牛肉和羊肉熟肉中含量的酶联免疫吸附测定方法的建立

IF 5.2 Q1 FOOD SCIENCE & TECHNOLOGY Journal of Future Foods Pub Date : 2023-06-06 DOI:10.1016/j.jfutfo.2023.05.010
Yujing Li , Jingjing Liu , Sufang Fan , Zhao Li , Jing Zhang , Erjing Zhang , Ziran Li , Yan Zhang , Chunsheng Li
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引用次数: 0

摘要

本研究建立了一种检测牛肉和羊肉成分的酶联免疫吸附试验(ELISA),并对其性能进行了测试。采用双抗体夹心ELISA法测定捕获抗体3G5的包被浓度为1:4000,酶标记抗体2E7辣根过氧化物酶(HRP)的工作浓度为1:1000,样品孵育时间为60min,检测抗体反应时间为60min。牛肉和羊肉骨骼肌肌钙蛋白I的检测限为45mg/kg,批间和批内回收率在80.4%-115.7%之间,变异系数在13.6%以下,鸡、鸭和鱼组织成分的交叉反应率在13.4%以下。在50和100g/kg下,检测结果与聚合酶链式反应(PCR)方法一致。因此,该ELISA方法可用于肉制品中牛肉和羊肉成分的定量检测。
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Establishment of enzyme-linked immunosorbent assay for beef and lamb contents in cooked meat

In this study, an enzyme-linked immunosorbent assay (ELISA) was established to detect beef and lamb components, and its performance was tested. Double-antibody sandwich ELISA was adopted and determined a coating concentration of capture antibody 3G5 of 1:4 000, a working concentration of enzyme-labeled antibody 2E7-horseradish peroxidase (HRP) of 1:1 000, a sample incubation time of 60 min and a detection antibody reaction time of 60 min. The specificity, sensitivity, repeatability and stability of this assay were determined. The limit of detection for beef and lamb skeletal muscle troponin I was 45 mg/kg, the inter-assay and intra-assay recovery rates ranged from 80.4% to 115.7%, the coefficients of variation were below 13.6%, and the cross reaction rates of the tissue components of chicken, duck and fish were below 13.4%. The sandwich ELISA method established in this study is stable and has high accuracy. The test results were consistent with the polymerase chain reaction (PCR) method at 50 and 100 g/kg. Therefore, this ELISA method can be used to quantitatively detect beef and lamb components in meat products.

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