国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2011年
2期
57-61,101
,共6页
石西南%姚莉琴%赵钟鸣%邹团标%史磊%于亮%褚嘉祐%杨昭庆
石西南%姚莉琴%趙鐘鳴%鄒糰標%史磊%于亮%褚嘉祐%楊昭慶
석서남%요리금%조종명%추단표%사뢰%우량%저가우%양소경
β-地中海贫血%β珠蛋白基因(HBB)%基因变异%序列分析
β-地中海貧血%β珠蛋白基因(HBB)%基因變異%序列分析
β-지중해빈혈%β주단백기인(HBB)%기인변이%서렬분석
Beta-thalassemia%Mutation%Hemoglobin beta gene%Sequence analysis
目的 探讨云南傣族人群中β-地中海贫血β珠蛋白基因(hemoglobin beta gene,HBB)变异分布特点.并建立检测相应常见变异位点的快速筛查方法.方法 获取209例云南傣族儿童β-地中海贫血阳性样本,用DNA序列测定的方法对包含有β珠蛋白基因(HBB)在内的1755bp DNA片段进行基因变异分析.建立检测3个常见变异位点的碱基突变特异性扩增系统(amplification refractory mutation system,ARMS),并对引物及扩增条件进行优化.结果 209例样本中共发现9个变异位点,按频率由高到低分别是CD2 T＞C(50.72%)、CD26 G＞A(35.41%)、CD17 A＞T(12.92%)、ⅣS-Ⅱ-17 C＞G(12.44%)、ⅣS-Ⅱ-16 G＞C(11.96%)、ⅣS-Ⅰ-30 A＞G(9.57%)、CD6G＞A(9.09%)、CD41-42(-TCTT)(7.18%)、CD5 T＞A(1.92%),有153人被检测出基因变异位点,占总人数的73.21%.引入错配碱基的ARMS引物,在65℃的退火温度能对CD2、CD26和CD17 3个常见变异位点进行特异筛查.结论 云南傣族β-地中海贫血人群中HBB基因的变异位点有其自身的特点,与其他地区有所不同.ARMS检测云南傣族β-地中海贫血基因变异是一种准确有效并简便经济的方法.
目的 探討雲南傣族人群中β-地中海貧血β珠蛋白基因(hemoglobin beta gene,HBB)變異分佈特點.併建立檢測相應常見變異位點的快速篩查方法.方法 穫取209例雲南傣族兒童β-地中海貧血暘性樣本,用DNA序列測定的方法對包含有β珠蛋白基因(HBB)在內的1755bp DNA片段進行基因變異分析.建立檢測3箇常見變異位點的堿基突變特異性擴增繫統(amplification refractory mutation system,ARMS),併對引物及擴增條件進行優化.結果 209例樣本中共髮現9箇變異位點,按頻率由高到低分彆是CD2 T＞C(50.72%)、CD26 G＞A(35.41%)、CD17 A＞T(12.92%)、ⅣS-Ⅱ-17 C＞G(12.44%)、ⅣS-Ⅱ-16 G＞C(11.96%)、ⅣS-Ⅰ-30 A＞G(9.57%)、CD6G＞A(9.09%)、CD41-42(-TCTT)(7.18%)、CD5 T＞A(1.92%),有153人被檢測齣基因變異位點,佔總人數的73.21%.引入錯配堿基的ARMS引物,在65℃的退火溫度能對CD2、CD26和CD17 3箇常見變異位點進行特異篩查.結論 雲南傣族β-地中海貧血人群中HBB基因的變異位點有其自身的特點,與其他地區有所不同.ARMS檢測雲南傣族β-地中海貧血基因變異是一種準確有效併簡便經濟的方法.
목적 탐토운남태족인군중β-지중해빈혈β주단백기인(hemoglobin beta gene,HBB)변이분포특점.병건립검측상응상견변이위점적쾌속사사방법.방법 획취209례운남태족인동β-지중해빈혈양성양본,용DNA서렬측정적방법대포함유β주단백기인(HBB)재내적1755bp DNA편단진행기인변이분석.건립검측3개상견변이위점적감기돌변특이성확증계통(amplification refractory mutation system,ARMS),병대인물급확증조건진행우화.결과 209례양본중공발현9개변이위점,안빈솔유고도저분별시CD2 T＞C(50.72%)、CD26 G＞A(35.41%)、CD17 A＞T(12.92%)、ⅣS-Ⅱ-17 C＞G(12.44%)、ⅣS-Ⅱ-16 G＞C(11.96%)、ⅣS-Ⅰ-30 A＞G(9.57%)、CD6G＞A(9.09%)、CD41-42(-TCTT)(7.18%)、CD5 T＞A(1.92%),유153인피검측출기인변이위점,점총인수적73.21%.인입착배감기적ARMS인물,재65℃적퇴화온도능대CD2、CD26화CD17 3개상견변이위점진행특이사사.결론 운남태족β-지중해빈혈인군중HBB기인적변이위점유기자신적특점,여기타지구유소불동.ARMS검측운남태족β-지중해빈혈기인변이시일충준학유효병간편경제적방법.
Objective To characterize the distribution of variation in hemoglobin beta gene ( hemoglobin beta gene, HBB )of β-thalassemia in Dai minority group in Yunnan province, and to establish a rapid and economical method for screening β-thalassemia mutation via optimizing ARMS technique.Methods DNA Sequencing was performed to detect variation in 1755 bp DNA fragment containing whole length of hemoglob beta gene ( 1606 bp) from 209 cases of Dai children with β-thalassemia. The primer design and PCR conditions of ARMS (amplification refractory mutation system ) have been optimized for detecting three frequent variations. Results Among 209 samples, there are 9 variations were detected:CD2 T＞ C (50.72%),CD26 G ＞A (35.41%),CD17 A ＞T(12.92%),IVS-Ⅱ-17 C ＞ G (12.44%), IVS-Ⅱ-16 G＞C(11.96%), ⅣS-Ⅰ-30 A＞G(9.57%) , CD6 G ＞A(9.09%) ,CD41-42 (-TCTT) (7. 18%) and CD5 T＞A(1.92%). Three mutations (CD21, CD35 and CD43) were first reported in China. There are 167, out of 209 participants (79.90%), carrying HBB gene variation. Three common mutations (CD2, CD26 and CD17 ) can be efficiently detected by combining condition of both ARMS mismatch primers and 65 ℃ of PCR annealling temperature. Conclusion The distribution of HBB gene variation in the Dai minority group in Yunnan province is different from those of other groups in China. ARMS is an effective, convenient and economical technique for rapid detection of gene mutations of β-thalassemia.
