人复制蛋白变体胶乳增强免疫检测试剂盒的研制

发布时间：2018-05-27 02:33

  本文选题：肺癌 + 复制蛋白变体　； 参考：《南京大学》2015年硕士论文

【摘要】：肺癌作为发病率和死亡率增长最快的恶性肿瘤之一,是人类健康的一大杀手。近年来环境恶化,空气污染日益严重,加上一直以来居高不下的吸烟率,肺癌的发病率正呈明显上升趋势。早期肺癌由于症状不明显,加上传统的筛查方法操作复杂,很容易被忽视,因此肺癌的早期检测成为一个迫切需要解决的问题。本实验初步建立了人复制蛋白变体(CIZ1-V)的胶乳增强免疫比浊检测方法,用于早期肺癌的检测。用制备的CIZ1-V多克隆抗体(兔抗)与0.13μm粒径的胶乳微球颗粒进行化学偶联,研制CIZ1-V试剂盒,并对试剂盒性能进行鉴定。结果表明,CIZ1-V试剂盒标准曲线呈线型,相关系数R2=0.9988；检测范围为0.20～8.00 gg/mL,灵敏度为0.11μg/mL,检测限为0.20μg/mL。试剂盒对表达纯化所得目的蛋白的回收率范围为98.92%～104.00%,平均100.91%；平均批内及批间变异系数均小于5%,符合国家检测试剂盒标准。试剂盒和Elisa方法对于表达纯化所得目的蛋白的回收率无显著差异(P≥0.05),对1L菌液的CIZ1-V表达量测定结果也无显著差异(P≥0.05)。本实验初步研制出敏感、准确、特异的CIZ1-V检测试剂盒,为早期肺癌的血清中CIZ1-V的检测打下了基础。
[Abstract]:Lung cancer, one of the fastest growing malignant tumors, is a major killer of human health. In recent years, the incidence of lung cancer is on the rise due to the deterioration of the environment, the increasing air pollution and the high smoking rate. Because the symptoms of early lung cancer are not obvious, and the traditional screening method is complex, it is easy to be ignored, so the early detection of lung cancer becomes an urgent problem to be solved. In this study, a method of latex enhanced immune turbidimetry (RIA) was established for detection of early lung cancer. The CIZ1-V polyclonal antibody (rabbit antibody) was chemically coupled with 0.13 渭 m latex microspheres to prepare the CIZ1-V kit, and the properties of the kit were evaluated. The results showed that the standard curve of CIZ1-V kit was linear, the correlation coefficient was 0.9988, the detection range was 0.20 ~ 8.00 gg-mL, the sensitivity was 0.11 渭 g / mL, the detection limit was 0.20 渭 g / mL. The recovery range of the purified protein was 98.92%, the average value was 100.91.The average coefficient of variation within and between batches was less than 5, which was in line with the national detection kit standard. There was no significant difference between the kit and Elisa method in the recovery of the purified protein (P 鈮，

本文编号：1940066