单细胞巢式聚合酶链反应诊断β地中海贫血

Diagnosis ofβ-thalassemia by single cell nested polymerase chain reaction

目的探讨在单淋巴细胞水平准确诊断β地中海贫血(β地贫)的方法,为开展β地贫的植入前诊断(PGD)打下基础。方法获取正常人及β地贫基因携带者的单个淋巴细胞,采用多重巢式聚合酶链反应(PCR)反向膜杂交(RDB)技术进行β地贫基因检测。结果单个淋巴细胞的PCR扩增效率为90.4%,等位基因脱扣(ADO)发生率为8.5%。结论用单细胞多重巢式PCR+RDB诊断β地贫是稳定可靠的,可望用于β地贫的临床植入前诊断。

Objective To develop a reliable and accurate method to diagnose β-thalassemia based on a single cell. Methods Improved multiplex nested polymerase chain reaction （PCR） and reverse dot blot （RDB） were used to detect the β-thalassemia gene from a single lymphocyte derived from the normal person and the β-thalassemia carrier. Result The PCR amplification efficiency of a single lymphocyte was 90.4% ,and the allele dropout rate was 8.5%. Conclusion The diagnosis of β-thalassemia based on a single cell by multiplex nested PCR and RDB is stable and reliable. It should be meaningful in preimplantation genetic diagnosis of β-thalassemia.