【摘要】 目的基因诊断结果在α-地贫的产前诊断中具有决定意义。采取两种不同技术原理的方法验证产前诊断结果,是保证产前诊断结果可靠性的基本要求。本文探讨检测常见非缺失型α-地贫的RDB技术作为单管多重PCR技术产前诊断α-地贫验证方法的可行性。方法采用双盲法,对73例α-地贫产前诊断的标本分别采用针对中国人3种常见缺失型α-地贫突变的单管多重gap-PCR技术和针对常见3种非缺失α-地贫点突变的RDB法进行分析,根据单管多重PCR技术扩增产物电泳图谱和RDB膜条上显色的斑点分别判断某一个标本缺失型和非缺失α-地贫的突变类型,且与引产或出生后胎儿脐带血中HbBart′s定量结果进行比对。结果单管多重PCR和RDB技术均能分别准确检出常见缺失型和非缺失型α-地贫各种突变类型。如将两者结合分析,还能提供相互佐证的重要信息:对于--SEA、-α3.7和-α4.2自身或相互组合的各种基因型标本,其RDB膜条的杂交斑点均不显色;而--SEA与αCSα、αQSα和αWSα组合的基因型标本该突变的正常对照点不显色。α-地贫产前诊断结果与胎儿脐带血电泳结果完全吻合。结论单管多重PCR结合RDB技术同时应用于α-地贫的基因诊断,可保证产前诊断的可靠性,值得在临床遗传诊断实验室中推广应用。更多还原

【Abstract】 Objective To study the feasibility of reverse dot blot (RDB) technology, used for the detection of common non-deletion type α-thalassemia(α-Thal), in the validation of single-tube multiplex PCR (STMP) for the prenatal diagnosis of α-Thal.Methods 73 specimens of prenatal diagnosis for α-Thal were detected simultaneously by STMP for three common deletion type of α-Thal (--SEA ,-α4.2 and -α3.7) and by RDB for three common non-deletion type of α-Thal (αCSα,αQSα and αWSα) by double blind trial. The genotypes of α-globin gene were acquired by the agarose gel electrophoresis profiles of STMP products for determining deletion type α-Thal and the color of spots on the RDB membrane strip for detecting non-deletion typeα-Thal. The results were confirmed further by quantificational levels of Hb Bart′s in the umbilical cord blood of fetus, delivered by induced abortion or normal delivery.Results STMP and RDB could accurately genotyped deletion and non-deletion type α-Thal mutations respectively. Combination of the results of both STMP and RDB could provide important information of corroborative evidence for each other: for combination of the above mentioned deletion type α-Thal themselves or each other, the hybridization spots on RDB membrane strip were with no color, but for --SEA combined with αCSα and αQSαcombined with αWSα, were colored only in the dot of non-deletion mutation probes. Both the results of prenatal diagnosis and the levels of Hb Bart′s in the umbilical cord blood were perfectly matched.Conclusion Simultaneously application of STMP PCR and RDB for the prenatal diagnosis of α-Thal could guarantee the reliability of result and could be worthy of deploying with confidence popularly in the clinical genetic laboratory.更多还原