如何选择合适的 ELISA 试剂盒

<p>&nbsp;<span style="font-family: 宋体; font-size: 10.5pt;">&nbsp;&nbsp;&nbsp;&nbsp;</span><font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">酶联免疫吸附试验（</font><font face="Calibri" style="font-size: 10.5pt;">ELISA</font><font face="宋体" style="font-family: 宋体; font-size: 10.5pt;">）是指将可溶性抗原或抗体吸附于聚苯乙烯等固相载体上，进行免疫反应，以测定液体生物标本中微量物质的一种定性、定量方法。</font></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="Calibri">ELISA</font><font face="宋体">检测具有灵敏度高、特异性强、操作简单、样本量小等特点，且检测结果准确性高、重复性好，广泛应用于临床和科研中，用于抗原抗体检测以及病原体检测。目前，市面上</font><font face="Calibri">ELISA</font><font face="宋体">试剂盒品牌众多，检测指标种类繁多，产品别名繁多，产品质量良莠不齐。我们应该如何选择一款合适的</font><font face="Calibri">ELISA</font><font face="宋体">试剂盒产品？需要考虑哪些指标？</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">首先，用户需要明确实验的目的是功能测定还是目标分子的含量检测。功能测定一般是酶与底物的反应，分析生物活性。</font><font face="Calibri">Elisa</font><font face="宋体">则专门用于目标分子的含量检测。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">其次，如果研究的索引是蛋白质或者多肽，可以到</font><font face="Calibri">Uniprot</font><font face="宋体">数据库查询对应的</font><font face="Calibri">UniprotID</font><font face="宋体">，选择产品时同时查看索引名称和</font><font face="Calibri">UniprotID</font><font face="宋体">；如果研究的索引是小分子半抗原，这类化学物质可以到</font><font face="Calibri">ChemSpider</font><font face="宋体">化学品库中查询对应的</font><font face="Calibri">CAS</font><font face="宋体">号，根据</font><font face="Calibri">CAS</font><font face="宋体">号选择需要的产品。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">最后，我们还需要根据现有的文献查询或预测靶标在生物样本中的存在情况或含量范围，选择具有足够灵敏度的试剂盒，并与厂家确认该试剂盒的适用样本是否包括客户自身的实验样本。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">在此基础上，如果研究人员有多个品牌的同类产品可供选择，还应该比较试剂盒产品的多项性能参数，包括灵敏度、特异性、精密度、准确度、线性、稳定性以及自然样本数据。这些参数是</font><font face="Calibri">ELISA</font><font face="宋体">作为一种人工建立的微量分析方法必须考虑的固有特性，以确保其有效性和可靠性。</font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal" style="text-align: center;"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><font face="宋体"><img src="/images/upload/Image/微信图片_20250331165121.jpg" alt="elisa试剂盒" width="500" height="375" /></font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">敏感度</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">灵敏度</font><font face="宋体">&mdash;&mdash;试剂盒对待测物质的最低检测能力。如果待测物质的浓度很低，我们可以选择灵敏度较高的试剂盒。灵敏度值是空白对照标准差对应浓度值的</font><font face="Calibri">3</font><font face="宋体">倍。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">特异性</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">特异性决定了其他分子与试剂盒抗体结合的能力。交叉反应性越小，试剂盒的特异性越好。通常需要测定试剂盒是否与当前物种同家族的蛋白、其他物种的蛋白、血清中的常见因子（如嗜异性抗体、类风湿因子等）以及靶蛋白相互作用蛋白无交叉反应。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">精确</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">精密度</font><font face="宋体">&mdash;&mdash;又称重复性，是对同一样品进行重复测量时误差大小的评价参数。一般要求板间、板内精密度控制在</font><font face="Calibri">10%</font><font face="宋体">以内。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">恢复</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">回收率</font><font face="宋体">&mdash;&mdash;通常以加标回收率来衡量，回收率在</font><font face="Calibri">80%</font><font face="宋体">到</font><font face="Calibri">120%</font><font face="宋体">之间为合格。测量方法是将标准品添加到天然样品基质中。未添加标准品的天然样品基质作为空白对照。回收率值是目标物的测得浓度与添加标准品浓度的比值，以百分比表示。回收率过高或过低都会产生假阳性或假阴性结果，影响测量值的准确性。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">线性</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">将适当浓度的标准品加入天然样品基质中，稀释</font><font face="Calibri">2</font><font face="宋体">倍、</font><font face="Calibri">4</font><font face="宋体">倍、</font><font face="Calibri">8</font><font face="宋体">倍进行检测，测定各浓度下的回收率，回收率在</font><font face="Calibri">80%~120%</font><font face="宋体">之间为合格。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">稳定</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">验证一般采用加速破坏性实验，将未开封的试剂盒分别在</font><font face="Calibri">37</font><font face="宋体">℃和</font><font face="Calibri">2-8</font><font face="宋体">℃条件下放置一定时间，考察各项性能指标的偏差，偏差在</font><font face="Calibri">10%</font><font face="宋体">以内为合格。稳定性越好，试剂盒可以保存的时间越长。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">天然样品检测</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;">&nbsp;&nbsp;&nbsp;&nbsp;<font face="宋体">最后，还应明确试剂盒可检测的天然样本及适用的样本类型。根据目标的性质预测适合检测的样本，并实际检测</font><font face="Calibri">5</font><font face="宋体">个以上该类型的样本，并在客户需要时提供一定的样本含量数据参考。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;&#10;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;&#10;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>