[1]郝文霞,张阳东,宋丽杰,等.核酸质谱技术对辐射相关基因单核苷酸多态性多重检测方法的构建及验证[J].现代检验医学杂志,2023,38(05):5-11.[doi:10.3969/j.issn.1671-7414.2023.05.002]
　HAO Wenxia,ZHANG Yangdong,SONG Lijie,et al.Establishment and Validation of A Multiple Detection Method for Single Nucleotide Polymorphisms of Genes Associated with Radiation Using Nucleic Acid Mass Spectrometry Technology[J].Journal of Modern Laboratory Medicine,2023,38(05):5-11.[doi:10.3969/j.issn.1671-7414.2023.05.002]
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核酸质谱技术对辐射相关基因单核苷酸多态性多重检测方法的构建及验证()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年05期
页码:
5-11
栏目:
论著
出版日期:
2023-09-15

文章信息/Info

Title:
Establishment and Validation of A Multiple Detection Method for Single Nucleotide Polymorphisms of Genes Associated with Radiation Using Nucleic Acid Mass Spectrometry Technology
文章编号:
1671-7414（2023）05-005-07
作者:
郝文霞1; 张阳东1; 宋丽杰1; 高 峥2; 牛冬梅2; 楚瑞雪3
（1. 火箭军特色医学中心，北京100088；2. 火箭军96606 部队医院，河南洛阳 471003；3. 解放军总医院京中医疗区礼士路门诊部，北京 100820）
Author(s):
HAO Wenxia1;  ZHANG Yangdong1;  SONG Lijie1;  GAO Zheng2;  NIU Dongmei2;  CHU Ruixue3
（1. the PLA Rocket Force Characteristic Medical Center, Beijing 100088, China; 2. Rocket Army 96606 Military Hospital, Henan Luoyang 471003，China; 3. Lishilu Clinic, Central Medical Branch of PLA General Hospital, Beijing 100820, China）
关键词:
电离辐射; 单核苷酸多态性; 核酸质谱技术
分类号:
Q503
DOI:
10.3969/j.issn.1671-7414.2023.05.002
文献标志码:
A
摘要:
目的 构建可同时检测多个辐射相关基因单核苷酸多态性（single nucleotide polymorphism, SNP）的高通量、高分辨率、快速的方法。方法 选取2020 年4 ～ 6 月火箭军特色医学中心75 例年满18 周岁的健康男性为研究对象，采集外周血并提取DNA。文献筛选辐射相关17 种基因28 个SNP，设计多重PCR 引物和单碱基延伸引物，通过PCR 扩增和延伸，应用DP-TOF 型飞行时间质谱检测系统对SNP 位点进行基因分型，并对其中7 份样本用Sanger 测序法进行验证。结果 构建的28 个SNP 均分型成功。75 份DNA 样本28 个SNP 的基质辅助激光解吸电离飞行时间质谱（martix-assistedlaser desorption/ionization time of flight mass spectrometry, MALDI-TOF-MS）检测结果显示基因分型检出率为100.0%。7 份DNA 样本28 个SNP 的基因型与Sanger 测序结果完全符合，其准确度为100.0%。结论 成功构建可同时检测17种基因28 个辐射相关SNP 的核酸质谱检测技术平台，该方法具有高通量、快速准确的优点。
Abstract:
Objective To establish a high-throughput, high-resolution and fast method that can simultaneously detect multiple single nucleotide polymorphisms (SNPs) of genes associated with radiation. Methods 75 healthy males over the age of 18 at the PLA Rocket Force Characteristic Medical Center from April 2020 to June 2020 were selected as the research subjects to collect peripheral blood and extract DNA. 17 genes with 28 SNPs were screened from literature. Multiple PCR primers and single base extension primers were designed. Genotypes of SNPs were tested with the DP-TOF time-of-flight mass spectrometry detection system after PCR amplification and extension, and 7 of them were verified by Sanger sequencing method. Results The 28 SNPs constructed were genotyped successfully. 28 SNPs in 75 DNA samples detected by(martix-assisted laser desorption/ionization time of flight mass spectrometrym, MALDI-TOF-MS) showed a genotyping detection rate of 100.0%. The genotyped of 28 SNPs from 7 samples were completely consistent with Sanger’s sequencing results, with an accuracy of 100.0%. Conclusion The nucleic acid mass-spectrometric detection technology platform which could simultaneously detect 28 SNPs of 17 genes associated with radiation was successfully constructed. It has the advantages of high-throughput, fast and accuracy.

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备注/Memo

备注/Memo:
基金项目：军队后勤面上项目（CEP21J009）；中心创新课题（ZXKC19019）。
作者简介：郝文霞（1989-），女，硕士研究生，医师，研究方向：基础医学研究，E-mail: hwx14551@163.com。
通讯作者：楚瑞雪（1970-），女，本科，副主任医师，研究方向：基础医学研究和健康管理，E-mail: churuixue666@163.com。
更新日期/Last Update: 2023-09-15