[1]吕晓玥,刘玉泉,惠文利.基于PCR-金磁纳米微粒层析技术检测乙醛脱氢酶2基因多态性方法的建立及应用[J].现代检验医学杂志,2017,32(04):21-24.[doi:10.3969/j.issn.1671-7414.2017.04.006]
　L(¨overU)Xiao-yue,LIU Yu-quan,HUI Wen-li.Establishment and Application of the Method of Polymorphism of Aldehyde Dehydrogenase 2 Gene Based on PCR-gold Magnetic Nanoparticle Chromatography[J].Journal of Modern Laboratory Medicine,2017,32(04):21-24.[doi:10.3969/j.issn.1671-7414.2017.04.006]
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基于PCR-金磁纳米微粒层析技术检测乙醛脱氢酶2基因多态性方法的建立及应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第32卷
期数:
2017年04期
页码:
21-24
栏目:
论著
出版日期:
2017-07-25

文章信息/Info

Title:
Establishment and Application of the Method of Polymorphism of Aldehyde Dehydrogenase 2 Gene Based on PCR-gold Magnetic Nanoparticle Chromatography
文章编号:
1671-7414(2017)04-021-04
作者:
吕晓玥1; 刘玉泉1; 惠文利2
1.咸阳市中心医院检验科,陕西咸阳 712000; 2.西北大学生命科学学院,西安 710069
Author(s):
L(¨overU)Xiao-yue1; LIU Yu-quan1; HUI Wen-li2
1.Department of Clinical Laboratory,Xianyang Central Hospital,Shaanxi Xianyang 712000, China; 2.Department of Biological Sciences,Northwest University,Xi'an710069,China
关键词:
乙醛脱氢酶2; 口腔拭子; 金磁微粒层析检测技术
分类号:
Q503
DOI:
10.3969/j.issn.1671-7414.2017.04.006
文献标志码:
A
摘要:
目的 基于“PCR-金磁纳米微粒层析法”建立用于口腔拭子样本类型的乙醛脱氢酶2(ALDH2)Glu504Lys基因分型检测技术。方法 以口腔拭子为样本类型,利用ARMS-PCR技术,针对ALDH2 Glu504Lys基因设计特异性引物,在实验室PCR常规反应条件上以金磁纳米层析试纸条为检测平台对各项条件进行优化,以确定最优PCR-金磁微粒ALDH2Glu504Lys口腔拭子反应体系和PCR反应程序。收集60例口腔拭子样本,利用金磁纳米层析检测技术进行检测,并用测序结果进行验证。结果检测的60例口腔拭子样本中杂合突变型16例,野生型42例,纯合突变型2例,金磁微粒层析法检测基因分型结果与测序结果符合率100%。结论 建立了基于金磁纳米微粒ALDH2 Glu504Lys口腔拭子基因检测技术,该方法具有快速、简便和准确的特点,迎合了当下POCT的发展趋势,值得临床推广和应用。
Abstract:
Objective To estabolish the ALDH2 Glu504Lys genotyping technique for oral swab sample type based on “PCR-gold magnetic nanoparticle chromatography1”.Methods According to the ALDH2Glu504Lys gene the specific primers were designed by ARMS-PCR technique.Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR.60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography.The results were verified by sequencing.ResultsThe detected 60 cases of oral swab samples contain 16 cases heterozygous mutation,wild type 42 cases,homozygous mutation 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results.Conclusion The Goldmag nanoparticles chromatographicdetection technique of ALDH2 Glu504Lys buccal swabs system was established.With accuracy,fastness,reliability,it is worthy of clinical promotion.

参考文献/References:

[1] Oyama T,Isse T,Kagawa N,et al.Tissue-distribution of aldehyde dehydrogenase 2 and effects of the ALDH2 gene-disruption on the expression ofenzymes involved in alcohol metabolism[J].Frontiers in Bioscience,2005,10(3):951-960.
[2] Stewart MJ,Malek K,Crabb DW.Distribution of me-ssenger RNAs for aldehyde dehydrogenase 1,aldehyde dehydrogenase 2,and aldehyde dehydrogenase 5 in human tissues[J].Journal of Investigative Medicine,1996,44(2):42-46.
[3] Yoshida A,Huang IY,Ikawa M.Molecular abnormality of an inactive aldehyde dehydrogenase variant commonly found in Orientals[J].Proceedings of the National Academy of Sciences of the United States of America,1984,81(1):258-61.
[4] Chen CH,Ferreira JC,Gross ER,et al.Targeting aldehyde dehydrogenase 2:new therapeutic opportunities[J].Physiological Reviews,2014,94(1):1-34.
[5] Bian Y,Chen YG,Xu F,et al.The polymorphism in aldehyde dehydrogenase-2 gene is associated with elevated plasma levels of high-sensitivity C-reactive protein in the early phase of myocardial infarction[J].Tohoku Journal ofExperimental Medicine,2010,221(2):107-112.
[6] Chang YC,Chiu YF,Lee IT,et al.Common ALDH2,genetic variants predictdevelopment of hypertension in the SAPPHIRe prospective cohort:Gene-environmental interaction with alcohol consumption[J].BMC Cardiovascular Disorders,2012,12(1):58.
[7] Xu F,Chen Y,L(¨overu)R,et al.ALDH2 genetic polymorphism and the risk of type II diabetes mellitus in CAD patients[J].Hypertension Research,2010,33(1):49-55.
[8] Li Y,Zhang D,Jin W,et al.Mitochondrial aldehyde dehydrogenase-2(ALDH2)Glu504Lys polymorphism contributes to the variation in efficacy of sublingual nitroglycerin[J].Journal of Clinical Investigation,2006,116(2):506-511.
[9] 魏 华.金磁微粒层析法结合直扩ARMS-PCR对MTHFR C677T基因多态性检测方法的建立及其临床应用[D].西安:西北大学,2016. Wei Hua.Genotying MTHFR C677T by direct AR-MS-PCR combine with golg-magnetic nanoparticle based on lateral flow assay and its application[D].Xi'an:Northwest University,2016.
[10] Newton CR,Graham A,Heptinstall LE,et al.Analysis of any point mutation in DNA.The amplification refractory mutation system(ARMS)[J].Nucleic Acids Research,1989,17(7):2503-2516.
[11] 李董姣.人乳头瘤病毒PCR-金磁微粒层析快速检测方法的建立[D].西安:西北大学,2015. Li DJ.A rapid detection method of human papillomavirus by PCR-Golg magnetic nanoparticles-based on lateral flow immunossay strips[D].Xi'an:Northwest University,2015.
[12] 张 越,梅善宗.微量口腔粘膜脱落细胞检材STR位点的法医学检测分型[J].皖南医学院学报,2005,24(1):74-76. Zhang Y,Mei SZ.Forensic detection of STR locus in exfoliated cells of micro-oral mucosa[J].Acta Academiae Medicine Wannan,2005,24(1):74-76.
[13] 陈嘉昌,张红宇,彭瑾瑜,等.从唾液获取人体DNA的简易方法与应用[J].中山大学学报(医学科学版),2006,27(S2):171-173. Chen JC,Zhang HY,Peng JY,et al.Simple method and application of obtaining human DNA from saliva[J].Journal of Sun Yat-sen University(Medical Sciences),2006,27(S2):171-173.
[14] 黄娅琳.利用唾液斑DNA上12S rRNA基因片段进行动物种属鉴定[J].中国司法鉴定,2010(5):43-44,48. Huang YL.Animal classification using 12S rRNA genes from salivary stain DNA[J].Chinese Journal of Forensic Sciences,2010(5):43-44,48.
[15] 杨泽民,林 静,陈龙辉,等.一种从唾液中快速提取基因组DNA的方法[J].第二军医大学学报,2015,36(5):548-553. Yang ZM,Lin J,Chen LH,et al.A method for fast genomic DNA extractionfrom saliva and its application[J].Academic Journal of Second Military Medical University,2015,36(5):548-553.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金青年项目(31200749),陕西省教育厅项目(15JK1702)。 作者简介:吕晓玥(1991-),女,研究生在读,临床检验诊断学,E-mail:joymoon@163.com。 通讯作者:刘玉泉,男,E-mail:akzylyq@163.com。
更新日期/Last Update: 1900-01-01