實(shí)驗(yàn)時(shí)elisa競(jìng)爭(zhēng)法檢測(cè)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(shí)驗(yàn)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(shí)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(jìng)爭(zhēng)結(jié)合</font><font face="Calibri">HBeAg</font><font face="宋體">，被檢標(biāo)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(jié)合越多，而與包被抗</font><font face="Calibri">-HBe</font><font face="宋體">結(jié)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(nèi)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(yáng)性和陰性對(duì)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(jié)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(yáng)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(duì)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(xiàng)</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(shí)應(yīng)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(shí)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(dāng)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(yīng)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(hào)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(duì)照血清和陽(yáng)性對(duì)照血清于相應(yīng)孔中。設(shè)空白對(duì)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(nèi)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(fù)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(biāo)儀檢測(cè)：選用</font><font face="Calibri">450nm</font><font face="宋體">波長(zhǎng)，用空白孔調(diào)零之后測(cè)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>