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<strong><span style="text-decoration-line: underline;"><span style="font-family: 方正姚体; line-height: 24px;">操作注意事项</span></span></strong>
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<span style="font-family: 方正姚体;">1.</span><span style="font-family: 方正姚体; line-height: 24px;"> 试剂盒保存在2-8℃，使用前室温平衡20分钟。从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶完全溶解后再使用。</span>
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<span style="font-family: 方正姚体;">2.</span><span style="font-family: 方正姚体; line-height: 24px;"> 实验中不用的板条应立即放回自封袋中，密封（低温干燥）保存。</span>
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<span style="font-family: 方正姚体;">3.</span><span style="font-family: 方正姚体; line-height: 24px;"> 浓度为0的S0号标准品即可视为阴性对照或者空白；按照说明书操作时样本已经稀释5倍，结果乘以5才是样本实际浓度。</span>
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<span style="font-family: 方正姚体;">4.</span><span style="font-family: 方正姚体; line-height: 24px;"> 严格按照说明书中标明的时间、加液量及顺序进行温育操作。</span>
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<span style="font-family: 方正姚体;">5.</span><span style="font-family: 方正姚体; line-height: 24px;"> 所有液体组分使用前充分摇匀。</span>
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<strong><span style="text-decoration-line: underline;"><span style="font-family: 方正姚体; line-height: 150%;">需自备的物品</span></span></strong>
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<span style="font-family:方正姚体;font-size:16px">1. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"><span style="font-family:方正姚体">酶标仪（</span>450nm）</span>
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<span style="font-family:方正姚体;font-size:16px">2. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"><span style="font-family:方正姚体">高精度加样器及枪头：</span>0.5-10uL、2-20uL、20-200uL、200-1000uL</span>
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<span style="font-family:方正姚体;font-size:16px">3. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px">37℃恒温箱</span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">试剂的准备</span></span></strong>
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<span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">20×洗涤缓冲液的稀释：蒸馏水按1：20稀释，即1份的20×洗涤缓冲液加19份的蒸馏水。</span></span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">洗板方法</span></span></strong>
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<span style="font-family:方正姚体;font-size:16px">1.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">手工洗板：甩尽孔内液体，每孔加满洗涤液，静置</span>1min后甩尽孔内液体，在吸水纸上拍干，如此洗板5次。</span>
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<span style="font-family:方正姚体;font-size:16px">2.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">自动洗板机：每孔注入洗液</span>350μL，浸泡1min，洗板5次。</span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">操作步骤</span></span></strong>
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<span style="font-family:方正姚体;font-size:16px">1.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">从室温平衡</span>20min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。</span>
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<span style="font-family:方正姚体;font-size:16px">2.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">设置标准品孔和样本孔，标准品孔各加不同浓度的标准品</span>50μL；</span>
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<span style="font-family:方正姚体;font-size:16px">3.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">样本孔先加待测样本</span>10μL，再加样本稀释液40μL；空白孔不加。</span>
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<span style="font-family:方正姚体;font-size:16px">4.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">除空白孔外，标准品孔和样本孔中每孔加入辣根过氧化物酶（</span>HRP）标记的检测抗体100μL，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。</span>
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<span style="font-family:方正姚体;font-size:16px">5.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">弃去液体，吸水纸上拍干，每孔加满洗涤液，静置</span>1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。</span>
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<span style="font-family:方正姚体;font-size:16px">6.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">每孔加入底物</span>A、B各50μL，37℃避光孵育15min。</span>
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<span style="font-family:方正姚体;font-size:16px">7.</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">每孔加入终止液</span>50μL，15min内，在450nm波长处测定各孔的OD值。</span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">结果判断</span></span></strong>
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<span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">绘制标准曲线：在</span>Excel工作表中，以标准品浓度作横坐标，对应OD值作纵坐标，绘制出标准品线性回归曲线，按曲线方程计算各样本浓度值。</span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">试剂盒性能</span></span></strong>
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<span style="font-family:方正姚体;font-size:16px">1. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">准确性：标准品线性回归与预期浓度相关系数</span>R值，大于等于0.9900。</span>
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<span style="font-family:方正姚体;font-size:16px">2. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">灵敏度：低检测浓度小于</span>1.0 </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px">ng/ml</span><span style=";font-family:方正姚体;line-height:150%;font-size:16px">。</span>
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<span style="font-family:方正姚体;font-size:16px">3. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">特异性：不与其它可溶性结构类似物交叉反应。</span></span>
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<span style="font-family:方正姚体;font-size:16px">4. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">重复性：板内、板间变异系数均小于</span>15%。</span>
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<span style="font-family:方正姚体;font-size:16px">5. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">贮藏：</span>2-8℃，避光防潮保存。</span>
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<span style="font-family:方正姚体;font-size:16px">6. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">有效期：</span>6个月</span>
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<strong><span style="text-decoration:underline;"><span style="font-family: 方正姚体;line-height: 150%;font-size: 16px">免责声明</span></span></strong>
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<span style="font-family:方正姚体;font-size:16px">1. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">试剂盒仅供研究使用，不得用于临床实验或人体实验，否则所产生的一切后果，由实验者承担，本公司概不负责。</span></span>
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<span style="font-family:方正姚体;font-size:16px">2. </span><span style=";font-family:方正姚体;line-height:150%;font-size:16px"> <span style="font-family:方正姚体">严格按照说明书操作，实验者违反说明书操作，后果由实验者承担。</span></span>
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