如何選擇合適的 ELISA 試劑盒
<p> <span style="font-family: 宋體; font-size: 10.5pt;"> </span><font face="宋體" style="font-family: 宋體; font-size: 10.5pt;">酶聯(lián)免疫吸附試驗(</font><font face="Calibri" style="font-size: 10.5pt;">ELISA</font><font face="宋體" style="font-family: 宋體; font-size: 10.5pt;">)是指將可溶性抗原或抗體吸附于聚苯乙烯等固相載體上,進行免疫反應,以測定液體生物標本中微量物質的一種定性、定量方法。</font></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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="Calibri">ELISA</font><font face="宋體">檢測具有靈敏度高、特異性強、操作簡單、樣本量小等特點,且檢測結果準確性高、重復性好,廣泛應用于臨床和科研中,用于抗原抗體檢測以及病原體檢測。目前,市面上</font><font face="Calibri">ELISA</font><font face="宋體">試劑盒品牌眾多,檢測指標種類繁多,產品別名繁多,產品質量良莠不齊。我們應該如何選擇一款合適的</font><font face="Calibri">ELISA</font><font face="宋體">試劑盒產品?需要考慮哪些指標?</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">首先,用戶需要明確實驗的目的是功能測定還是目標分子的含量檢測。功能測定一般是酶與底物的反應,分析生物活性。</font><font face="Calibri">Elisa</font><font face="宋體">則專門用于目標分子的含量檢測。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">其次,如果研究的索引是蛋白質或者多肽,可以到</font><font face="Calibri">Uniprot</font><font face="宋體">數(shù)據(jù)庫查詢對應的</font><font face="Calibri">UniprotID</font><font face="宋體">,選擇產品時同時查看索引名稱和</font><font face="Calibri">UniprotID</font><font face="宋體">;如果研究的索引是小分子半抗原,這類化學物質可以到</font><font face="Calibri">ChemSpider</font><font face="宋體">化學品庫中查詢對應的</font><font face="Calibri">CAS</font><font face="宋體">號,根據(jù)</font><font face="Calibri">CAS</font><font face="宋體">號選擇需要的產品。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">最后,我們還需要根據(jù)現(xiàn)有的文獻查詢或預測靶標在生物樣本中的存在情況或含量范圍,選擇具有足夠靈敏度的試劑盒,并與廠家確認該試劑盒的適用樣本是否包括客戶自身的實驗樣本。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">在此基礎上,如果研究人員有多個品牌的同類產品可供選擇,還應該比較試劑盒產品的多項性能參數(shù),包括靈敏度、特異性、精密度、準確度、線性、穩(wěn)定性以及自然樣本數(shù)據(jù)。這些參數(shù)是</font><font face="Calibri">ELISA</font><font face="宋體">作為一種人工建立的微量分析方法必須考慮的固有特性,以確保其有效性和可靠性。</font></span></p>
<p class="MsoNormal"> </p>
<p class="MsoNormal" style="text-align: center;"><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:10.5000pt; mso-font-kerning:1.0000pt;"><font face="宋體"><img src="/images/upload/Image/微信圖片_20250331165121.jpg" alt="elisa試劑盒" width="500" height="375" /></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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">敏感度</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">靈敏度</font><font face="宋體">——試劑盒對待測物質的最低檢測能力。如果待測物質的濃度很低,我們可以選擇靈敏度較高的試劑盒。靈敏度值是空白對照標準差對應濃度值的</font><font face="Calibri">3</font><font face="宋體">倍。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">特異性</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">特異性決定了其他分子與試劑盒抗體結合的能力。交叉反應性越小,試劑盒的特異性越好。通常需要測定試劑盒是否與當前物種同家族的蛋白、其他物種的蛋白、血清中的常見因子(如嗜異性抗體、類風濕因子等)以及靶蛋白相互作用蛋白無交叉反應。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">精確</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">精密度</font><font face="宋體">——又稱重復性,是對同一樣品進行重復測量時誤差大小的評價參數(shù)。一般要求板間、板內精密度控制在</font><font face="Calibri">10%</font><font face="宋體">以內。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">恢復</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">回收率</font><font face="宋體">——通常以加標回收率來衡量,回收率在</font><font face="Calibri">80%</font><font face="宋體">到</font><font face="Calibri">120%</font><font face="宋體">之間為合格。測量方法是將標準品添加到天然樣品基質中。未添加標準品的天然樣品基質作為空白對照?;厥章手凳悄繕宋锏臏y得濃度與添加標準品濃度的比值,以百分比表示?;厥章蔬^高或過低都會產生假陽性或假陰性結果,影響測量值的準確性。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">線性</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">將適當濃度的標準品加入天然樣品基質中,稀釋</font><font face="Calibri">2</font><font face="宋體">倍、</font><font face="Calibri">4</font><font face="宋體">倍、</font><font face="Calibri">8</font><font face="宋體">倍進行檢測,測定各濃度下的回收率,回收率在</font><font face="Calibri">80%~120%</font><font face="宋體">之間為合格。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">穩(wěn)定</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">驗證一般采用加速破壞性實驗,將未開封的試劑盒分別在</font><font face="Calibri">37</font><font face="宋體">℃和</font><font face="Calibri">2-8</font><font face="宋體">℃條件下放置一定時間,考察各項性能指標的偏差,偏差在</font><font face="Calibri">10%</font><font face="宋體">以內為合格。穩(wěn)定性越好,試劑盒可以保存的時間越長。</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">天然樣品檢測</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:10.5000pt; 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:10.5000pt; mso-font-kerning:1.0000pt;"> </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:10.5000pt; mso-font-kerning:1.0000pt;"> <font face="宋體">最后,還應明確試劑盒可檢測的天然樣本及適用的樣本類型。根據(jù)目標的性質預測適合檢測的樣本,并實際檢測</font><font face="Calibri">5</font><font face="宋體">個以上該類型的樣本,并在客戶需要時提供一定的樣本含量數(shù)據(jù)參考。</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:10.5000pt; mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>