ELISA品牌细胞因子检测试剂盒质控指标的验证
摘要：由于ELISA实验的高度专一性，敏感性，以及极高的准确度，现已广泛应用于医疗卫生检测，疾病的临床诊断与检查，食品中有害物质残留的检定，辅助基因工程等方面。本课题通过ELISA双抗夹心法来测试研发的试剂盒对于肿瘤坏死因子-α;白介素-1β;白介素-6;血管细胞黏附分子-1的实验效果，并加以改进，使其完善。实验时主要使用控制变量法，来确认待改进因素。通过实验，我首先确认四种HRP效果的差异性在可控范围内。然后确认一抗包被的效果良好，而且二抗的效价没有降低。由之前的实验发现显色速度过快，并造成本底过高的问题。通过采用控制变量法，并设置HRP以及二抗稀释倍数的对照实验，来确定HRP以及二抗的最终稀释倍数来降低本底。最终达成完善细胞因子检测试剂盒目的。81043
毕业论文关键词：双抗夹心法；肿瘤坏死因子-α；白介素-1β；白介素-6；血管细胞黏附分子-1
Validation of quality control index of a brand cytokine test kit
Abstract：Because of the specificity of ELISA, the sensitivity of the test, and high accuracy, has been widely used in medical and health detection, clinical diagnosis and disease inspection, verification of harmful substances in food residues, auxiliary gene engineering etc。。 In this study, we tested the experimental effect of tumor necrosis factor alpha, interleukin -1 beta, interleukin -6 and vascular cell adhesion molecule -1 by ELISA double sandwich method, and improved it to improve it。 In the experiment, the control variable method is used to confirm the factors to be improved。 Through experiments, I first confirm that the difference between the four HRP effects is within the controllable range。 It was then confirmed that the effect of an anti pack was good and that the titer of the two antibody did not decrease。 Previous experiments have found that the speed of color rendering is too high and the cost is too high。 By using the control variable method, and setting HRP and two anti dilution multiple control experiments, the final dilution times of HRP and two resistance were determined to reduce the background。 Finally, the target of a complete cytokine testing kit is achieved。
Keyword：Double Antibody Sandwich Method; TNF-α; IL-1β; IL-6; human-VCAM-1
目录
1绪论 1
1。1。1 ELISA实验的基本原理 1
1。1。2 ELISA实验的常见类型有： 1
1。1。3 ELISA测试试剂准备 2
1。1。4 ELISA技术目前的适用范围 5
1。1。5 ELISA技术现在存在的问题 6
1。1。6 ELISA试剂盒 6
1。2 课题现状 7
1。2。1 课题背景及课题目的 7
1。2。2 测定指标介绍 8
1。2。3 检测方法详述 10
1。2。4 课题意义 11
2材料与方法 12
2。1 实验试剂的制备 12
2。2 实验仪器的选择 13
2。3 实验方案的设计 13
2。4 实验过程 14
3 结果与分析 17
3。1 验证四支HRP效果差异