基因释放剂结合异源双链分析快速检测β地贫最常见的一种突变

Heterodupiex analysis and genereleaser for rapid detection of the most common B-thalassemia mutation.

β地贫是一种分子病理具有高度异质性的、我国南方最为常见的遗传性血液病.在已发现的23种β地贫突变中,CD41-42(-TCTT)移码突变基因频率最高(达41.6%).根据PCR原理和该突变的特点,作者设计了一对引物进行PCR特异扩增,电泳分析即可检出此种突变.并引进基因释放剂DNA模板快速制备法,从而使此技术更为简便和实用.作者运用此法检测了广东珠海地区114份β地贫样品,CD41-42突变捡出率为40.35%,并成功地应用于产前诊断中.根据实验结果,作者建议在分析未知β地贫突变时,首先采用本法先对CD41-42突变进行初筛,而将RDB和ASO技术作为补充.

Codon (CD) 41-42(-TCTT) mutation of 23 β-thalassemia mutations in China has been the most common, gene frequency of this allele is 41. 6 per cent. Based on the fact that above-mentioned, a set of primers were used for amplifing specially with PCR the 93 bp fragment of β-globin gene which encompass the deletion site, after genere-leaser was employed as rapid preparation of genom-ic DNA from blood and amniotic fluid cells. PCR products of CD 41-42 heterozygote were elec-trophoresed in a polyarcylamide gel, two heterodu-plexes were detected in addition to the expected band, giving the characteristic electrophoresis pattern. A simple, rapid approach was worked out to distinguish the homozygote and heterozygote of the mutation or normal individual and successfully applied to detection of 46 in 114 unknown β-thalass-mia samples and the prental diagnosis of a fetus at risk.