實驗時elisa競爭法檢測抗-HBe（快速法）

<p class="MsoNormal"><span style="color: rgb(255, 153, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">實驗原理</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">用抗</font>-HBe<font face="宋體">包被固相載體后，同時加被檢血清和合適滴度中和試劑</font><font face="Calibri">HBeAg</font><font face="宋體">，若被檢血清中含有抗</font><font face="Calibri">-HBe</font><font face="宋體">，則與包被抗</font><font face="Calibri">-HBe</font><font face="宋體">競爭結合</font><font face="Calibri">HBeAg</font><font face="宋體">，被檢標本中抗</font><font face="Calibri">-HBe</font><font face="宋體">越多，</font><font face="Calibri">HBeAg</font><font face="宋體">被結合越多，而與包被抗</font><font face="Calibri">-HBe</font><font face="宋體">結合就越少，加底物后顯色就越淺；反之越深。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 153, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">&nbsp;</span></strong></span></p> <p class="MsoNormal"><span style="color: rgb(255, 153, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">主要試劑與器材</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">elisa<font face="宋體">試劑盒：</font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">內含已包被抗</font><font face="Calibri">-HBe</font><font face="宋體">板條、</font><font face="Calibri">HRP-</font><font face="宋體">抗</font><font face="Calibri">HBe</font><font face="宋體">溶液、中和試劑</font><font face="Calibri">HBeAg</font><font face="宋體">溶液、陽性和陰性對照血清，底物溶液（</font><font face="Calibri">A</font><font face="宋體">液、</font><font face="Calibri">B</font><font face="宋體">液）、終止液、洗滌液。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">結果分析<o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">P/N<font face="宋體">&le;</font><font face="Calibri">0.3</font><font face="宋體">為陽性。</font><font face="Calibri">P/N</font><font face="宋體">＞</font><font face="Calibri">0.3</font><font face="宋體">為陰性。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">P/N=<font face="宋體">待檢血清</font><font face="Calibri">OD</font><font face="宋體">值</font><font face="Calibri">/</font><font face="宋體">陰性對照</font><font face="Calibri">OD</font><font face="宋體">值。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 153, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">注意事項</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">1<font face="宋體">、</font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">試劑置</font><font face="Calibri">2-4</font><font face="宋體">℃保存。取用時應先置室溫平衡。干包被板條須密封防潮，</font><font face="Calibri">zui</font><font face="宋體">好凍存，取用后余者及時封存。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">2<font face="宋體">、</font></span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋體">恰當選好</font><font face="Calibri">HBeAg</font><font face="宋體">的濃度。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">3</span></strong><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><strong><font face="宋體">、</font></strong><font face="宋體">試劑用前應搖勻。</font><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="color: rgb(255, 153, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;">操作步驟</span></strong></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">1<font face="宋體">、加樣：取出干包板條，按編號順序分別加</font><font face="Calibri">50</font><font face="宋體">&mu;</font><font face="Calibri">l</font><font face="宋體">待檢血清、陰性對照血清和陽性對照血清于相應孔中。設空白對照孔，除此孔外，每孔加</font><font face="Calibri">HBeAg50</font><font face="宋體">&mu;</font><font face="Calibri">l</font><font face="宋體">、</font><font face="Calibri">HRP-</font><font face="宋體">抗</font><font face="Calibri">HBe50</font><font face="宋體">&mu;</font><font face="Calibri">l</font><font face="宋體">，振蕩混勻后，置</font><font face="Calibri">43</font><font face="宋體">℃（或</font><font face="Calibri">37</font><font face="宋體">℃）溫育</font><font face="Calibri">1h</font><font face="宋體">。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">2<font face="宋體">、洗滌：甩去孔內液體，用洗滌液注滿各孔，靜置</font><font face="Calibri">20s</font><font face="宋體">，甩干。如此重復洗滌</font><font face="Calibri">4</font><font face="宋體">次，在吸水紙上拍干。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">3<font face="宋體">、顯色：每孔加顯色劑</font><font face="Calibri">A</font><font face="宋體">、顯色劑</font><font face="Calibri">B</font><font face="宋體">各</font><font face="Calibri">50</font><font face="宋體">&mu;</font><font face="Calibri">l</font><font face="宋體">，振蕩混勻后置</font><font face="Calibri">37</font><font face="宋體">℃避光顯色</font><font face="Calibri">10-15min</font><font face="宋體">。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">4<font face="宋體">、終止：每孔加終止液</font><font face="Calibri">50</font><font face="宋體">&mu;</font><font face="Calibri">l</font><font face="宋體">，振蕩混勻。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">5<font face="宋體">、酶標儀檢測：選用</font><font face="Calibri">450nm</font><font face="宋體">波長，用空白孔調零之后測各孔</font><font face="Calibri">OD</font><font face="宋體">值。</font></span><span style="mso-spacerun:'yes';font-family:宋體;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>