论文部分内容阅读
目的建立基于PCR-RDB技术的可同时检测中国人群G6PD最常见的4种基因突变类型的基因分型方法并对其作出评价。方法分别针对中国人群G6PD最常见的4种基因突变类型95(A→G)、1376(G→T)、1388(G→A)、1024(C→T)设计PCR扩增引物和基因分型探针,将探针固定到尼龙膜上制做成杂交膜条,建立PCR扩增体系和杂交体系。利用13例已知基因型的样本建立针对这4种基因突变类型的PCR-RDB检测方法。盲法对109例上述4种G6PD样本和正常样本进行PCR-RDB检测以及基因序列测定,并对两种方法所得结果进行对比。结果 PCR-RDB法准确地检测出了检测范围内所有的突变类型和正常样本,检测结果与测序结果完全一致。结论 PCR-RDB是一种经济、简便、高效、准确的G6PD基因分型方法 ,可用于G6PD的分型诊断和分型研究,并可做为一种常规方法推广到临床进行基因分型检测。
OBJECTIVE: To establish and evaluate a genotyping method based on PCR-RDB for simultaneous detection of the four most common G6PD gene mutations in Chinese population. Methods PCR amplification primers and genotypes were designed for the four most common mutations of G6PD in Chinese population (A → G), 1376 (G → T), 1388 (G → A), and 1024 (C → T) The probe is fixed on a nylon membrane to make a hybrid membrane membrane, and a PCR amplification system and a hybridization system are established. Using 13 samples of known genotypes, a PCR-RDB method was established to detect the mutation types of these four genes. Blind method PCR-RDB detection and gene sequencing of 109 cases of the above-mentioned 4 G6PD samples and normal samples were performed, and the results obtained by the two methods were compared. Results The PCR-RDB method accurately detected all the mutation types and normal samples in the detection range. The test results were in good agreement with the sequencing results. Conclusion PCR-RDB is an economical, simple, efficient and accurate method for genotyping G6PD. It can be used in the genotyping and typing of G6PD, and can be used as a routine method for clinical genotyping.