基础医学与临床2024，Vol.44Issue(6)：858-865,8.DOI:10.16352/j.issn.1001-6325.2024.06.0858
基于直接TaqMan-PCR的无创检测脂质代谢相关SNP基因分型技术
Direct TaqMan-PCR based technology for non-invasive genotyping of lipid metabolism associated SNPs
孙元宏 1谢吕 1范利华 1郑直1
作者信息
- 1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生物化学与分子生物学系,北京 100005
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摘要
Abstract
Objective To establish a non-invasive single nucleotide polymorphism genotyping technology based on direct TaqMan-PCR for high-throughput genotyping of site rs688 and rs964184 associated with lipid metabolism to meet the need for early screening of at-risk populations.Methods Extracted DNA was used to optimize the PCR annealing extension temperature and amplification procedure for the designed TaqMan probe;the oral swabs were placed into the sample treatment solution and then briefly centrifuged,and a small amount of the supernatant was taken for the direct qPCR amplification analysis;the freeze-thawing stability of probe and the effects of sample stor-age time on genotyping results were explored.Results The technology requires only simple treatment of oral swab for PCR analysis and can be stored at room temperature for 4 d.The optimized method was able to distinguish homo-zygous wild type,homozygous mutant type and heterozygote at rs688 and rs964184 loci.Conclusions A fast,con-venient,high-throughput,and low-cost SNP genotyping method has been established,providing an efficient and accurate new approach for large-scale screening of high-risk populations carrying susceptibility genes.关键词
直接PCR/核酸提取/SNP基因分型/高通量Key words
direct PCR/nucleic acids extraction/SNP genotyping/high-throughput分类
医药卫生引用本文复制引用
孙元宏,谢吕,范利华,郑直..基于直接TaqMan-PCR的无创检测脂质代谢相关SNP基因分型技术[J].基础医学与临床,2024,44(6):858-865,8.
