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<div class="qo0ssqw6" id="layout_20040304_style" class="hide"> </div> <div id="qo0ssqw6" class="appMask"></div> <div id="qo0ssqw6" class="appContent"> <div id="qo0ssqw6" class="phx-panel-wrap" data-showtitle="yes" data-isad="no" data-pattern="fixed" data-place="6" data-width="200" data-height="200"> <div id="qo0ssqw6" class="phx-panel-title clearfix"> <span id="qo0ssqw6" class="text pull-left">文章詳情</span> </div> <div id="qo0ssqw6" class="phx-panel"> <div id="qo0ssqw6" class="appWrapper" id="app_24330443" role="draggable" data-type="app" data-appid="datumInfo" data-config="" data-state="true"> <div id="qo0ssqw6" class="appMask"></div> <div id="qo0ssqw6" class="appContent"> <div id="qo0ssqw6" class="datum_info"> <h1 class="title">血清對(duì)ELISPOT檢測(cè)的干擾</h1> <div id="qo0ssqw6" class="bgcolor_01"> <div id="qo0ssqw6" class="bgcolor_sum"> <div id="qo0ssqw6" class="left">日期：2024-11-22 05:59</div> <div id="qo0ssqw6" class="right">瀏覽次數(shù)：582</div> </div> <div id="qo0ssqw6" class="bgcolor_sum01"> 摘要：一直以來，ELISPOT檢測(cè)對(duì)于血清的影響都是眾說紛紜，利弊各執(zhí)一詞。一方面，**細(xì)胞在體外的存活離不開血清的營養(yǎng)， ELISPOT檢測(cè)中細(xì)胞要在孔中孵育一兩天，所以培養(yǎng)基中需要添加血清；另一方面，血清中含有許許多多未知成分能夠抑制或者額外刺激**細(xì)胞從而干擾ELISPOT檢測(cè)，尤其在針對(duì)一些臨床樣本的ELISPOT檢測(cè)中影響更不容忽視！由于血清是半天然的生物制品，它的質(zhì)量受采集的動(dòng)物個(gè)體差異的影響。所以，即使同一個(gè)公司同**別的血清產(chǎn)品，不同批次之間對(duì)ELISPOT的影響也是各不相同的。血清這種或者抑制或者額外刺激斑點(diǎn)... </div> </div> <div id="qo0ssqw6" class="content"> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;text-indent:24pt;background-color:#FFFFFF;"> <span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">一直以來，ELISPOT檢測(cè)對(duì)于血清的影響都是眾說紛紜，利弊各執(zhí)一詞。一方面，**細(xì)胞在體外的存活離不開血清的營養(yǎng)， ELISPOT檢測(cè)中細(xì)胞要在孔中孵育一兩天，所以培養(yǎng)基中需要添加血清；另一方面，血清中含有許許多多未知成分能夠抑制或者額外刺激**細(xì)胞從而干擾ELISPOT檢測(cè)，尤其在針對(duì)一些臨床樣本的ELISPOT檢測(cè)中影響更不容忽視！由于血清是半天然的生物制品，它的質(zhì)量受采集的動(dòng)物個(gè)體差異的影響。所以，即使同一個(gè)公司同**別的血清產(chǎn)品，不同批次之間對(duì)ELISPOT的影響也是各不相同的。</span><span style="font-size:12pt;font-family:微軟雅黑;"></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;text-indent:24pt;background-color:#FFFFFF;"> <span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">血清這種或者抑制或者額外刺激斑點(diǎn)的性質(zhì)，很讓人困惑。以前人們推測(cè)是因?yàn)檠謇锩婧懈鞣N牛的細(xì)胞因子，它與其它種屬的細(xì)胞因子受體有交叉反應(yīng)，可以激活或者抑制相應(yīng)的待檢測(cè)細(xì)胞?，F(xiàn)在對(duì)于TLR受體及其信號(hào)機(jī)制了解更深刻之后，似乎內(nèi)源性TLR供體是一個(gè)更可能的原因。TLR供體不僅僅指來源于外源的病原體，哺乳動(dòng)物自身可以表達(dá)一系列的TLR供體而激活TLR信號(hào)通路。這些內(nèi)源性的TLR供體</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">包括IL-1α、IL-1β、β-defensin2、HSP60、HSP70、Fibirinogen、透明質(zhì)酸上的寡糖（Oligosaccharides of hyal</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">ronic acid）、一種纖維連接蛋白上的結(jié)構(gòu)域（Type III repeat extra domain A of fibronectin）、肝素上的多糖（Polysaccharide fragment of heparan s</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">lphate）、CpG染色質(zhì)IgG2a復(fù)合體。</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">（Jiang, Z. et. al., 2003；B</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">l</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">t, Y. et. al., 2001；Biragyn, A. et. al., 2002；Okam</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">ra, Y. et. al., 2001；Johnson, G. B. et. al., 2002；X</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">u</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">, H. et. al., 2003）</span><span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">。</span><span style="font-size:12pt;font-family:微軟雅黑;"></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;text-indent:24pt;background-color:#FFFFFF;"> <span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">為什么說，內(nèi)源性TLR供體比細(xì)胞因子更可能對(duì)胎牛血清的干擾效應(yīng)負(fù)責(zé)呢？因?yàn)槲覀冎兰?xì)胞因子及其受體的種屬特異性是很強(qiáng)的，并且細(xì)胞因子的穩(wěn)定性差，容易失活。而相應(yīng)的內(nèi)源性TLR供體——比如HSP60/70、透明質(zhì)酸上的寡糖、肝素上的多糖——在進(jìn)化上要保守得多，種屬間沒有什么差異。所以牛表達(dá)的這些TLR供體刺激人或者鼠或者猴子的白細(xì)胞上的TLR幾乎與種屬內(nèi)的刺激有相同的效果。另外，牛TLR供體的穩(wěn)定性好得多，不會(huì)在血清保存或者滅活補(bǔ)體的過程中失活。</span><span style="font-size:12pt;font-family:微軟雅黑;"></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;text-indent:24pt;background-color:#FFFFFF;"> <span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">在這近20年的時(shí)間里，人們只能采用*繁瑣的辦法來努力消除血清的干擾。具體做法是從全世界不同的血清供應(yīng)商那里索取各種不同批次的血清樣品，然后逐一做ELISPOT對(duì)比實(shí)驗(yàn)，找出干擾*小、重復(fù)性*好的血清品種與批次。然后就大量購買該品種、該批次的血清，以后每次都用它做檢測(cè)。直至該批次血清用盡，然后再開始下一輪的比較與采購。為了ELISPOT檢測(cè)結(jié)果的連貫性，通常會(huì)保留少許上批次的血清，用以作下輪比較的參照。</span><span style="font-size:12pt;font-family:微軟雅黑;"></span> </p> <p style="margin-top:0px;margin-bottom:0px;padding:0px;font-family:'Microsoft Yahei', arial, 宋體;line-height:21.6px;white-space:normal;text-indent:24pt;background-color:#FFFFFF;"> <span style="font-size:12pt;font-family:微軟雅黑;color:#333333;">血清就是這樣困擾著ELISPOT技術(shù)將近20年。人們一直渴望有一種化學(xué)成分完全限定，對(duì)ELISPOT沒有任何干擾的，批次間影響完全相同的培養(yǎng)基，以徹底消除血清對(duì)ELISPOT技術(shù)的干擾，于是就催生了無血清ELISPOT技術(shù)。在*近兩三年的進(jìn)步與實(shí)踐中，無血清ELISPOT技術(shù)經(jīng)歷諸多考驗(yàn)，已經(jīng)成熟。</span> </p> </div> <div id="qo0ssqw6" class="datumrelate"> <span>下一篇：</span> <a href="/klbscience_Article_6947248.html" title="ELISPOT斑點(diǎn)的特征">ELISPOT斑點(diǎn)的特征</a> <br> <span>上一篇：</span> <a href="/klbscience_Article_7078264.html" title="無血清ELISPOT技術(shù)">無血清ELISPOT技術(shù)</a> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <footer id="footer" class="footer" data-appid="layout"> <div class="qo0ssqw6" id="footerWrapper" class="footerWrapper" data-appid="wrapper"> <div class="qo0ssqw6" id="footerContainer" class="footerContainer" data-appid="container"> <div class="qo0ssqw6" id="app_55124555_style" class="hide"> </div> <div id="qo0ssqw6" class="row" role="draggable" data-appid="row" id="layout_50125041"> <div class="qo0ssqw6" id="layout_50125041_style" class="hide"> </div> <div id="qo0ssqw6" class="rowPatch"> <div id="qo0ssqw6" class="col-12"> <div id="qo0ssqw6" class="boxBlock"> <div id="qo0ssqw6" class="elementContainer" data-appid="column" id="layout_01500040"> <div id="qo0ssqw6" class="appWrapper" id="app_55124555" role="draggable" data-type="app" data-appid="footer" data-config="" data-state="true"> <div id="qo0ssqw6" class="appMask"></div> <div id="qo0ssqw6" class="appContent"> <div id="qo0ssqw6" class="phx-footer"> <p> Copyright? 2024 上?？道噬锟萍加邢薰?/p> <p> 電話：021-61998208 傳真：021-61998551 地址：郵編：</p> <p> <a href="/promotion/default.htm" target="_blank" rel="nofollow" class="buttoma">易展提供技術(shù)支持</a> <a target="_blank" rel="nofollow">滬ICP備15039594號(hào)-2</a> <a target="_blank" rel="nofollow">網(wǎng)站管理入口</a> <a rel="nofollow" target="_blank" class="yz-trustlogo"></a> </p> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </footer> <div style="width:300px;margin:0 auto; 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