单核苷酸多态性位点分析 基质辅助激光解吸电离飞行时间质谱法 编制说明

3本标准根据国标委公布的2024年第一批国家标准计划项目（国标委发[2024]16本标准由全国生化检测标准化技术委员会（SAC/TC387）提出并归口。“MALDI-TOF”）是一种专门用于分析生物大分子的质谱技术。其离子源（MALDI）因使质谱技术能够分析生物大分子而荣获2002年诺贝尔化学奖。此外，MALDI-TOF还可作为多重PCR的质谱检测器，用于单核苷酸多态性位点的分析。酸分析。因此，它在基因分析科学研究、农业、畜牧业疾病控制领域中的病虫害遗传分析、进出口货物鉴定、MALDI-TOF技术在单核苷酸多态性位点分析领域的应用已经历了近二十年的发展。由于我国在核心技术（包括质谱仪、分析软件、试剂等）方面与发近年来，我国在MALDI-TOF质谱仪的研发和制造方面取得了显著进步。目前，国内已有超过十家企业涉足MALDI-TOF质谱仪的研发、生产和销售。其中，一些企业已经开发出目前，我国已有一项行业标准和两项团体标准涉及MALDI-TOF单核苷酸多态性位点分析的相关内容。然而，这些标准反映出当前MALDI-TOF单核苷酸多态性位点分析技术存在的某些技术局限性，以及国外主导企业的专利技术4作为生物遗传物质分析领域的核心技术，MALDI-TOF单核苷酸多态性位点分析技术是我国企业通过自主研发所掌握的少数核心技术之智生物科技（青岛）有限公司、山东英盛生物技术公司等在内的多家企业，正积极致力于MALDI-TOF单核苷酸多态性位点分析的标准化工的标准立项建议。这一举措旨在制定一个更具包容企业的专利技术制约，从而构建一个由我国企业和科研机构主导的MALDI-TOF单核苷酸点分析基质辅助激光解吸电离飞行时间质谱法》标准项目进行了预研，课题组成员广泛收集了国内外近百余篇标准、文献，了解了国内外相关技（3）2024年4月收到全国生化检测标准化技术委员会生化标[2024]5文件《关于下达2024年第一批推荐性国家标准计划的通知文件》以及该标委会转发的国标委发[2024]16号《国家标准委关于下达2024年第一批国家标准制计划及相核苷酸多态性位点分析基质辅助激光解吸电离飞行时间质谱法论，提出了宝贵的意见和建议。标准起草小组根据专家吸电离飞行时间质谱法》标准的起草研制工作。完成了《单核苷酸辅助激光解吸电离飞行时间质谱法》标准的编制说明，会汇报了《单核苷酸多态性位点分析基质辅助激光解吸电离飞行案）进一步完善和修改情况，向全国生化检测标准化技位点分析基质辅助激光解吸电离飞行时间质谱法》标准（征求并参照GB/T6379.1-2004《测量方法与结果的准确度（正确度与精密度）第1部分总则与定义》和GB/T6379.2-2004《测量方法与结果的准确度（正确度与精密度）第2部分确定2、确定国家标准主要内容（如技术指标、参数、长度8~25）bp；熔解温度（Tm55~65）℃;表1PCR反应体系组成成分浓度/体积说明DNA模板（1~100）ng来源：全血、组织、唾液等PCR预混液含Taq酶、MgCl₂、dNTP混合液引物（正/反）每种引物在反应液中的终浓度为（0.1~0.2）μM无RNA酶水补足至5μL/每对引物扩增的单重扩增产物电泳检测为单一条带；琼脂糖凝胶电泳（2%）。产物浓度≥5ng/μL；使用UV分光光度计或荧光仪（如Qubit）检测。表2SAP反应体系组成成分浓度/体积说明PCR产物来自前述PCR反应体系SAP缓冲液（0.17~1）μL厂商提供（含Mg²⁺)SAP酶（0.5~1）U依据厂商推荐调整用量无RNA酶水补足至7μL/）；表3MPE延伸反应体系组成成分浓度/体积说明SAP处理后的PCR产物上一步产物延伸缓冲液配合延伸酶使用，根据厂商提供的参考用量MPE引物特异性延伸引物延伸酶（如ThermoSequenase）0.6μL热稳定聚合酶，根据厂商提供的参考用量混合ddNTP含不同分子质量标记，保证峰分辨）：表4纯化体系成分浓度/体积说明MPE反应产物来自前一步延伸反应纯化树脂（3~10）mg（固体）每次实验根据厂商建议称取去离子水用于稀释反应体系及助于均匀分散树脂样品与树脂混匀，震荡（或颠倒混匀10~30）min；）；表5基质配制与添加方法步骤操作说明1配制3-HPA溶液，过滤除杂（0.22μm）2将延伸产物以1:1体积比与基质混合3点样：每孔加1μL基质，干燥后加1μL基质产物样品混合液4室温自然干燥结晶（优于PAGE纯化级别）、SAP酶、SAP缓冲液、单碱基延伸酶（MPE酶）、单碱基延阴性质控：应有标准物质溯源信息或经测序确认；阳性质控：应有标准物质溯源信息MALDI-TOFMS：质谱仪的质量范围须满足最小值≤1500，最大值≥10000；质量分所有样本均视为有潜在的感染性，操作及运输提取的模板DNA纯度要求紫外分光光度计读数（A260/A280）在1.7~将PCR试剂取出，在冰上融解，振荡混匀（3~5）s，瞬时离心。根据检测样本数目，按照表6的比例在适合的离心管中配制PCR扩增反应混合液（DNA模板除外），然后将试配制好的PCR反应体系涡旋振荡混匀，瞬时离心，按3μL/孔的量加入PPCR反应管底部。每次反应选择两个孔分别加入2μL阴表6PCR反应体系试剂名称反应孔/μL成分及反应液浓度PCR预混液2.0dNTP混合液（0.1μM）无酶水补充体系/PCR引物DNA模板2.0总体积5.0/表7PCR反应程序扩增温度95℃95℃59℃72℃60℃4℃反应时间15min30s30s15minHold备注45cycles将SAP酶和SAP缓冲液平衡至室温，振荡混匀（3~5）s，瞬时离心。根据检测样本数目，按照表8的比例在适合的离心管中配制PCR扩增反应混合液（DNA模板除外），上述表8SAP反应体系试剂名称反应孔/μL无酶水SAP缓冲液（10×)0.17SAP酶（1.7U/μL）0.3总体积2.0将SAP反应管板放置于PCR仪中，运行表9表9SAP反应程序扩增温度反应时间37℃40min5min4℃Hold将MPE酶和缓冲液、ddNTP混合液和MPE引物平例在适合的离心管中配制MPE反应混合液，混匀后按4μL每SAP反应孔分装。上述操作须表10MPE反应体系试剂名称反应孔/μLddNTP混合液MPE酶缓冲液MPE酶（1.5U/μL）0.6MPE引物（2~5μM）总体积4.0将MPE反应管板放置于PCR仪中，运行表1表11MPE反应程序扩增温度95℃95℃48℃72℃4℃反应时间30sHold备注40cycles5cycles将PCR试剂取出，在冰上融解，振荡混匀（3~5）s，瞬时离心。根据检测样本数目，），配制好的PCR反应体系涡旋振荡混匀，瞬时离心，按3μL/孔的量加入PPCR反应管底部。每次反应选择两个孔分别加入2μL阴表12PCR反应体系试剂名称反应孔/μL成分及反应液浓度PCR预混液2.0混合液（0.1μM）无酶水补充体系/PCR引物DNA模板1.0~2.0总体积5.0/将PCR反应管放置于PCR仪中，运行表表13PCR反应程序扩增温度95℃95℃60℃72℃60℃4℃反应时间15min30s30s15minHold备注45cycles将SAP酶和SAP缓冲液平衡至室温，振荡混匀（3~5）s，瞬时离心。根据检测样本数表14SAP反应体系试剂名称反应孔/μL无酶水SAP缓冲液（10×)0.17SAP酶（1.7U/μL）0.3总体积2.0将SAP反应管板放置于PCR仪中，运行表15表15SAP反应程序扩增温度反应时间37℃40min5min4℃Hold将MPE酶和缓冲液、ddNTP混合液和MPE引物平例在适合的离心管中配制MPE反应混合液，混匀后按4μL每SAP反应孔分装。上述操作须表16MPE反应体系试剂名称反应孔/μLddNTP混合液MPE酶缓冲液MPE酶（1.5U/μL）0.6MPE引物（2~5μM）总体积4.0将MPE反应管板放置于PCR仪中，运行表1表17MPE反应程序扩增温度94℃94℃58℃72℃4℃反应时间30sHold备注40cycles5cycles将PCR试剂取出，在冰上融解，振荡混匀（3~5）s，瞬时离心。根据检测样本数目，），配制好的PCR反应体系涡旋振荡混匀，瞬时离心，按3μL/孔的量加入PPCR反应管底部。每次反应选择两个孔分别加入2μL阴表18PCR反应体系试剂名称反应孔/μL成分及反应液浓度PCR预混液2.0混合液（0.1μM）无酶水补充体系/PCR引物DNA模板2总体积5.0/将PCR反应管放置于PCR仪中，运行表扩增温度95℃95℃65℃72℃60℃4℃表19PCR反应程序反应时间15min30s30s15minHold备注45cycles将SAP酶和SAP缓冲液平衡至室温，振荡混匀（3~5）s，瞬时离心。根据检测样本数），表20SAP反应体系试剂名称反应孔/μL无酶水SAP缓冲液（10×)0.17SAP酶（1.7U/μL）0.3总体积2.0将SAP反应管板放置于PCR仪中，运行表21表21SAP反应程序扩增温度反应时间37℃40min5min4℃Hold将MPE酶和缓冲液、ddNTP混合液和MPE引物平例在适合的离心管中配制MPE反应混合液，混匀后按4μL每SAP反应孔分装。上述操作须表22MPE反应体系试剂名称反应孔/μLddNTP混合液MPE酶缓冲液MPE酶（1.5U/μL）0.6MPE引物（2~5μM）总体积4.0将MPE反应管板放置于PCR仪中，运行表2表23MPE反应程序扩增温度95℃95℃52℃72℃4℃反应时间30sHold备注40cycles5cycles将校准品和无核酸酶水按照体积比1:13的比例，配制校准品混合液。在每个反应孔中样品的每个孔对齐。然后轻敲树脂管，使树脂落入样本树脂的样品板放置在翻转混匀仪中，以20rpm的速率混匀30min。混匀结束后瞬时离心，将纯化后的产物与基质混合，取适量体积点在靶板上（通过质谱软件，分析基因位点的形态。应能够给出检测样本中SNP位点的定性分析结表24耳聋判定结果表突变所在基因突变所在位点描述野生(阴性）突变（阳性）杂合GJB2c.35delGGDELG/DELc.257C>GCGC/G突变所在基因突变所在位点描述野生(阴性）突变（阳性）杂合c.427C>TCTC/Tc.167delTTDELT/DELc.176_191del16ACACGTTCTTGCAGCCDELACACGTTCTTGCAGCC/DELc.235delCCDELC/DELc.299_300delATATDELAT/DELc.512insAACGDELAACGDEL/AACGSLC26A4c.281C>TCTC/Tc.589G>AGAG/Ac.1174A>TATA/Tc.1226G>AGAG/Ac.1229C>TCTC/Tc.1975G>CGCG/Cc.2027T>ATAT/Ac.2162C>TCTC/Tc.2168A>GAGA/GIVS7-2A>GAGA/GIVS15+5G>AGAG/A12srRNAm.1494C>TCTC/Tm.1555A>GAGA/GGJB3c.538C>TCTC/Tc.547G>AGAG/AUSH2A2314delGGDELG/DEL表25地中海贫血判定结果表突变所在基因突变所在位点描述野生(阴性）突变（阳性）杂合α-珠蛋白基因c.369C>GCGC/Gc.427T>CTCT/Cc.377T>CTCT/Cβ-珠蛋白基因c.45dupGGdupGG/dupGc.52A>TATA/Tc.85dupCCdupCC/dupCc.94delCCDELC/DELc.126_129delCTTTCTTTDELCTTT/DELc.130G>TGTG/Tc.217dupAAdupAA/dupAc.2T>GTGT/Gc.-82C>ACAC/Ac.-80T>CTCT/Cc.-79A>GAGA/G突变所在基因突变所在位点描述野生(阴性）突变（阳性）杂合c.-78A>GAGA/Gc.-50A>CACA/Cc.92+1G>TGTG/Tc.316-197C>TCTC/Tc.92+5G>CGCG/Cc.79G>AGAG/A表25心血管指导用药判定结果表突变所在基因突变所在位点描述野生(阴性）突变（阳性）杂合ADRB1rs1801253,G>CGCG/CAGTR1rs5186,A>CACA/CALDH2rs671,G>AGAG/AAPOErs7412,C>TCTC/Trs429358,T>CTCT/CCYP2C19rs12248560,C>TCTC/Trs4244285,G>AGAG/Ars4986893,G>AGAG/ACYP2C9rs1799853,C>TCTC/Trs1057910,A>CACA/CCYP2D6rs1065852,G>AGAG/ASLCO1B1rs4149056,T>CTCT/Crs11045852,A>GAGA/GVKORC1rs9923231,C>TCTC/Trs9934438,G>AGAG/A实验方法：采用梯度稀释法对检测限样本进行验证，将阴阳质控品、样品稀释至10判定标准：在10ng/μL、5ng/μL的浓度下，所有样本的所有位点均应被均应被检出，且检出的分型结果与理论位点及分型表26耳聋阳性质控品(10ng/μL)SNP分析结果123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCCTTTTTTTTTT6CCCCCCCCCCc.299_300delAT4CCCCCCCCCCGGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGTTTTTTTTTTCCCCCCCCCCAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGUSH2AGGGGGGGGGG表27耳聋阳性质控品(5ng/μL)SNP分析结果123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCCTTTTTTTTTT6123456789CCCCCCCCCCc.299_300delATc.512insAACG4CCCCCCCCCCGGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGTTTTTTTTTTCCCCCCCCCCAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGUSH2AGGGGGGGGGG表28耳聋阳性质控品(2.5ng/μL)SNP分析结果123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCCTTTTTTTTTT6CCCCCCCCCCc.299_300delATc.512insAACG4CCCCCCCCCCGGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCC123456789GGGGGGGGGGTTTTTTTTTTCCCCCCCCCCAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGUSH2AGGGGGGGGGG表29耳聋阴性质控品(10ng/μL)SNP分析结果123456789GGGGGGGGGGTTTTTTTTTTc.176_191del16c.235delCc.299_300delATc.512insAACG4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2A表30耳聋阴性质控品(5ng/μL)SNP分析结果123456789GGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2A表31耳聋阴性质控品(2.5ng/μL)SNP分析结果123456789GGGGGGGGGGTTTTTTTTTT123456789c.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2A表32地中海贫血阳性质控品(10ng/μL)SNP分析结果地中海贫血阳性质控品(10ng/μL)123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCC地中海贫血阳性质控品(10ng/μL)123456789GGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTCCCCCCCCCCAAAAAAAAAA表33地中海贫血阳性质控品(5ng/μL)SNP分析结果123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTCCCCCCCCCCAAAAAAAAAA表34地中海贫血阳性质控品(2.5ng/μL)SNP分析结果地中海贫血阳性质控品(2.5ng/μ123456789GGGGGGGGGGCCCCCCCCCCCCCCCCCCCC地中海贫血阳性质控品(2.5ng/μ123456789TTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTCCCCCCCCCCAAAAAAAAAA表35地中海贫血阴性质控品(10ng/μL)SNP分析结果地中海贫血阴性质控品(10ng/μL)123456789CCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAA地中海贫血阴性质控品(10ng/μL)123456789AAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGG表36地中海贫血阴性质控品(5ng/μL)SNP分析结果123456789CCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGG表37地中海贫血阴性质控品(2.5ng/μL)SNP分析结果地中海贫血阴性质控品(2.5ng/μ123456789CCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAA地中海贫血阴性质控品(2.5ng/μ123456789CCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGG表38心血管指导用药阳性质控品(10ng/μL)SNP分析结果123456789CCCCCCCCCCCCCCCCCCCCAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGTTTTTTTTTTAAAAAAAAAA表39心血管指导用药阳性质控品(5ng/μL)SNP分析结果123456789123456789CCCCCCCCCCCCCCCCCCCCAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGTTTTTTTTTTAAAAAAAAAA表40心血管指导用药阳性质控品(2.5ng/μL)SNP分析结果123456789CCCCCCCCCCCCCCCCCCCCAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAACCCCCCCCCCGGGGGGGGGGTTTTTTTTTTAAAAAAAAAA表41心血管指导用药阴性质控品(10ng/μL)SNP分析结果123456789123456789GGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCAAAAAAAAAAGGGGGGGGGGTTTTTTTTTTAAAAAAAAAACCCCCCCCCCGGGGGGGGGG表42心血管指导用药阴性质控品(5ng/μL)SNP分析结果123456789GGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCAAAAAAAAAAGGGGGGGGGGTTTTTTTTTTAAAAAAAAAACCCCCCCCCCGGGGGGGGGG表43心血管指导用药阴性质控品(2.5ng/μL)SNP分析结果123456789123456789GGGGGGGGGGAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCAAAAAAAAAAGGGGGGGGGGTTTTTTTTTTAAAAAAAAAACCCCCCCCCCGGGGGGGGGG判定标准：阴阳质控品的所有样本的所有位点均从表44~52可以看出，阴阳质控品的所有样本的所有位点均应被检出，且检出的分型结果与理论位点及分型信息完全一致，样品的7次测定结果一致，表明重复性与方法准确表44耳聋阳性质控品SNP分析结果述耳聋阳性质控品SNP分析结果1234567GGGGGGGCCCCCCCCCCCCCCTTTTTTTCCCCCCC述耳聋阳性质控品SNP分析结果1234567CCCCCCCGGGGGGGAAAAAAAGGGGGGGCCCCCCCGGGGGGGTTTTTTTCCCCCCCAAAAAAAAAAAAAAGGGGGGGCCCCCCCAAAAAAACCCCCCCGGGGGGGUSH2AGGGGGGG表45耳聋阴性质控品SNP分析结果述耳聋阴性质控品SNP分析结果1234567GGGGGGGTTTTTTTTTTTTTTAAAAAAATTTTTTTAAAAAAATTTTTTTCCCCCCCAAAAAAATTTTTTTGGGGGGGGGGGGGG述耳聋阴性质控品SNP分析结果1234567AAAAAAATTTTTTTGGGGGGGTTTTTTTAAAAAAAUSH2A表46样品耳聋SNP分析结果述样品SNP分析结果1234567GGGGGGGTTTTTTTTTTTTTTAAAAAAATTTTTTTAAAAAAATTTTTTTCCCCCCCAAAAAAATTTTTTTGGGGGGGGGGGGGGAAAAAAATTTTTTTGGGGGGGTTTTTTTAAAAAAAUSH2A表47地中海贫血阳性质控品SNP分析结果12345671234567GGGGGGGCCCCCCCCCCCCCCTTTTTTTTTTTTTTGGGGGGGAAAAAAACCCCCCCGGGGGGGGGGGGGGCCCCCCCTTTTTTTTTTTTTTCCCCCCCAAAAAAA表48地中海贫血阴性质控品SNP分析结果CCCCCCCTTTTTTTTTTTTTTGGGGGGGAAAAAAACCCCCCCCCCCCCCGGGGGGGAAAAAAATTTTTTTCCCCCCCTTTTTTTAAAAAAAAAAAAAACCCCCCCAAAAAAAGGGGGGGCCCCCCCGGGGGGGGGGGGGGCCCCCCC表49地中海贫血样品SNP分析结果CCCCCCCTTTTTTTTTTTTTTGGGGGGGAAAAAAACCCCCCCCCCCCCCGGGGGGGAAAAAAATTTTTTTCCCCCCCTTTTTTTAAAAAAAAAAAAAAAAAAAAAGGGGGGGCCCCCCCGGGGGGGGGGGGGGCCCCCCC表50心血管指导用药阳性质控品SNP分析结果1234567CCCCCCCCCCCCCCAAAAAAATTTTTTT1234567CCCCCCCTTTTTTT9AAAAAAAAAAAAAATTTTTTTCCCCCCCAAAAAAACCCCCCC1GGGGGGGTTTTTTTAAAAAAA表51心血管指导用药阴性质控品SNP分析结果1234567CCCCCCCCCCCCCCAAAAAAATTTTTTTCCCCCCCTTTTTTTAAAAAAAAAAAAAATTTTTTTCCCCCCCAAAAAAACCCCCCCGGGGGGGCCCCCCCCCCCCCC表52心血管指导用药样品SNP分析结果1234567G,AG,AG,AG,AG,AG,AG,ACCCCCCCAAAAAAAGGGGGGG1234567CCCCCCCTTTTTTTCCCCCCCGGGGGGGGGGGGGGCCCCCCCAAAAAAAG,AG,AG,AG,AG,AG,AG,ATTTTTTTAAAAAAAC,TC,TC,TC,TC,TC,TC,T收集了50管心血管疾病病人的全血，采用天根生化科技（北京）有限公司（磁珠法通用型基因组DNA提取试剂盒,货号：DP705）。按试剂盒中提供的操作说明书提取样本表53耳聋50个样本SNP分析结果123456789GGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2AGGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2AGGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2AGGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2AGGGGGGGGGGTTTTTTTTTTc.299_300delAT4TTTTTTTTTTAAAAAAAAAATTTTTTTTTTAAAAAAAAAATTTTTTTTTTCCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGGGGGGGGGGGGGGGAAAAAAAAAATTTTTTTTTTGGGGGGGGGGTTTTTTTTTTAAAAAAAAAAUSH2A表54地中海贫血50个样本SNP分析结果123456789CCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCCCCCCCCCCCGGGGGGGGGGAAAAAAAAAATTTTTTTTTTCCCCCCCCCCTTTTTTTTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGGGGGGGCCCCCCCCCCGGGGGGGGGGGGGGGGGGGGCCCCCCCCCCTTTTTTTTTTTTTTTTTTTTGGGGGGGGGGAAAAAAAAAACCCCCCCCCCC