直接测序法与克隆测序法在单核苷酸多态性检测中的比较

A Comparison of Direct Sequencing with Clone Sequencing in the Detection of Single Nucleotide Polymorphism

目的：了解PCR产物直接测序法与克隆测序法在单核苷酸多态性（SNP）检测上的差别。方法：对2型糖尿病患者的载脂蛋白J外显子2基因及旁侧进行聚合酶链反应（PCR）扩增，分别对其产物进行直接测序和克隆测序。结果：在样本的检测中，PCR产物直接测序法所测序列与43～60bp以后克隆测序相同；检出一个突变位点并与GenBank（DQ012938）发表序列一致。结论：PCR产物直接测序法和克隆测序法都是检测SNP的有效方法，但前者不仅特异性强，敏感度高，而且比克隆测序方法更为快速简便，节省材料。

Objective： To compare the sensitivity detection of single nucleotide polymorphism（SNP） with polymerase chain reaction（PCR） in 8 patients and accuracy of the 2 sequencing techniques for the Methods： The exon 2 of Apo J gene was screened with type 2 diabetes mellitus （DM）. Samples were detected by direct sequencing and clone sequencing, and the results were compared. Results： One mutation was detected in exon 2 by two methods. Furthermore, it＇ s same to the Homo sapiens apolipoprotein J （CLU） gene sequence in GenBank （DQ012938）. Conclusion： The results show that the PCR direct sequencing is faster, easier, cheaper, and more practical than clone sequencing when used in SNP.