全脂牛奶中孕酮的快速流通酶免疫测定法。

Rapid flow-through enzyme immunoassay of progesterone in whole cows' milk.

作者信息

Samsonova Jeanne V, Safronova Valentina A, Osipov Alexander P

机构信息

Department of Chemical Enzymology, Chemistry Faculty, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia; National University of Science and Technology "MISiS", 119991 Moscow, Russia.

Department of Chemical Enzymology, Chemistry Faculty, M.V. Lomonosov Moscow State University, 119991 Moscow, Russia.

出版信息

Anal Biochem. 2018 Mar 15;545:43-48. doi: 10.1016/j.ab.2018.01.011. Epub 2018 Jan 31.

DOI:10.1016/j.ab.2018.01.011

PMID:29366693

Abstract

A rapid flow-through immunoassay using an enzyme (horseradish peroxidase) as a label for quantitative and semi-quantitative determination of progesterone in whole cows' milk was developed. The flow-through test device consisted of a porous nitrocellulose membrane coated with antibodies and an absorbent membrane. The substrate solution containing 3,3',5,5' -tetramethylbenzidine was used for colour visualization. The detection limit of 0.4 ng/mL P4 was obtained by this method; analysis time did not exceed 15 min. To eliminate matrix interference a simple sample preparation procedure was used. Results of analysis of whole cows' milk samples with flow-through method were in good correlation with ELISA results (R = 0.96, n = 34). The developed rapid flow-through test system showed high efficiency for the determination of progesterone level in whole cow's milk and can be used on-site for quick identification of milk samples with low and high progesterone concentration.

摘要

开发了一种快速流通免疫测定法，使用酶（辣根过氧化物酶）作为标记物，用于定量和半定量测定全脂牛奶中的孕酮。流通测试装置由涂有抗体的多孔硝酸纤维素膜和吸收膜组成。含有3,3',5,5'-四甲基联苯胺的底物溶液用于显色。该方法获得的孕酮检测限为0.4 ng/mL；分析时间不超过15分钟。为消除基质干扰，采用了简单的样品制备程序。流通法分析全脂牛奶样品的结果与酶联免疫吸附测定结果具有良好的相关性（R = 0.96，n = 34）。所开发的快速流通测试系统在测定全脂牛奶中孕酮水平方面显示出高效性，可用于现场快速鉴定孕酮浓度低和高的牛奶样品。