Single nucleotide polymorphism genotyping of ALDH2 gene based on asymmetric PCR and fluorescent probe-mediated melting curves

作者全名："Zhang, Limei; Liu, Dan; Li, Baolin; Xie, Jingling; Liu, Jinbo; Zhang, Zhang"

作者地址："[Zhang, Limei; Li, Baolin; Xie, Jingling; Liu, Jinbo; Zhang, Zhang] Southwest Med Univ, Dept Lab Med, Affiliated Hosp, Luzhou 646000, Peoples R China; [Liu, Dan] Zigong Fourth People Hosp, Dept Clin Lab, Zigong 643099, Peoples R China; [Zhang, Zhang] Chongqing Med Univ, Dept Lab Med, Key Lab Lab Med Diagnost Educ, Chongqing 400010, Peoples R China"

通信作者："Liu, JB; Zhang, Z (通讯作者)，Southwest Med Univ, Dept Lab Med, Affiliated Hosp, Luzhou 646000, Peoples R China."

来源：ANALYTICAL BIOCHEMISTRY

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000793111800010

JCR分区：Q2

影响因子：2.9

年份：2022

卷号：642

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：ALDH2 polymorphism; Melting curve; Genotyping; Asymmetric PCR

摘要："Detection of single nucleotide polymorphisms (SNPs) is of great value in precision medicine. The polymorphism of the aldehyde dehydrogenase 2 (ALDH2) gene is caused by a G1510A transition, resulting in the substitution of glutamic acid by lysine at position 487. People of different ALDH2 genotypes show different susceptibility to cancer, metabolic diseases, etc. SNP analysis based on fluorescent probe-mediated melting curves is a relatively efficient and cost-effective method. Genomic DNA extracted from 100 whole blood samples was subjected to polymorphisms mutational analysis using asymmetric PCR and probe-mediated melting curves. Then a certain number of samples from each genotype were randomly selected for direct sequencing verification. The new assay can be performed in 2 h without post-PCR processing such as gel electrophoresis and validated by direct sequencing in a blind study with 100% concordance. Moreover, comparing the detection of polymorphisms of ALDH2 with the clinics, and an overall agreement of 100% (100/100) was demonstrated. Our study has shown a high level of concordance between DNA sequencing, which is suitable for the detection of clinical specimens. Based on the concept of probe-mediated melting curves, we further developed this platform as a universal strategy for the detection of polymorphisms related to folate metabolism."

基金机构：Affiliated Hospital of Southwest Medical University; National Natural Science Foundation of China [81802110]

基金资助正文：Biospecimens used in this study were provided by The Affiliated Hospital of Southwest Medical University. This work was supported by the National Natural Science Foundation of China (Grant No. 81802110) .