[Establishment of a quantitative detection method for Golgi protein73].

中华实验和临床病毒学杂志 Pub Date : 2013-10-01
Jun Hou, Tong-Sheng Guo, Jing Zhao, Ai-Xia Liu, Yong-Ji Song, Jing-Xia Guo, Jia Liu, Lin Chen, Jun Xu, Hong-Shan Wei, Bo-An Li
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引用次数: 0

Abstract

Objective: To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.

Methods: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.

Results: The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.

Conclusion: Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.

高尔基蛋白73定量检测方法的建立
目的:建立高尔基蛋白73 (GP73)定量检测的酶联免疫吸附法(ELISA)。方法:以辣根过氧化物酶标记的GP73单克隆抗体为催化酶,进行夹心反应。对几种反应液的浓度和反应条件进行了优化。对该方法进行了线性范围、灵敏度、特异度、稳定性等评价。结果:在25 ~ 500 ng/ml范围内呈线性关系。检出限为18.5 ng/ml。组间和组内RSD均小于10%。三种不同加标浓度样品的加标回收率分别为95.3%、92.6%和103.7%。在4℃和37℃贮藏3、5、7 d后,相关系数均大于0.98,RSD均小于10%。结论:所建立的ELISA法测定血清GP73含量具有较高的准确性、灵敏度和重复性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
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