双重实时荧光PCR与DNA测序分析及PCR-RFLP方法检测白细胞介素-6基因启动子区多态性方法的比较研究

Comparative Studies on Detecting Method for Genotyping the Promoter Polymorphisms of IL-6 Gene by Duplex Real-time PCR,DNA Sequence Analysis and PCR-RFLP

目的比较双重实时荧光PCR与DNA测序分析以及PCR-RFLP三种检测方法对同一样本检测结果的一致性。方法分别采用三种方法对77例样本IL-6基因启动子区-572G/C,-597G/A 2个位点多态性进行检测。结果在77例样本中,采用双重实时荧光PCR方法与DNA测序分析相比仅有3例结果不一致。用限制性内切酶BsrBI酶切,有20例与实时荧光PCR方法结果不一致。当PCR产物用限制性内切酶Fok I酶切时,仅有5例被发现是GA型,另外两例未被检测出。结论双重实时荧光PCR方法与其它方法比较具有简单、准确、快速的特点,便于临床实验室大规模样本筛查。

Objective To compare the concordance for the same sample by duplex real-time PCR,DNA sequence analysis and PCR-RFLP. Methods The -572G/C and -597G/A promoter polymorphisms of the IL-6 gene in 77 samples by these three methods. Results Compared duplex real-time PCR method with DNA sequence analysis,three samples were different only in 77 samples. However,when the PCR product of 77 samples was digested by restriction enzyme BsrBI,20 of them were different from duplex real-time PCR method. The PCR product was digested by restriction enzyme Fok I,only five cases were found to be the GA genotype,as the other two cases were lost. Conclusion Compared with PCR-RFLP analysis and DNA sequencing analysis ,duplex real-time PCR is a simple ,accurate ,and fast method ,which is more suitable for large-sealed screen in clinical laboratory.