一种快速精准检测各种乳品中牛乳铁蛋白含量的广适性ELISA方法  

A Versatile ELISA Method for the Rapid and Accurate Detection of Bovine Lactoferrin Content in Various Dairy Products

作　　者：张婧[1] 朱孔迪 刘传铭 刘长振[2] 王鹏杰[1] 黄家强[1] ZHANG Jing;ZHU Kongdi;LIU Chuanming;LIU Changzhen;WANG Pengjie;HUANG Jiaqiang(Department of Nutrition and Health,China Agricultural University,Beijing 100193;Medical Experimental Center,China Academy of Chinese Medical Sciences,Beijing 100080)

机构地区：[1]中国农业大学营养与健康系,北京100193 [2]中国中医科学院医学实验中心,北京100080

出　　处：《中国乳业》2025年第3期91-99,共9页China Dairy

基　　金：河北省创新能力提升计划项目(225A7102D)。

摘　　要：[目的]验证夹心法ELISA技术是否可成为包括风味发酵乳在内的各种乳制品中牛乳铁蛋白含量的精准检测方法。[方法]利用优化的夹心法ELISA检测试剂分别检测牛乳铁蛋白(LTF)标准品溶液、巴氏杀菌乳样本、加标巴氏杀菌乳样本、12种北京地区市场常见的风味发酵乳、7种加标风味发酵乳、7种混合巴氏杀菌乳的风味发酵乳中牛乳铁蛋白含量,计算加标回收率及CV值。用该检测试剂检测3种加标风味发酵乳中牛乳铁蛋白对72℃热处理的敏感度;并与牛乳铁蛋白国标检测(高效液相色谱法)方法对比检测生乳和巴氏灭菌乳。[结果]该检测试剂可检测牛乳铁蛋白(LTF)标准品溶液和巴氏杀菌乳样本中牛乳铁蛋白,且加标回收率91.7%~106.4%。经检测,12种风味发酵乳中牛乳铁蛋白含量为无;7种加标风味发酵乳的加标回收率86.3%~109.1%,且检测CV值远低于12%;7种风味发酵乳与巴氏杀菌乳混合乳中牛乳铁蛋白含量与对照组比值在92.3%~110.5%,且大多数检测CV值远低于12%;3种加标风味发酵乳中牛乳铁蛋白含量随72℃加热时间延长而降低,且下降曲线相似。与国家标准方法对比分析,夹心法ELISA方法检测生乳和巴氏杀菌乳乳铁蛋白含量的符合率分别为97.54%、111.2%,说明该检测方法的准确性达到国标标准。[结论]夹心法ELISA检测试剂可精准检测风味发酵乳中牛乳铁蛋白含量。[Objective]To verify whether the Sandwich ELISA technique can become an accurate detection method for the content of Bovin lactoferrin in various dairy products,including flavored fermented milk.[Method]Using optimized sandwich ELISA detection reagent to detect the content of Bovin lactoferrin in standard solutions,pasteurized milk samples,spiked pasteurized milk samples,12 common flavored fermented milks in the Beijing market,7 spiked flavored fermented milks,and 7 flavored fermented milks mixed with pasteurized milk,calculating the recovery rate and CV value.Using this detection reagent to test the sensitivity of milk lactoferrin to 72℃ heat treatment in 3 spiked flavored fermented milks;and comparing the detection of raw milk and pasteurized milk with the national standard detection method for milk lactoferrin.[Result]The detection reagent can detect milk lactoferrin in standard solutions and pasteurized milk samples,with a recovery rate 91.7%~106.4%.It was found that the content of milk lactoferrin in 12 flavored fermented milks was none;the recovery rate of 7 spiked flavored fermented milks was between 86.3%~109.1%,and the CV values were all far below 12%;the ratio of milk lactoferrin content in 7 flavored fermented milks mixed with pasteurized milk compared to the control group was between 92.3%~110.5%,and most CV values were far below 12%;the content of milk lactoferrin in 3 spiked flavored fermented milks decreased with the extension of heat time at 72℃,and the decline curves were similar.Through comparative analysis with the national standard method,the compliance rates of milk lactoferrin content detected by the ELISA method in raw milk and pasteurized milk were 97.54% and 111.2%,respectively,indicating that the accuracy of this detection method meets national standards.[Conclusion]The optimized sandwich ELISA detection reagent can accurately determine the content of milk lactoferrin in flavored fermented milk.

关 键 词：牛乳铁蛋白 风味发酵乳 夹心法ELISA 

分 类 号：TS2[轻工技术与工程—食品科学与工程]