ELISA 检测的目的是为实验提供准确可靠的定量分析实验依据。为了保证实验数据的可靠性，在实验过程中必须坚持全面质量控制和全过程质量控制。在收集标本前都必须有一个完整的计划。齐一生物销售：021－6034 8496；181214 53965；173021 04490

注意事项：

◆   每个标本量收集体积＝  100ul × 检测种类，如要做复孔，标本量收集体积＝  100ul × 检测种类×2 。

◆   对于作某一个具体指标来说，先用一系列稀释度作一批标本测定，得出对实验有意义的一个稀释度（即测定剂量反应曲线），然后用一个适稀释度测定即可。

◆   收集标本前必须清楚要检测的成份是否足够稳定。对收集后当天进行检测的标本，储存在4℃备用，如有特殊原因需要周期收集标本，将标本及时分装后放在-20℃ 或-70℃ 条件下保存。避免反复冻融。标本 2 -8 ℃ 可保存48 小时.-20℃ 可保存1个月。 -70 度可保存6个月。部分激素类标本需添加抑肽酶。

◆   血清标本采集时，应注意避免溶血，红细胞溶解时会释放出具有过氧化物酶活性的物质，以 HRP为标记的 ELISA 测定中，溶血标本可能会增加非特异性显色。

【人流感病毒B（FLUB）ELISA试剂盒】◆   为了保证尿液检测结果的准确性，必须正确收集尿液标本和保存。盛尿容器要清洁干燥。使用一次性的容器（如塑料尿杯），避免因用药并清洗不干净而造成的污染，影响检测结果。尿液标本必须新鲜，留取后，应及时检测或保存，以免细菌繁殖。因在室温中久置后（尤其是夏季）尿中磷酸盐等可析出结晶而干扰检测。

◆    标本宜在新鲜时检测。如有细菌污染，菌体中可能含有内源性 HRP，也会产生假阳性反应。保存过久可发生聚合在 ELISA中可使本底加深。

◆   冻结标本融解后，蛋白质局部浓缩，分布不均，应充分混匀宜轻缓，避免气泡，可上下颠倒混和，不要在混匀器上强烈振荡。

◆   混浊或有沉淀的标本应先离心或过滤，澄清后再检测。

◆   反复冻融会使蛋白效价降低，所以待测标本如需保存作多次检测，宜少量分装冰存。也可加入适当防腐剂。

◆   如果您在实验中遇到任何特殊问题，敬请咨询 021-60348496标本类型

ELISA常见的标本一般包括：

◆   液体类标本：包括血清、血浆、尿液、胸腹水、脑脊液、细胞培养上清等。

◆   培养细胞

◆   组织标本

这些标本收集的时间、方法和保存都有一定的要求。

【人流感病毒B（FLUB）ELISA试剂盒】标本的保存

一般说来，在 5 天内测定的标本可放置于 4℃，超过一周测定的需低温冻存。

对收集后当天进行检测的标本，储存在  4 ℃ 备用，如有特殊原因需要周期收集标本， 将标本及时分装后放在  -20 ℃ 或  -70 ℃ 条件下保存。避免反复冻融。 标本  2 -8 ℃ 可保存  48 小时，-20 ℃ 可保存  1 个月。-70 度可保存  6 个月。部分标本也可加入适当防腐剂，激素类标本需添加抑肽酶。标本的处理 可用作 ELISA 测定的标本十分广泛，体液（如血清）、分泌物（唾液）和排泄物（如尿液）、细胞培养上清、细胞、组织等均可作标本以测定其中某种成份。有些标本可直接进行测定，有些则需经预处理。

◆ 血清：  

室温血液自然凝固  10-20 分钟后，离心  20 分钟左右（  2000-3000 转  / 分）。收集上清。如有沉淀形成，应再次离心。  

◆ 血浆：  

应根据试剂盒的要求选择  EDTA 、柠檬酸钠或肝素作为抗凝剂，加入  10 ％（  v/v ）抗凝剂 ( 0.1M 柠檬酸钠或  1% heparin 或  2.0%EDTA.Na2) 混合  10-20 分钟后，离心  20 分钟左右（  2000-3000 转  / 分）。仔细收集上清如有沉淀形 成，应再次离心。  

【人流感病毒B（FLUB）ELISA试剂盒】◆ 尿液、胸腹水、脑脊液：  

用无菌管收集。离心  20 分钟左右（  2000-3000 转  / 分）。仔细收集上清。如有沉淀形成，应再次离心。  

◆ 细胞培养上清：  

检测分泌性的成份时，用无菌管收集。离心  20 分钟左右（  2000-3000 转  / 分）。仔细收集上清。检测细胞内的 成份时，用  PBS （  PH7.2-7.4 ）稀释细胞悬液，细胞浓度达到  100 万 /ml 左右。通过反复冻融，以使细胞破坏并放出细胞内成份。离心  20 分钟左右（  2000-3000 转 / 分）。仔细收集上清。保存过程中如有沉淀形成，应再次离心。  

◆ 细胞：

检测细胞的成份时，对于贴壁细胞，去除培养液，用PBS、生理盐水或无血清培养液洗一遍。加入适量裂解液。用枪吹打数下，使裂解液和细胞充分接触。通常裂解液接触细胞10 秒后，细胞就会被裂解。对于悬浮细胞，离心收集细胞，用PBS、生理盐水或无血清培养液洗一遍.加入适量裂解液, 用枪吹打把细胞吹散。用手指轻弹以充分裂解细胞。充分裂解后应没有明显的细胞沉淀。如果细胞量较多，必需分装然后再裂解。充分裂解后，10000-14000g离心3-5 分钟，取上

清，即可进行后续操作。

【人流感病毒B（FLUB）ELISA试剂盒】◆ 组织标本：  

切割标本后，称取重量。加入一定量的  PBS或组织蛋白萃取试剂，缓冲液中可加入蛋白酶抑制剂。用手工或匀浆器将标本匀浆充分。离心  20 分钟左右（  2000-3000 转  / 分）。仔细收集上清置于  -20 度或  - 70 度保存，如有必要，可以将样品浓缩干燥。分装后一份待检测，其余冷冻备用。齐一生物销售：021－6034 8496；181214 53965；173021 04490  

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