| 乙醇脱氢酶1B-rs1229984和乙醛脱氢酶2基因-rs671多态性实时荧光定量PCR检测方法的建立 |
| 投稿时间：2017-01-18 修订日期：2017-02-20 点此下载全文 |
| 引用本文：梁国威,邵冬华,何美琳,刘洁.乙醇脱氢酶1B-rs1229984和乙醛脱氢酶2基因-rs671多态性实时荧光定量PCR检测方法的建立[J].医学研究杂志,2017,46(11):139-144 |
| DOI：
10.11969/j.issn.1673-548X.2017.11.034 |
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| 中文摘要:目的 建立乙醇脱氢酶1B （ADH1B）基因rs1229984（G/A）和乙醛脱氢酶2基因rs671（G/A）多态性的实时荧光定量PCR分型方法。方法 分别采用双TaqMan-MGB探针法建立ADH1B-rs1229984多态性，和ARMS-TaqMan探针法建立ALDH2-rs671多态性的实时荧光定量PCR分型方法。根据荧光定量PCR的循环阈值（Ct值）的差值ΔCt值判定等位基因型，并采用所建方法对345例中国汉族人进行了分型检测。结果 所建方法对345例研究对象分型检测显示，ADH1B-rs1229984多态性的GG （ADH1B*1/*1）、GA （ADH1B*1/*2）和AA （ADH1B*2/*2）型的ΔCt值（ΔCt=G型TaqMan-MGB探针Ct值-A型TaqMan-MGB探针Ct值）分别为-13.72±1.33（40例）、0.50±0.17（150例）和6.62±0.54（155例）；ALDH2-rs671的GG （ALDH2*1/*1）、GA （ALDH2*1/*2）和AA （ALDH2*2/*2）型的ΔCt值（ΔCt=G引物反应体系Ct值-A引物反应体系Ct值）分别为-15.30±0.84（239例）、0.17±0.45（96例）和15.86±0.74（10例）。345例研究对象中，ADH1B*1/*2和ALDH2*1/*1（31.3%）、ADH1B*2/*2和ALDH2*1/*1（28.7%）是2个等位基因的主要组合型（60%），未见ADH1B*1/*1和ALDH2*2/*2组合型。结论 所建分型检测方法皆具有闭管、一步检测、准确和高通量等特点。 |
| 中文关键词:乙醇脱氢酶1B 乙醛脱氢酶2 基因多态性 荧光定量PCR |
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| Genotyping Methods for ADH1B-rs1229984 and ALDH2-rs671 Polymorphisms in a Real-time PCR |
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| Abstract:Objective To develop novel methods for genotyping alcohol dehydrogenase 1B gene (ADH1B)-rs1229984 (G/A) and aldehyde dehydrogenase 2 (ALDH2)-rs671 (G/A) polymorphisms.Methods The dual TaqMan-MGB probes for ADH1B-rs1229984 and amplification refractory mutation system combined a TaqMan probe (ARMS-TaqMan) for ALDH2-rs671 polymorphisms were used in developing the methods in a real-time PCR, respectively. The genotyping results were obtained by monitoring the difference of cycle threshold values (ΔCt) in the assays. Moreover, 345 Han subjects were genotyped for evaluating the established assays, respectively.Results In 345 subjects, The ΔCt (ΔCt=G allele TaqMan-MGB probe Ct values minus A allele TaqMan-MGB probe Ct values) for GG (ADH1B*1/*1), GA (ADH1B*1/*2) and AA (ADH1B*2/*2) of ADH1B-rs1229984 polymorphisms were -13.72±1.33 (40 cases),0.50±0.17 (150 cases) and 6.62±0.54 (155 cases), and the ΔCt (ΔCt=G allele-special primer Ct values minus A allele-special primer Ct values) for GG (ALDH2*1/*1), GA (ALDH2*1/*2) and AA (ALDH2*2/*2) of ALDH2-rs671 polymorphisms were -15.30±0.84 (239 cases),0.17±0.45 (96 cases) and 15.86±0.74 (10 cases), respectively. The main allelic combination types were ADH1B*1/*2 vs ALDH2*1/*1 (31.3%) and ADH1B*2/*2 vs ALDH2*1/*1 (28.7%), and no ADH1B*1/*1 vs ALDH2*2/*2 types were observed in 345 Han subjects.Conclusion Both of the novel assays had the features of accuracy, one step and high-throughput for the genotyping. |
| keywords:Alcohol dehydrogenase 1B Aldehyde dehydrogenase 2 Polymorphisms Real-time PCR |
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