降钙素原两种免疫检测方法的建立

发布时间：2018-01-07 05:26

本文关键词：降钙素原两种免疫检测方法的建立　出处：《长春理工大学》2015年硕士论文　论文类型：学位论文

【摘要】：降钙素原是有活性的降钙素的初始结构物质,通常是由甲状腺C细胞分泌的,在机体中PCT对血液钙过多的情况有应答反应。降钙素原对于病人菌血症的预测和评估疾病的严重程度上是很重要的依据。因此医学临床近年对于降钙素原的研究及体外诊断方法引起学者广泛关注。本课题建立了降钙素原半定量胶体金免疫检测和ELISA检测方法。通过试验对反应条件以及实验用耗材和试剂来进行优化,达到最佳检测效果。对检测试剂的敏感性、重复性和特异性进行评估。实验结果表明,本研究成功的建立了降钙素原半定量胶体金免疫检测方法,并且制作了标准比色卡。检测范围为10 ng/ml、2 ng/ml、0.5 ng/ml、0.1 ng/ml,显色效果和灵敏度较好。性能评估表明研究中建立的试纸条的特异性、重复性以及热稳定性均表现良好。与市售的国产试剂盒相比显色梯度明显,检测限更低,显色效果更好。成功建立了ELISA检测方法,确定了阳性样品的临界值OD450nm=0.2395,线性范围在75 ng/L-1200 ng/L之间,最低检出限为75 ng/L。性能评估显示出了本检测试剂特异性良好,批内以及批间变异系数均小于7%,精密度高。与市售的国产试剂盒相比灵敏度更高,线性更好。
[Abstract]:Procalcitonin is the initial structural substance of active calcitonin, usually secreted by thyroid C cells. In the body, PCT is responsive to the condition of hypercalcemia. Procalcitonin is an important basis for predicting and evaluating the severity of the disease in patients with bacteremia. The research and in vitro diagnosis methods have attracted much attention. In this paper, a semi-quantitative colloidal gold immunoassay and a ELISA detection method for procalcitonin have been established. The reaction conditions and the consumables and reagents used in the experiment have been studied. All right, optimization. The sensitivity, reproducibility and specificity of the reagent were evaluated. The results showed that the semi-quantitative colloidal gold immunoassay for calcitonin was successfully established. The standard colorimetric card was made and the detection range was 10 ng / ml 2 ng / ml 0. 5 ng / ml 0. 1 ng/ml. The performance evaluation showed that the specificity, reproducibility and thermal stability of the test strip were good. Compared with the domestic kit, the color gradient was obvious and the detection limit was lower. The method of ELISA detection was established successfully and the critical value of OD450nm=0.2395 of positive samples was determined. The linear range was between 75 ng/L-1200 ng/L and the lowest detection limit was 75 ng / L. The performance evaluation showed that the specificity of the reagent was good. The coefficient of variation within and between batches is less than 7, and the precision is high. Compared with the domestic kits, the sensitivity is higher and the linearity is better.
【学位授予单位】：长春理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6

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