ELISA法测定牛奶中黄曲霉毒素M1不确定度评定

摘要

目的：本实验对酶联免疫法检测牛奶中黄曲霉毒素 M1含量的测定结果进行不确定度的评定,确保检测结果的准确性及可靠性。方法实验分析了酶联免疫法测定牛奶中黄曲霉毒素M1的不确定度的分量及其来源,并通过计算各分量的不确定度得出检测结果的合成标准不确定度。结果酶联免疫法测定牛奶中黄曲霉毒素M1的浓度为45.741 pg/mL,扩展不确定度为11.704 pg/mL,置信水平P=95%,k=2。结论不确定度的主要来源为测量的重复性、试剂盒的灵敏度、标准曲线拟合,而酶标仪测定OD值、ELISA法操作过程中导致的不确定度可以忽略不计。选用酶联免疫法检测黄曲霉毒素M1时增加平行样的测定、注意试剂盒的灵敏度、保持标准曲线的稳定性对于提高酶联免疫法检测的准确性和可靠性具有较强的实用价值。%Objective The uncertainty of the detection results of Aflatoxin M1 in milk by enzyme-linked immunoassay was evaluated, which ensured the test results accurate and reliable.Methods The uncertainty component and the source for determination of Aflatoxin M1 in milk by enzyme-linked immunoassay (ELISA) was analyzed. According to the calculation of each component of uncertainty, combined standard uncertainty of detection results was calculated. Results It showed that the content of Aflatoxin M1 in milk by ELISA was 45.741 pg/mL and the expanded uncertainty was 11.704 pg/mL(P=95%,k=2).Conclusion Main source of uncertainty included repeatability, the sensitivity and standard curve fitting. The uncertainty of determination of OD by Microplate Reader and the operation process of ELISA method could be negligible. Increasing the parallel determination, noting the sensitivity kit and maintaining the stability of the standard curve for testing Alfatoxin M1 in milk by ELISA showed a strong practical value which could improve the accuracy and reliability.