全血PCR直接扩增技术在亚甲基四氢叶酸还原酶基因C677T多态性分析中的应用全血PCR直接扩增技术在亚甲基四氢叶酸还原酶基因C677T多态性分析中的应用刘冉1，尹立红1*，浦跃朴1，周国华2，王仪3，潘恩春3，杨文洲3摘要：［目的］应用全血PCR直接扩增技术检测淮安汉族人群亚甲基四氢叶酸还原酶基因MTHFRC677T多态性并分析其地理分布特征，评价该技术在大规模现场人群基因多态性筛检中的应用价值。［方法］应用全血PCR直接扩增技术分析205例淮安汉族健康个体MTHFRC677T的基因型分布频率，并与传统的聚合酶链反应—限制性片段长度多态性（Polymerasechainreactionandrestrictionfragmentlengthpolymorphism，PCR-RFLP）技术检测结果进行比对。并将检测结果与已报道的不同地区汉族人群资料进行比较。［结果］全血PCR直接扩增技术与传统PCR-RFLP法测定MTHFRC677T多态性的分布频率具有高度一致性（Kappa=0.946，95%CI：0.906~0.1015）。结果表明淮安汉族人群MTHFRC677T的基因型频率分别为CC32.68%、CT46.83%、TT20.49%；C和T等位基因频率分别为56.10%和43.90%；结合其他9个地区的汉族人群MTHFR677T等位基因频率的趋势性χ2检验，发现MTHFR677T的分布随地理纬度的降低，T等位基因频率也降低（P=0.001）。［结论］全血PCR直接扩增技术简便易行，适用于人群基因多态性的快速筛检。在所研究的10个中国汉族人群中MTHFRC677T多态性显示与不同地理纬度的地区分布有统计学相关性。关键词：外周血；亚甲基四氢叶酸还原酶；单核苷酸多态；筛检技术MTHFRC677TPolymorphismRapidlyDetectedwithDirectAmpli?cationProtocolinPeripheralBloodSamplesLIURan1，YINLi-hong12*，PUYue-pu1，ZHOUGuo-hua2，WANGYi3，PANEn-chun3，YANGWen-zhou3（1.SchoolofPublicHealth，SoutheastUniversity，JiangsuNanjing210109，China；2.HuadongResearchInstituteforMedicineandBiotechnics，JiangsuNanjing210102，China；3.HuaianCenterforDiseaseControlandPrevention，JiangsuHuaian223001，China）Abstract：［Objective］Toexaminethefrequencydistributionofmethylenetetrahydrofolatereductase（MTHFR）C677TpolymorphismsinHuaianHanpopulationandtoevaluatetheapplicationofDirectAmplificationProtocolfordetectinggeneticpolymorphismsinpopulation.［Methods］PolymorphismsofMTHFRC677Tin205HuaianHanhealthyvolunteerswasdetectedwithDirectAmplificationProtocolandRFLPtechnique，respectively.ThefrequencyofMTHFRC677TgenetypeswascomparedwithotherHanpopulationdatafromdifferentareasinChina.［Results］TherewashighconsistencyofMTHFRC677TpolymorphismsbetweenthemethodwithbloodsampleandwithgenomicDNA（Kappa=0.946，95%CI0.906~0.1015）.ThefrequenciesoftheC/C，C/TandT/Tgenotypesinthesubjectswere32.68%、46.83%and20.49%respectively，andtheallelefrequenciesof677Cand677Twere56.10%and43.90%respectively.TheanalysisofMTHFR677TallelefrequencywithHanpopulationinotherninedifferentareasinChinashowedthedistributionofMTHFR677Twasdecreasedwithgeographicallatitude（P=0.001）.［Conclusion］ThefrequencyofthegeneticpolymorphismsofMTHFRC677TinChineseHanpopulationsfromdifferentareasdependsongeographicallatitude.TheMTHFRC677TdistributionofHuaianHanpopulationgroupisconsistentwiththemodelofpopulationgroupswiththelatituderangebetweenN30°andN35°.DirectAmplificationProtocolinperipheralbloodsamplesisagoodsystemofdirectPCRamplificationwithbloodsampleforrapididentificationofgeneticpolymorphisms.KeyWords：peripheralblood；methylenetetrahydrofolatereductase；singlenucleotidepolymorphism；screening基金项目：国家自然科学基金资助项目（编号：30571540，30671732）作者简介：刘冉（1974-），女，博士生，研究方向：肿瘤环境基因组学*通讯作者（Correspondentauthor）：尹立红教授，E-mail：lhyin@https://www.sodocs.net/doc/5d16659507.h...