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<div id="gk2c0sac" class="appMask"></div> <div id="gk2c0sac" class="appContent"> <div id="gk2c0sac" class="phx-panel-wrap" data-showtitle="yes" data-isad="no" data-pattern="fixed" data-place="6" data-width="200" data-height="200"> <div id="gk2c0sac" class="phx-panel-title clearfix"> <span id="gk2c0sac" class="text pull-left">文章詳情</span> </div> <div id="gk2c0sac" class="phx-panel"> <div id="gk2c0sac" class="appWrapper" id="app_24330443" role="draggable" data-type="app" data-appid="datumInfo" data-config="" data-state="true"> <div id="gk2c0sac" class="appMask"></div> <div id="gk2c0sac" class="appContent"> <div id="gk2c0sac" class="datum_info"> <h1 class="title">ELISPOT技術(shù)的優(yōu)勢</h1> <div id="gk2c0sac" class="bgcolor_01"> <div id="gk2c0sac" class="bgcolor_sum"> <div id="gk2c0sac" class="left">日期：2024-11-22 05:59</div> <div id="gk2c0sac" class="right">瀏覽次數(shù)：744</div> </div> <div id="gk2c0sac" class="bgcolor_sum01"> 摘要：酶聯(lián)**斑點(diǎn)（ELISPOT）技術(shù)是當(dāng)今世界上檢測生物體細(xì)胞**水平的*佳技術(shù)。它集高靈敏度、高可信度、高通量、單細(xì)胞水平、功能性檢測與低成本等諸多優(yōu)點(diǎn)于一身，在國內(nèi)外**學(xué)界獲得了廣泛的應(yīng)用，成為主流**學(xué)檢測技術(shù)之一。我們來一一詳述這些優(yōu)點(diǎn)。靈敏度高理論上，ELISPOT高達(dá)百萬分之一，也就是說哪怕一百萬個(gè)陰性細(xì)胞中只有一個(gè)陽性細(xì)胞，ELISPOT能把這個(gè)陽性細(xì)胞檢測出來。說來也沒什么神奇之處，只要細(xì)胞狀態(tài)好，陰性細(xì)胞就不會產(chǎn)生斑點(diǎn)。而有一個(gè)陽性細(xì)胞，就會產(chǎn)生一個(gè)斑點(diǎn)，反過來，一個(gè)斑點(diǎn)就證明了一個(gè)... </div> </div> <div id="gk2c0sac" class="content"> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> 酶聯(lián)**斑點(diǎn)（ELISPOT）技術(shù)是當(dāng)今世界上檢測生物體細(xì)胞**水平的*佳技術(shù)。它集高靈敏度、高可信度、高通量、單細(xì)胞水平、功能性檢測與低成本等諸多優(yōu)點(diǎn)于一身，在國內(nèi)外**學(xué)界獲得了廣泛的應(yīng)用，成為主流**學(xué)檢測技術(shù)之一。我們來一一詳述這些優(yōu)點(diǎn)。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">靈敏度高</strong> <br> 理論上，ELISPOT高達(dá)百萬分之一，也就是說哪怕一百萬個(gè)陰性細(xì)胞中只有一個(gè)陽性細(xì)胞，ELISPOT能把這個(gè)陽性細(xì)胞檢測出來。說來也沒什么神奇之處，只要細(xì)胞狀態(tài)好，陰性細(xì)胞就不會產(chǎn)生斑點(diǎn)。而有一個(gè)陽性細(xì)胞，就會產(chǎn)生一個(gè)斑點(diǎn)，反過來，一個(gè)斑點(diǎn)就證明了一個(gè)陽性細(xì)胞的存在，樣哪怕這個(gè)陽性細(xì)胞混雜在一百萬個(gè)陰性細(xì)胞中，ELISPOT能把這個(gè)陽性細(xì)胞檢測出來。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> 實(shí)踐中，由于一個(gè)ELISPOT孔容納不了100萬細(xì)胞，*多只能放<u style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">50-60</u>萬細(xì)胞。再者，50-60萬陰性細(xì)胞出一兩個(gè)自發(fā)斑點(diǎn)也算正常，所以實(shí)驗(yàn)組的孔要想有顯著的統(tǒng)計(jì)學(xué)意義，需要出4-5個(gè)斑點(diǎn)。這也就是說，只要陽性細(xì)胞頻率大于<strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">4個(gè)/60萬細(xì)胞</strong>，就可以被ELISPOT檢測出來。這算出來的實(shí)際靈敏度為<strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">十五萬分之一。</strong>這樣的靈敏度在一般客戶的實(shí)驗(yàn)室都能達(dá)到，如果實(shí)驗(yàn)再精細(xì)一些，細(xì)胞處理得再好一些，靈敏度還可以提高一倍，達(dá)到<strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">三十萬分之一</strong>。作為對比，采用流式細(xì)胞儀的方法（細(xì)胞內(nèi)細(xì)胞因子染色法），對于陽性細(xì)胞的檢測靈敏度只有千分之一，比ELISPOT技術(shù)要低兩個(gè)數(shù)量級！ </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">單細(xì)胞水平</strong> <br> 與流式技術(shù)一樣，ELISPOT也是一種單細(xì)胞水平的檢測。作為對比，ELISA法、51Cr釋放法、細(xì)胞增殖實(shí)驗(yàn)法都是群體水平的檢測。如果群體細(xì)胞因子水平提高，既有可能是分泌細(xì)胞因子的細(xì)胞數(shù)量增加，也有可能是分泌細(xì)胞的數(shù)量沒增加而單個(gè)分泌水平增加。這種區(qū)別對研究生物體的細(xì)胞**是很重要的，但是群體水平的檢測不能區(qū)別出來，只有單細(xì)胞水平的檢測能夠勝任。也就是說，單細(xì)胞水平的檢測可以提供更多的信息。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">功能性檢測</strong> <br> ELISPOT檢測到的細(xì)胞因子不是實(shí)現(xiàn)存在的，而是在檢測的過程中，被加入的刺激物刺激而分泌出來的。斑點(diǎn)的強(qiáng)弱和數(shù)量反映了細(xì)胞對刺激物的反應(yīng)能力和分泌細(xì)胞因子的能力，這是體現(xiàn)細(xì)胞**反應(yīng)的*重要的功能指標(biāo)。作為對比，ELISA只能檢測細(xì)胞的**物，充其量就是間接的反映了細(xì)胞功能。而流式細(xì)胞內(nèi)細(xì)胞因子染色法需要的把本該分泌出來的細(xì)胞因子人為的囚禁在細(xì)胞之內(nèi)，因能反映細(xì)胞應(yīng)對刺激生產(chǎn)細(xì)胞因子的能力，不能反映其分泌細(xì)胞因子的能力，畢竟細(xì)胞因子要分泌到包外才能發(fā)揮功能活力。另外，流式方法也不能去除一類假陽性細(xì)胞，這類細(xì)胞的細(xì)胞因子含量豐富，但是沒有分泌功能。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">實(shí)現(xiàn)高通量檢測</strong> <br> ELISPOT是在96孔微孔板上進(jìn)行的，是一種高通量的檢測與篩選方法。一位有經(jīng)驗(yàn)的實(shí)驗(yàn)者一次可以處理上百份細(xì)胞樣品與數(shù)十種抗原，測定它們的反應(yīng)強(qiáng)度。作為對比，流式方法的樣品細(xì)胞需要在獨(dú)立的離心管中進(jìn)行，遠(yuǎn)未實(shí)現(xiàn)高通量，工作效率要低很多。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">直觀，可信度高</strong> <br> 相對ELISA法、流式細(xì)胞因子染色法、測CTL的51Cr釋放法而言，ELISPOT是*直觀的方法，一個(gè)陽性細(xì)胞一個(gè)斑點(diǎn)，清清楚楚，毫無疑問。只要實(shí)驗(yàn)遵循標(biāo)準(zhǔn)化的操作方法，該檢測的重復(fù)性很好。另外，ELISPOT檢測的樣品細(xì)胞可以在液氮里面長期冷凍存放，**（或者**）前后的細(xì)胞可以做對比檢測，細(xì)胞**水平的變化一目了然，可信度高。 </p> <p style="margin-top:0px;margin-bottom:1.4em;padding:0px;white-space:normal;border:0px;outline:0px;font-size:14px;vertical-align:baseline;line-height:25px;color:#333333;font-family:Arial, nsimsun, 'Microsoft YaHei', sans-serif;background-color:#FFFFFF;"> <strong style="margin:0px;padding:0px;border:0px;outline:0px;vertical-align:baseline;background-color:transparent;">檢測成本低</strong> <br> 流式方法需要昂貴復(fù)雜的流式細(xì)胞儀，但是ELISPOT檢測并不需要特殊的儀器設(shè)備，充其量在檢測規(guī)模擴(kuò)大了之后需要購置全自動的斑點(diǎn)圖像分析儀。另外，ELISPOT試劑相對于ELISA與流式抗體也要便宜很多。達(dá)科為公司代理的進(jìn)口U-CyTech未包被ELISPOT試劑盒，96test價(jià)格在1000元左右，達(dá)科為公司推出的Quick Spot系列預(yù)包被ELISPOT試劑盒的價(jià)格也不到2000元。可以說，ELISPOT是一項(xiàng)功能強(qiáng)大、物美價(jià)廉的檢測技術(shù)。 </p> </div> <div id="gk2c0sac" class="datumrelate"> <span>下一篇：</span> <a href="/klbscience_Article_7424711.html" title="ELISPOT檢測的質(zhì)量控制">ELISPOT檢測的質(zhì)量控制</a> <br> <span>上一篇：</span> <a href="/klbscience_Article_7576549.html" 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