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技術(shù)文章您的位置:網(wǎng)站首頁 >技術(shù)文章 > 注意??!樣本保存不當(dāng),也很容易影響ELISA實(shí)驗(yàn)結(jié)果!
注意??!樣本保存不當(dāng),也很容易影響ELISA實(shí)驗(yàn)結(jié)果!
更新時(shí)間:2016-08-17   點(diǎn)擊次數(shù):1625次

注意!!樣本保存不當(dāng),也很容易影響ELISA實(shí)驗(yàn)結(jié)果!

大家在做實(shí)驗(yàn)的時(shí)候,樣本的收集和保存尤為重要,樣本收集保存不當(dāng),可能對(duì)您的結(jié)果產(chǎn)生很嚴(yán)重的影響,下面讓我們信帆生物的技術(shù)給大家講解一下樣本應(yīng)該怎么保存和收集!

(1)標(biāo)本溶血 由于各種人為原因引起的標(biāo)本溶血,均可因紅細(xì)胞破壞溶解時(shí)釋放出大量具有過氧化物酶活性的血紅蛋白,在以辣根過氧化物酶為標(biāo)記的 ELISA 測(cè)定中,會(huì)導(dǎo)致非特異性顯色,干擾測(cè)定結(jié)果。為克服上述干擾作用,標(biāo)本采集時(shí)必須注意避免溶血。

(2)標(biāo)本受細(xì)菌污染 因菌體中可能含有內(nèi)源性辣根過氧化物酶,因此,被細(xì)菌污染的標(biāo)本同溶血標(biāo)本一樣,亦可產(chǎn)生非特異性顯色而干擾測(cè)定結(jié)果。

(3)標(biāo)本保存不當(dāng) 在冰箱中保存過久的標(biāo)本,血清中 IgG 可聚合成多聚體、AFP 可形成二聚體,在間接法 ELISA 測(cè)定中會(huì)導(dǎo)致本底過深、甚至造成假陽性;標(biāo)本放置時(shí)間過長(如一天以上),有時(shí)抗原或抗體免疫活性減弱,亦可出現(xiàn)假陰性。為克服上述干擾,ELISA 測(cè)定的血清標(biāo)本宜為新鮮采集;如不能立即測(cè)定,5 天內(nèi)測(cè)定的血 清標(biāo)本可存放于 4℃,1 周后測(cè)定的血清標(biāo)本應(yīng)低溫凍存;凍存后融解的標(biāo)本,蛋白質(zhì)局部濃縮,分布不均,應(yīng)充分混合后再測(cè)定,但混勻時(shí)應(yīng)輕柔,不可強(qiáng)烈振 蕩。

(4)標(biāo)本凝集不全 在沒有促凝劑和抗凝劑存在的情況下,正常血液采集后 1/2~2 h 開始凝固,18~24 h *凝固。檢驗(yàn)工作中,有時(shí)為了爭取時(shí)間快速檢測(cè),常在血液還 未開始凝固時(shí)即強(qiáng)行離心分離血清,此時(shí)的血清中仍殘留部分纖維蛋白原,在 ELISA 測(cè)定過程中可以形成肉眼可見的纖維蛋白塊,易造成假陽性結(jié)果;這類情況 于次日復(fù)查時(shí)因血凝已*,血清中不再有纖維蛋白原存在,故復(fù)查結(jié)果變?yōu)殛幮?。為避免上述干擾作用,解決的辦法是血液標(biāo)本采集后必須使其充分凝固后再 分離血清,或標(biāo)本采集時(shí)用帶分離膠的采血管或于采血管中加入適當(dāng)?shù)拇倌齽?/p>

信帆生物代理銷售各類elisa試劑盒,提供各種實(shí)驗(yàn)代測(cè)服務(wù),歡迎大家!

 

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