GeneXpert技术及原理

GeneXpert技术及原理马林Feb2016真正的开始2©Cepheid–ProprietaryandConfidential重新定义分子诊断TheGeneXpertFamilyofSystemsGeneXpertModuleGeneXpert®CartridgeSmartCyclerSmartCycler®TubeSystemICORE®ModuleAnyTestAnyTimeAnySampleAnyPlace3©Cepheid–ProprietaryandConfidential分子诊断检测（PCR）包含以下步骤：样本采集仪器平台,耗材&软件样本处理扩增检测报告结果SmartCyclerSystem1©Cepheid–Proprietary&ConfidentialGeneXpertSystem©Cepheid–ProprietaryandConfidentialSmartCyclerSmartCyclerII6©Cepheid–ProprietaryandConfidentialSmartCycler反应管25l•不会损坏！7©Cepheid–ProprietaryandConfidential最优化的设计:•快速温度响应•最佳的荧光信号•预防污染一次性反应管LargeSurfaceTemperatureAreawiththinwallenclosureSealablepressurecapPolishedOpticwindows(with90°opticspath)OpticReflectors8©Cepheid–ProprietaryandConfidential固定的组件PAGE12|©Cepheid–ProprietaryandConfidential12BioRadQIAGENRoche系统模块任意配置MultipleI-COREModulesAsSystemBuildingBlocks11©Cepheid–ProprietaryandConfidentialSmartEBVFCTC灵活性:荧光及模块SmartCMVFCTCASRHSVFATAASREVFATA12©Cepheid–ProprietaryandConfidentialMasterMixforaPCRassay•Buffer•MgCl2•TaqPolymerase•dNTPmixture•Primers•Probe•ICPCR反应13©Cepheid–ProprietaryandConfidential扩增/检测SmartMixTMHM预混珠14©Cepheid–ProprietaryandConfidentialReconstitutewithWaterAddthesampleStarttheRun预混珠形式避免加样误差15©Cepheid–ProprietaryandConfidential诊断软件:DX3.03AssayTypesResearchReferenceIVD16©Cepheid–ProprietaryandConfidential©Cepheid–ProprietaryandConfidentialGeneXpert系统GeneXpert技术18©Cepheid–ProprietaryandConfidential整合的微流体技术，包括：•仪器平台•试剂盒•运行程序和对照•实时荧光定量PCR(扩增和检测同时进行)•仪器和样品盒之间不存在湿式接触界面，以消除污染•反应体系完全内部质控-不需要设置独立的外部阳性和阴性对照•集成的细胞超声裂解释放DNA的装置•GeneXpert软件指示单个模块主板以协调阀驱动和整体液压传动•精巧的流体学技术液体流动由微型阀控制适于试剂成分的微量反应•全自动的数据分析和结果解读19©Cepheid–ProprietaryandConfidentialGeneXpert设计赛沛的技术洗涤DNAorRNA提取扩增检测分析阳性阴性DNAorRNA纯化患者样本PCR反应体系SmartCyclerGeneXpert20©Cepheid–ProprietaryandConfidential赛沛的技术SmartCyclerGeneXpert21©Cepheid–ProprietaryandConfidentialI-Core模块模块化设计的SmartCycler和GeneXpert22©Cepheid–ProprietaryandConfidentialGeneXpert6色荧光通道23©Cepheid–ProprietaryandConfidential通道激发波长范围接收波长范围荧光染料1375-405nm420-480nmDye12450-495nm510-535nmDye23500-550nm565-590nmDye34555-590nm606-650nmDye45630-650nm665-685nmDye56630-650nm700nmDye6仪器平台I-CORE模块SmartCycler®GeneXpert®GeneXpert模块BDS24©Cepheid–ProprietaryandConfidentialGeneXpert模块30©Cepheid–ProprietaryandConfidential柱塞杆马达样品盒装载仓室I-CORE主板阀驱动马达超声变幅器GeneXpert模块–试剂盒仓室26©Cepheid–ProprietaryandConfidential阀驱动超声装置注射驱动I-COREGeneXpert试剂盒27©Cepheid–ProprietaryandConfidentialGeneXpert试剂盒28©Cepheid–ProprietaryandConfidentialGeneXpert试剂盒基本特点•GX试剂盒有11个腔体•序号小的腔体用作样本处理•序号大的腔体用于试剂储存•试剂盒类型大致分为两类–CartridgeA–CartridgeC•CartridgeA和C有同样的主体。唯一的不同在于底部构成。29©Cepheid–ProprietaryandConfidential腔体通道ReactionTubeChamberPortSyringeBarrel每个腔体的底部有通道用于腔体流动30©Cepheid–ProprietaryandConfidential试剂盒侧面图示31©Cepheid–ProprietaryandConfidential试剂盒分解图示独特的灵活性:从低到高的通量‘MolecularServerCabinet’系统上使得系统可随检测通量的增长而增长CellChromosomeGenesGenesSequencemRNATranscriptionProteins4646©C©eCpehpehiedid–ProprietaryandConfidential满足多样化测试需求的能力妇女健康危重感染院内感染免疫缺陷性疾病肿瘤遗传性疾病RocheGenProbeQiagenBDAbbottCepheid47©Cepheid–ProprietaryandConfidentialInfinity:更进一步•时间–时间的重要性oInfinity是市场上自动化程度最高的系统•加载试剂盒，Infinity会自动完成剩余工作•更多模块容量,更大的队列载量–无需等待以加载试剂盒o可连接LIS系统•无需人工获取数据•没有不必要的延迟•空间–实验室所急缺o单位面积可获得的最高测试通量oInfinity-80(270cm)/Infinity-48s(216cm)49©Cepheid–ProprietaryandConfidential试剂盒准备最简单的：最复杂的：无需实验室技术的特殊培训52©Cepheid–ProprietaryandConfidential软件步骤点击«CreateTest»53©Cepheid–ProprietaryandConfidential扫描试剂盒二维码54©Cepheid–ProprietaryandConfidential全自动的工作流程•整合了裂解、提取、扩增和实时检测的过程快速获得结果•整合的超声裂解装置以快速获得核酸•特殊的PCR反应管设计可大大缩短PCR循环时间•自动结果解读即时检测•无需批次检测•随时进行任意检测即插即用•无需日常的设置和维护通过整合LIS系统即时获得结果GeneXpert®设计55©Cepheid–ProprietaryandConfidential支持多种样本类型•先进的微流体技术和样本处理可靠和可重复的流程•高度自动化意味着不同用户和地点之间结果的一致性结果的可信性•系统整合完全的内部质控•无需额外的外部对照及阴性对照GeneXpert®设计56©Cepheid–ProprietaryandConfidential污染可能性降至最低•真正的封闭系统，避免样本残留•仪器与试剂盒间无湿式接触界面，消除了交叉污染的可能性优化的故障处理•模块化设计•更少的电子器件，最少的可移动组件•快速的模块校准流程GeneXpert®设计57©Cepheid–ProprietaryandConfidential