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</div><div>　　定性測定結果確定的依據在于陽性/陰性判定值（cut-off）的建立，cut-off值的確立應盡可能地避免假陽性或假陰性結果的出現。除了cut-off值以外，還有許多其他的設值方法以判斷陰性和陽性結果，如S/N（常稱為P/N）比值（ratio）等。</div><div> </div><div>　　定量測定的依據是使用系列濃度標準品測得的劑量反應曲線（即通常所謂的標準曲線）。由于定量免疫測定中的劑量反應曲線與生化測定中的標準曲線相比，一般均為非線性的，變異較大，故不宜稱為標準曲線。</div><div> </div><div>　　人降鈣素(CT)elisa試劑盒的操作流程：</div><div>　　1、從室溫平衡20min后的鋁箔袋中取出所需板條，剩余板條用自封袋密封放回4℃。</div><div>　　2、設置標準品孔和樣本孔，標準品孔各加不同濃度的標準品50μL；</div><div>　　3、樣本孔中加入待測樣本50μL；空白孔不加。</div><div>　　4、除空白孔外，標準品孔和樣本孔中每孔加入辣根過氧化物酶（HRP）標記的檢測抗體100μL，用封板膜封住反應孔，37℃水浴鍋或恒溫箱溫育60min。</div><div>　　5、棄去液體，吸水紙上拍干，每孔加滿洗滌液（350μL），靜置1min，甩去洗滌液，吸水紙上拍干，如此重復洗板5次（也可用洗板機洗板）。</div><div>　　6、每孔加入底物A、B各50μL，37℃避光孵育15min。</div><div>　　7、每孔加入終止液50μL，15min內，在450nm波長處測定各孔的OD值。</div><div>　　</div><div>　　注意事項：</div><div>　　1、嚴格按照規定的時間和溫度進行溫育以保證準確結果。所有試劑都必須在使用前達到室溫20-25℃。使用后立即冷藏保存試劑；</div><div>　　2、洗板不正確可以導致不準確的結果。在加入底物前確保盡量吸干孔內液體。溫育過程中不要讓微孔干燥掉；</div><div>　　3、消除板底殘留的液體和手指印，否則影響OD值；</div><div>　　4、底物顯色液應呈無色或很淺的顏色，已經變藍的底物液不能使用；</div><div>　　5、封板膜只限一次性使用，避免試劑和標本的交叉污染以免造成錯誤結果；</div><div>　　6、在儲存和溫育時避免強光直接照射。底物請避光保存；</div><div>　　7、平衡至室溫后再打開密封袋以防水滴凝聚在冷板條上。</div> </div> </div> <div class="artview_prev_next"> <p class="artview_prev"><span></span>上一篇：<a href="/News-1458371.html">分享！實驗室用人elisa試劑盒的操作方法和注意事項</a></p> <p 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