中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2012年
1期
32-36
,共5页
孙顺昌%周指明%宋慧文%彭运生%贺敬波%莫宝妹
孫順昌%週指明%宋慧文%彭運生%賀敬波%莫寶妹
손순창%주지명%송혜문%팽운생%하경파%막보매
β地中海贫血%珠蛋白类%多态现象,遗传%突变
β地中海貧血%珠蛋白類%多態現象,遺傳%突變
β지중해빈혈%주단백류%다태현상,유전%돌변
beta-Thalassemia%Globins%Polymorphism,genetic%Mutation
目的 探讨轻型β-珠蛋白生成障碍性贫血患者β-珠蛋白基因(AC)n(AT)xTy多态性与其基因突变的相关性.方法 选取2009年2月至2010年7月深圳市宝安区人民医院89例已知基因突变类型的轻型β-珠蛋白生成障碍性贫血患者和110名中国汉族人群健康对照者.抽取所有个体外周静脉血,抽提基因组DNA,通过聚合酶链反应扩增β-珠蛋白基因BP1蛋白结合区序列,经DNA测序确定(AC)n(AT)xTy序列的多态性,分析(AC)n(AT)xTy多态性与其基因突变的关系.轻型β-珠蛋白生成障碍性贫血患者和健康对照者间(AC)n(AT)xTy多态性单倍型频率的比较,以及同一单倍型患者的不同突变类型发生率间的比较采用x2检验.结果 在轻型β-珠蛋白生成障碍性贫血患者的β-珠蛋白基因BP1蛋白结合区存在9种(AC)n(AT)xTy多态性序列,分别是(AC)2(AT)7T7、(AC)2 (AT)8T5、(AC)3 (AT)7T5、(AC)2 (AT)9T5、(AC)2 (AT)8T9、(AC)3 (AT)8T5、(AC)2(AT)10T3、(AC)2(AT)7T5和(AC)2(AT) 11T3,其中(AC)2 (AT)7T7和(AC)2(AT)8T5是常见单倍型.轻型β-珠蛋白生成障碍性贫血患者的(AC)2(AT)7T7、(AC)3 (AT)7T5和(AC)2(AT)8T9单倍型频率分别为38.8％( 69/178)、11.8％( 21/178)、9.0％( 16/178),显著高于健康对照组的24.1％(53/220)、5.4％ (12/220)、3.2％ (7/220),差异有统计学意义(x2=9.966、4.371、6.093,P＜0.05)；(AC)2 (AT)9T5单倍型频率为10.1％(18/178),显著低于健康对照组的33.2％ (73/220),差异有统计学意义(x2=29.691,P＜0.01)；而(AC)2 (AT)8T5单倍型频率在患者组和健康对照组分别为25.3％ (45/178)和29.1％ (64/220),差异无统计学意义(x2 =0.718,P＞0.05).在(AC)2 (AT)7T7单倍型患者中,codon41/42(-TTCT)和IVS-II-654(C→T)的突变率分别为59％( 10/17)和29％(5/17),差异无统计学意义(x2=2.982,P＞0.05)；在(AC)2( AT)8T5单倍型患者中,codon41/42(-TTCT)和IVS-Ⅱ-654(C→T)的突变率分别为29％(4/14)和57％ (8/14),差异无统计学意义(x2=2.333,P＞0.05).结论 β-珠蛋白基因BP1蛋白结合区的(AC)2 (AT) 7T7、(AC)3 (AT)7T5和(AC)2(AT)8T9单倍型与轻型β-珠蛋白生成障碍性贫血存在连锁不平衡.在轻型β-珠蛋白生成障碍性贫血患者中,携带( AC)2( AT)7T7单倍型和(AC)2(AT) 8T5单倍型患者的主要致病突变分别为codon41/42 (-TTCT)和IVS-II-654(C→T).
目的 探討輕型β-珠蛋白生成障礙性貧血患者β-珠蛋白基因(AC)n(AT)xTy多態性與其基因突變的相關性.方法 選取2009年2月至2010年7月深圳市寶安區人民醫院89例已知基因突變類型的輕型β-珠蛋白生成障礙性貧血患者和110名中國漢族人群健康對照者.抽取所有箇體外週靜脈血,抽提基因組DNA,通過聚閤酶鏈反應擴增β-珠蛋白基因BP1蛋白結閤區序列,經DNA測序確定(AC)n(AT)xTy序列的多態性,分析(AC)n(AT)xTy多態性與其基因突變的關繫.輕型β-珠蛋白生成障礙性貧血患者和健康對照者間(AC)n(AT)xTy多態性單倍型頻率的比較,以及同一單倍型患者的不同突變類型髮生率間的比較採用x2檢驗.結果 在輕型β-珠蛋白生成障礙性貧血患者的β-珠蛋白基因BP1蛋白結閤區存在9種(AC)n(AT)xTy多態性序列,分彆是(AC)2(AT)7T7、(AC)2 (AT)8T5、(AC)3 (AT)7T5、(AC)2 (AT)9T5、(AC)2 (AT)8T9、(AC)3 (AT)8T5、(AC)2(AT)10T3、(AC)2(AT)7T5和(AC)2(AT) 11T3,其中(AC)2 (AT)7T7和(AC)2(AT)8T5是常見單倍型.輕型β-珠蛋白生成障礙性貧血患者的(AC)2(AT)7T7、(AC)3 (AT)7T5和(AC)2(AT)8T9單倍型頻率分彆為38.8％( 69/178)、11.8％( 21/178)、9.0％( 16/178),顯著高于健康對照組的24.1％(53/220)、5.4％ (12/220)、3.2％ (7/220),差異有統計學意義(x2=9.966、4.371、6.093,P＜0.05)；(AC)2 (AT)9T5單倍型頻率為10.1％(18/178),顯著低于健康對照組的33.2％ (73/220),差異有統計學意義(x2=29.691,P＜0.01)；而(AC)2 (AT)8T5單倍型頻率在患者組和健康對照組分彆為25.3％ (45/178)和29.1％ (64/220),差異無統計學意義(x2 =0.718,P＞0.05).在(AC)2 (AT)7T7單倍型患者中,codon41/42(-TTCT)和IVS-II-654(C→T)的突變率分彆為59％( 10/17)和29％(5/17),差異無統計學意義(x2=2.982,P＞0.05)；在(AC)2( AT)8T5單倍型患者中,codon41/42(-TTCT)和IVS-Ⅱ-654(C→T)的突變率分彆為29％(4/14)和57％ (8/14),差異無統計學意義(x2=2.333,P＞0.05).結論 β-珠蛋白基因BP1蛋白結閤區的(AC)2 (AT) 7T7、(AC)3 (AT)7T5和(AC)2(AT)8T9單倍型與輕型β-珠蛋白生成障礙性貧血存在連鎖不平衡.在輕型β-珠蛋白生成障礙性貧血患者中,攜帶( AC)2( AT)7T7單倍型和(AC)2(AT) 8T5單倍型患者的主要緻病突變分彆為codon41/42 (-TTCT)和IVS-II-654(C→T).
목적 탐토경형β-주단백생성장애성빈혈환자β-주단백기인(AC)n(AT)xTy다태성여기기인돌변적상관성.방법 선취2009년2월지2010년7월심수시보안구인민의원89례이지기인돌변류형적경형β-주단백생성장애성빈혈환자화110명중국한족인군건강대조자.추취소유개체외주정맥혈,추제기인조DNA,통과취합매련반응확증β-주단백기인BP1단백결합구서렬,경DNA측서학정(AC)n(AT)xTy서렬적다태성,분석(AC)n(AT)xTy다태성여기기인돌변적관계.경형β-주단백생성장애성빈혈환자화건강대조자간(AC)n(AT)xTy다태성단배형빈솔적비교,이급동일단배형환자적불동돌변류형발생솔간적비교채용x2검험.결과 재경형β-주단백생성장애성빈혈환자적β-주단백기인BP1단백결합구존재9충(AC)n(AT)xTy다태성서렬,분별시(AC)2(AT)7T7、(AC)2 (AT)8T5、(AC)3 (AT)7T5、(AC)2 (AT)9T5、(AC)2 (AT)8T9、(AC)3 (AT)8T5、(AC)2(AT)10T3、(AC)2(AT)7T5화(AC)2(AT) 11T3,기중(AC)2 (AT)7T7화(AC)2(AT)8T5시상견단배형.경형β-주단백생성장애성빈혈환자적(AC)2(AT)7T7、(AC)3 (AT)7T5화(AC)2(AT)8T9단배형빈솔분별위38.8％( 69/178)、11.8％( 21/178)、9.0％( 16/178),현저고우건강대조조적24.1％(53/220)、5.4％ (12/220)、3.2％ (7/220),차이유통계학의의(x2=9.966、4.371、6.093,P＜0.05)；(AC)2 (AT)9T5단배형빈솔위10.1％(18/178),현저저우건강대조조적33.2％ (73/220),차이유통계학의의(x2=29.691,P＜0.01)；이(AC)2 (AT)8T5단배형빈솔재환자조화건강대조조분별위25.3％ (45/178)화29.1％ (64/220),차이무통계학의의(x2 =0.718,P＞0.05).재(AC)2 (AT)7T7단배형환자중,codon41/42(-TTCT)화IVS-II-654(C→T)적돌변솔분별위59％( 10/17)화29％(5/17),차이무통계학의의(x2=2.982,P＞0.05)；재(AC)2( AT)8T5단배형환자중,codon41/42(-TTCT)화IVS-Ⅱ-654(C→T)적돌변솔분별위29％(4/14)화57％ (8/14),차이무통계학의의(x2=2.333,P＞0.05).결론 β-주단백기인BP1단백결합구적(AC)2 (AT) 7T7、(AC)3 (AT)7T5화(AC)2(AT)8T9단배형여경형β-주단백생성장애성빈혈존재련쇄불평형.재경형β-주단백생성장애성빈혈환자중,휴대( AC)2( AT)7T7단배형화(AC)2(AT) 8T5단배형환자적주요치병돌변분별위codon41/42 (-TTCT)화IVS-II-654(C→T).
Objective To explore linkage relationship between polymorphisms of (AC)n (AT)xTy and mutations in the β-globin gene in patients with mild β-thalassemia.Methods The subjects were 89 mild β-thalassemia patients with known mutations and 110 healthy subjects from People's Hospital of Baoan District of Shenzhen from February 2009 to July 2010.Genomic DNA was extracted from peripheral leukocytes.Sequence of the BP1 binding site upstream of the β-globin gene was amplified by polymerase chain reaction,polymorphisms of (AC)n (AT)xTy were determined by DNA sequencing.Allelic frequencies of (AC)n (AT)xTy between mild β-thalassemia patients and healthy subjects were compared using x2 test.Mutation rates between two groups were also compared using x2 test for subjects carrying same haplotype. Linkage relationship was conducted according to allelic frequencies and mutations. Results Analysis of the (AC)n(AT) xTy polymorphisms of the BP1 binding site upstream of the β-globin gene showed 9 different genotypes: (AC)2( AT)7T7,( AC)2( AT)8T5,( AC)3( AT)7T5,( AC)2( AT)9T5,( AC)2(AT)8T9,(AC)3(AT)8T5,(AC)2(AT)10T3,(AC)2(AT)7T5 and (AC)2(AT)11T3.The (AC)2(AT)7T7 and (AC)2 (AT)8T5 genotypes were common for patients with mild β-thalassemia.Allele frequencies of (AC)2(AT)7T7,(AC)3 ( AT)7T5 and ( AC)2( AT)8T9 were 38.8％ (69/178),11.8％(21/178),9.0％ ( 16/178 ) for mild β-thalassemia patients,and 24.1％ ( 53/220),5.4％ ( 12/220),3.2％ (7/220) for healthy subjects, respectively, there were significant differences between mild β-thalassemia patients and healthy subjects (x2 =9.966,4.371,6.093,P ＜ 0.05 ).Allele frequency of (AC)2(AT)9T5 was 10.1％ (18/178) and 33.2％ (73/220) for mild β-thalassemia patients and healthy subjects,frequency of (AC)2 (AT)9T5 was significandy lower in mild β-thalassemia patients than in healthy subjects (x2 =29.691,P ＜0.01 ).Allele frequency of (AC)2(AT)8T5 was 25.3％ (45/178) and 29.1％(64/220) for mild β-thalassemia patients and healthy subjects,there wasn't significant difference between patients and healthy subjects (x2 =0.718,P ＞0.05).The mutation rates of codon41/42(-TTCT) and IVSⅡ-654(C→T) were 59％ (10/17) and 29％ (5/17) for mild β-thalassemia patients carrying (AC)2(AT)7T7 allele,and 29％ (4/14) and 57％ (8/14) for patients carrying ( AC)2 (AT)8T5 allele.There were not significant differences between codon41/42(-TTCT) mutation rate and IVS-Ⅱ-654(C→T) mutation rate (x2 =2.982,2.333,P ＞ 0.05 ) for mild β-thalassemia patients carrying ( AC)2 ( AT)7T7 and ( AC)2(AT)8T5 allele.Conclusions Allele of (AC)2(AT)7T7,(AC)3(AT)7T5 and (AC)2(AT)8T9 are in linkage disequilibrium with β-thalassemia.Most mild β-thalassemia patients carrying (AC)2 (AT)7T7 allele are caused by codon41/42 (-TTCT) mutation in the β-globin gene,and IVS-Ⅱ-654 (C→T) is a major mutation for patients carrying (AC)2(AT)8T5 allele.
