ELISA檢測方法：四種ELISA類型優(yōu)缺點比較

<p>&nbsp;<font face="宋體" style="font-size: 10.5pt;">　　酶聯(lián)免疫吸附實驗（</font><span style="font-family: Calibri; font-size: 10.5pt;">ELISA</span><font face="宋體" style="font-size: 10.5pt;">）是一種成熟且廣泛應用的靶標檢測方法，以其操作簡便、安全和高特異性而著稱。通過抗原抗體特異性結(jié)合和酶促反應，</font><font face="Calibri" style="font-family: Calibri; font-size: 10.5pt;">ELISA</font><font face="宋體" style="font-size: 10.5pt;">可實現(xiàn)定性和定量檢測。根據(jù)原理，</font><font face="Calibri" style="font-family: Calibri; font-size: 10.5pt;">ELISA</font><font face="宋體" style="font-size: 10.5pt;">可分為直接、間接、夾心和競爭四種方法。本文將深入探討這四種</font><font face="Calibri" style="font-family: Calibri; font-size: 10.5pt;">ELISA</font><font face="宋體" style="font-size: 10.5pt;">類型的特點及應用。</font></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <h3><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　</font>ELISA<font face="宋體">檢測方法</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></h3> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　</font>ELISA<font face="宋體">（酶聯(lián)免疫吸附測定）檢測方法主要包括以下</font><font face="Calibri">4</font><font face="宋體">種類型，每種方法適用于不同的檢測需求和目標分子特性：</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　1、直接法</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　抗原直接固定在固相載體上，然后加入特異性酶標抗體進行結(jié)合。不需二抗，直接通過酶標抗體與抗原的結(jié)合進行檢測。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　優(yōu)點：步驟簡單，速度快，減少交叉反應。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　缺點：背景可能較高，每種抗原需要專門的酶標抗體，靈敏度較低。</font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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" style="text-align: left;"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;<img src="/images/upload/Image/直接ELISA(1).png" alt="ELISA" width="216" height="111" /></span></p> <p class="MsoNormal" style="text-align: left;">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　2、間接法</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　將未標記的特異性抗體與固相載體上的抗原結(jié)合，之后加入酶標二抗（針對抗體的抗體）進行檢測。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　優(yōu)點：靈敏度高，經(jīng)濟，靈活性高，因為一個酶標二抗可以用于多種一抗。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　缺點：可能有交叉反應，實驗周期較長。</font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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/間接ELISA.png" alt="ELISA" width="216" height="170" /></font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　3、雙抗夾心法</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　首先用捕獲抗體包被固相載體，加入樣本后，抗原被捕捉，再加入酶標檢測抗體，形成抗體</font>-<font face="宋體">抗原</font><font face="Calibri">-</font><font face="宋體">酶標抗體復合物。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　優(yōu)點：特異性和敏感性高，適合檢測大分子抗原。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　缺點：需要兩種特異性抗體，制備成本相對高。</font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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/夾心ELISA(1).png" alt="ELISA" width="216" height="197" /></font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　4、競爭法</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　抗原預先包被在固相載體上，樣本中的抗原與酶標抗原競爭結(jié)合固相抗原，然后通過底物顯色來間接反映樣本中抗原的量。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　優(yōu)點：適用于小分子抗原的檢測，能處理較復雜或不純的樣本。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　缺點：操作較復雜，敏感性和特異性可能因競爭而降低。</font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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/競爭ELISA(1).png" alt="ELISA" width="216" height="159" /></font></span></p> <p class="MsoNormal">&nbsp;</p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　上述是</font>4<font face="宋體">種</font><font face="Calibri">ELISA</font><font face="宋體">類型，每種檢測方法都有各自優(yōu)缺點，每種方法的選擇取決于待測物質(zhì)的性質(zhì)、實驗目的以及實驗室的具體條件。直接法和競爭法因其特定的局限性應用較少，而間接法和雙抗夾心法由于其靈活性和高靈敏度，在實際應用中更為常見。隨著技術(shù)的不斷發(fā)展，</font><font face="Calibri">ELISA</font><font face="宋體">技術(shù)也在不斷改進，例如新型標記物和檢測方法的出現(xiàn)，使其在生物醫(yī)學研究和臨床診斷中發(fā)揮著越來越重要的作用。</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;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:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;">&nbsp;</span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><font face="宋體">　　</font></span><span style="mso-spacerun:'yes';font-family:Calibri;mso-fareast-font-family:宋體;&#10;mso-bidi-font-family:'Times New Roman';font-size:10.5000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>