CAJ | 학술논문

目的探讨多重连接依赖性探针扩增(multiplex ligation-dependent probe amplification,MLPA)技术在α地中海贫血(简称地贫)产前基因诊断中的应用.方法对2例血液学检查(全血细胞计数和血红蛋白分析)为α地贫表型特征,α地贫基因常规方法检测为正常的孕妇的血液样本进行MLPA检测;对89对夫妇双方或一方为α地贫基因携带者的家系中的胎儿样本,采用常规跨越断裂点PCR(Gap polymerase chain reaction,Gap-PCR)技术和反向斑点杂交技术检测α珠蛋白基因的缺失及点突变;同时应用MLPA技术对上述89份样本进行α珠蛋白基因缺失检测.结果上述2例孕妇的血液样本,经MLPA检测未检测到α珠蛋白基因缺失,后经证实为缺铁性贫血;88份胎儿样本的MLPA检测结果和Gap-PCR检测结果完全一致;1份胎儿样本(绒毛组织)经Gap-PCR检测未能确诊,后结合MLPA检测确诊为-SEA/αα.结论 MLPA技术可应用于α地贫的产前基因诊断,作为Gap-PCR技术的有效补充,减少漏诊和误诊,提高产前诊断的准确率.
목적 탐토다중련접의뢰성탐침확증(multiplex ligation-dependent probe amplification,MLPA)기술재α지중해빈혈(간칭지빈)산전기인진단중적응용.방법 대2례혈액학검사(전혈세포계수화혈홍단백분석)위α지빈표형특정,α지빈기인상규방법검측위정상적잉부적혈액양본진행MLPA검측;대89대부부쌍방혹일방위α지빈기인휴대자적가계중적태인양본,채용상규과월단렬점PCR(Gap polymerase chain reaction,Gap-PCR)기술화반향반점잡교기술검측α주단백기인적결실급점돌변;동시응용MLPA기술대상술89빈양본진행α주단백기인결실검측.결과 상술2례잉부적혈액양본,경MLPA검측미검측도α주단백기인결실,후경증실위결철성빈혈;88빈태인양본적MLPA검측결과화Gap-PCR검측결과완전일치;1빈태인양본(융모조직)경Gap-PCR검측미능학진,후결합MLPA검측학진위-SEA/αα.결론 MLPA기술가응용우α지빈적산전기인진단,작위Gap-PCR기술적유효보충,감소루진화오진,제고산전진단적준학솔.
Objective To assess the application value of multiplex ligation-dependent probe amplification (MLPA) for the detection of gene deletion and prenatal diagnosis of a-thalassemia.Methods MLPA was applied for 2 cases with α-thalassemia phenotype by whole blood cell counting and hemoglobin component detection but were ruled out by regular molecular diagnosis.Potential gene deletions and point mutations of α-thalassemia gene were detected with regular Gap-polymerase chain reaction(Gap-PCR)and reverse dot blotting (RDB) in 89 cases where one or both partners were carriers of α-thalassemia mutations.Meanwhile, MLPA was used for detecting α-globin gene deletion among the 89 samples.Results For the 2 cases with α-thalassemia phenotype, no alpha globin gene deletion was detected by MLPA, but were subsequently confirmed as iron-deficiency anemia.The results of MLPA and Gap-PCR detection for the 88 cases were consistent, except for 1 fetal sample (chorionic villi) which could not be diagnosed by Gap-PCR and was confirmed to be--SEA/αα by MLPA.Conclusion MLPA can be applied to prenatal diagnosis of α-thalassemia as an effective supplement to Gap-PCR to reduce both misdiagnosis and missed diagnosis and improve the accuracy of prenatal diagnosis.