CAJ | 학술논문

目的探讨多重连接依赖式探针扩增(M LPA)技术在α地中海贫血(地贫)基因缺失检测及产前诊断中的应用.方法采用全血细胞计数和血红蛋白成分检测对受检者外周血标本进行表型分析,采用常规跨越裂点PCR(gap-PCR)技术及反向斑点杂交(RDB)法检测α地贫基因缺失及点突变,采用MLPA技术检测α珠蛋白基因缺失；产前诊断采用gap-PCR和MLPA法对胎儿标本进行α地贫基因缺失检测.结果 1256份(628对夫妇)受检者外周血标本中,75份检出α地贫表型特征.其中71份经常规方法检出α地贫基因突变且与表型相符；3份经常规方法未检出基因突变和1份具有血红蛋白H病表型且基因型为-α4.2/αα的标本,经MLPA法各检出1例α珠蛋白基因簇大片段缺失；筛出17对高风险夫妇.17份产前诊断标本中,2份受外源DNA污染的绒毛标本经MLPA法获得确诊.结论 MLPA是α地贫基因缺失常规gap-PCR检测方法的有效补充,可防止α地贫基因缺失的漏诊及产前诊断中的假阳性或假阴性误诊.
목적 탐토다중련접의뢰식탐침확증(M LPA)기술재α지중해빈혈(지빈)기인결실검측급산전진단중적응용.방법 채용전혈세포계수화혈홍단백성분검측대수검자외주혈표본진행표형분석,채용상규과월렬점PCR(gap-PCR)기술급반향반점잡교(RDB)법검측α지빈기인결실급점돌변,채용MLPA기술검측α주단백기인결실；산전진단채용gap-PCR화MLPA법대태인표본진행α지빈기인결실검측.결과 1256빈(628대부부)수검자외주혈표본중,75빈검출α지빈표형특정.기중71빈경상규방법검출α지빈기인돌변차여표형상부；3빈경상규방법미검출기인돌변화1빈구유혈홍단백H병표형차기인형위-α4.2/αα적표본,경MLPA법각검출1례α주단백기인족대편단결실；사출17대고풍험부부.17빈산전진단표본중,2빈수외원DNA오염적융모표본경MLPA법획득학진.결론 MLPA시α지빈기인결실상규gap-PCR검측방법적유효보충,가방지α지빈기인결실적루진급산전진단중적가양성혹가음성오진.
Objective To investigate the multiplex ligation-dependent probe amplification (MLPA) technology in the detection of gene deletion and prenatal diagnosis of α-thalassaemia.Methods Phenotypes were analyzed by whole blood cell counting and hemoglobin component detection of peripheral blood samples from the subjects.The gene deletions and point mutations of α-thalassaemia were detected with regular gap-PCR and reverse dot blot (RDB) method.At last,the MLPA method was applied for detection of α-globin gene deletion.All the prenatal diagnosis samples were detected with both gap-PCR and MLPA method.Results α-thalassaemia phenotype was found in 75 samples from 1256 (628 couples) peripheral blood samples for pre-pregnancy or prenatal thalassemia gene screening.Among them,71 samples carrying α-gene mutations and consistent with phenotypes were detected by routine methods.Inthe other 3 samples with no α-gene mutations detected and 1 sample with HbH phenotype but genotype of-α42/αα were analyzed by MLPA and found each one samples of whole α-globin gene cluster deletion,respectively.Seventeen high risk couples were screened.Among the 17 prenatal diagnosis samples,2 villus samples contaminated by exogenous DNA were confirmed by MLPA method.Conclusions MLPA is an effective complement for α-thalassaemia gene deletion detection.The molecular diagnosis strategy and process of gap-PCR combined with MLPA for α-thalassaemia gene deletion detection can prevent the missing of gene deletion,and false-positive or false-negative misdiagnosis of α-thalassaemia in prenatal diagnosis.