1、结 合洗涤洗脱磁粒法提取核酸示意图裂解结合磁分离吸去废液洗涤洗脱磁架提取核酸示意图裂解结合转移磁粒洗涤洗脱转移磁粒磁笔提取核酸示意图 自动提取的原理与磁笔提取相同,只是用机器的运转代替手工操作,实现高通量提取和减少工作量。Ct值方法HCV样本1 HCV样本2 HCV样本3Promega体系平行127.6030.2127.39Promega体系平行227.7230.8427.20Qiagen kit平行127.8639.0028.82Qiagen kit平行227.3333.5828.55方法HBV样本1HBV样本2Promega体系平行126.6532.78Promega体系平行226.843
2、2.52Qiagen kit平行126.7032.96Qiagen kit平行226.7132.28HCV阳性样本10倍稀释梯度:100,10-1,10-2,10-3,10-4磁珠提取与柱提取法比较(数据见下页)稀释组柱提取磁珠提取10-024.1324.3425.2810-127.1529.2028.5710-231.2333.3432.6410-335.3435.8735.3910-438.1638.0738.69磁珠提取方法与柱提取法及水溶液梯度稀释样本比较磁珠提取与柱提法的提取效果相当。在低浓度样本提取方面,荧光增量与稀释组无显著差异,此方法在去除样本中抑制物方面效果良好。10份阳性参
3、考品:P1P10HCV阳性参考品CtHCV阳性参考品CtP134.58P6NDP2NDP736.18P336.96P8NDP434.81P932.16P535.44P1036.73 HIV阳性样本梯度稀释成1/5,1/25,1/50,1/100和1/200五个浓度,提取RNA,原样本提取后用TE按相同比例稀释,上机比较。稀释组磁珠提取1-28.881/532.0931.291/535.7833.931/5037.1735.591/10038.8336.121/200-37.33HIV 样本稀释梯度:1/5,1/25,1/50,1/100和1/200HIV核酸国家参考品检测HIV RNA灵敏度和定量参考品:B2B5(至少检出B2-B3)HIV RNA线性参考品:A2A5。线形相关系数:0.9907HIV 阳性参考品:P1,P2,P3,P5,P7,P8 HIV阳性参考品CtHIV阳性参考品CtP132.94P526.42P235.98P728.93P337.61P832.55HIV阳性参考品检测灵敏度参考品L0L1L2L3L4L5L6理论浓度对数值8.3427.6196.6495.6724.7153.6362.043Ct15.0416.9720.4022.0426.3429.0335.17