Development of a test strip for rapid detection of lactoperoxidase in raw milk  

乳过氧化物酶快速检测试纸的研制（英文）

作　　者：Hong-xia CHE Bo TIAN Li-na BAI Li-ming CHENG Li-li LIU Xiao-na ZHANG Zhan-mei JIANG Xiao-xi XU 

机构地区：[1]Key Laboratory of Dairy Science (KLDS), Ministry of Education, Northeast Agricultural University

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2015年第8期672-679,共8页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the National "Twelfth Five-Year" Plan for Science and Technology Support Program of China(No.2013BAD18B06);the Open Research Fund for the Key Laboratory of Dairy Science of Northeast Agricultural University(No.KLDS201201);the Innovative Research Team of Higher Education of Heilongjiang Province(No.2010td11),China

摘　　要：Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid （CA）/0.2 mol/L disodium hydrogen phosphate （NAP） buffer solution （pH 5.0）, 22 mmol/L 3,3＇,5,5＇-tetramethylbenzidine （TMB）, 0.6 mmol/L hydrogen peroxide （H202）, and 0.5% Tween-20 or 0.3% cetyltri- methyl ammonium bromide （CTAB） were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation （CV） of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation （c=0.9776）. So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.目的：开发快速、灵敏和准确的乳过氧化物酶检测方法,用于原料乳验收及相关乳制品加工中对乳过氧化物酶的检测。创新点：首次以试纸检测方法代替传统的试管检测法,借助便携式光电测色仪实现对乳过氧化物酶的定量检测分析。方法：以反光值为检测指标,从缓冲液p H、四甲基联苯胺（TMB）浓度、过氧化氢（H2O2）浓度和表面活性剂四个方面,优化筛选检测试纸的最佳制作条件;通过精密度试验、最低检出浓度和与试管检测法相关性分析,评价试纸检测法的可靠性和准确性。结论：试纸检测体系的优化结果为0.1 mol/L柠檬酸/0.2 mol/L磷酸氢二钠溶液（p H 5.0）,22 mmol/L TMB,0.5%Tween-20或0.3%溴化十六烷基三甲铵（CTAB）,和0.6 mmol/L H2O2。检测试纸的精密度较好,变异系数在2.47%~6.72%（表1）;最低检出浓度为1.000~2.000 mg/L（图7）;与试管法检测结果的相关系数为0.9776（图8）。因此,试纸检测法是一种快速、灵敏和准确的乳过氧化物酶检测技术,可满足原料乳验收及相关乳制品加工中对乳过氧化物酶检测的需求。

关 键 词：LACTOPEROXIDASE Test strip Rapid detection Raw milk 3 3＇ 5 5＇-Tetramethylbenzidine （TMB） 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]