-αCS,αQS,αWSand 18 kinds of β-thal mutations which were common in Chinese population.A total of 454 cases from Guangdong were undergone thal genotype genetic diagnosis.Among the 454 cases,142 cases were selected to perform the single nucleotide polymorphisms (SNPs) analysis of β-globin gene by denaturing high-performance liquid chromatography (DHPLC) combining the whole gene sequencing.Results Of the 454 cases,438 were diagnosed as thalassemia,including 246 of α-thal,164 of β-thal and 28 of αβ-thal.In 246 α-thal cases,deletions were the dominant mutations,including 197 cases of αα/--SEA,20 of αα-α3.7 and 9 of αα/-α4.2.In 164 β-thal cases,heterozygotes accounted for 92.7％ (152/164),the main genotypes were CD41-42,IVS-Ⅱ-654,-28 and CD17,and the dual heterozygotes and homozygotes accounted for 4.9％ (8/164) and 2.4％ (4/164),respectively.The result of β-globin gene screening by DHPLC combining with sequencing was consistent with that of RDB.Moreover,we also found 9 kinds of SNP,in which 2 were unreported,the IVS-Ⅰ-13 G＞A and IVS-Ⅱ-310 T＞C.In the tested samples,the frequency of 4 kinds SNP was high,among which 3 kinds SNPs-rs713040,rs10768683 and rs1609812 were carried together.Conclusion The dominant genotypes were αα/--SHA in α-thal cases,CD41-42,IVS-Ⅱ-654,-28 and CD 17 in β-thal.The frequency of β-thal heterozygotes,homozygotes and αβ-thal is high.DHPLC combining the whole β-globin gene sequencing can effectively detect the common β-thal mutation and even new mutations or SNPs.In Cantonese,the frequency of SNP rs713040,rs10768683,rs7480526 and rs1609812 of β-globin gene was high,and there may exist genetic linkage between rs713040,rs10768683 and rs1609812.