背景荧光猝灭免疫层析测定牛奶中林可霉素

Test of lincomycin in milk based on background fluorescence quenching immune chromatography

目的:采用背景荧光猝灭-免疫层析法现场快速检测牛奶中林可霉素。方法:采用安装有Simple 1软件系统的背景荧光猝灭免疫分析仪,包括检测卡体系和背景荧光猝灭免疫分析仪,检测卡是其核心部件。基于背景荧光猝灭免疫层析法,建立牛奶中林可霉素的检测方法,制备了可用于快速检测的检测卡,同时进行方法学验证和样品测定。结果:确定检测卡最佳加样量为100μL,最佳反应时间为10 min;检测卡的灵敏度为0.088 ng·mL^-1,线性范围曲线方程Y=-5.000×10^-6X^3+0.000 4X^2+0.001 6X+0.489 2,林可霉素在0.0~60 ng·mL^-1之间呈良好的相关性(r=0.992 6)。检测卡批间差RSD均小于2%;长期重复性试验RSD均小于1%。采用本方法实测牛奶样本,与国标方法检测结果比较,t检验结果 P>0.05,说明采用背景荧光猝灭-免疫层析法和国标法2种方法比较没有显著性差异。结论:本法研制的检测卡专属性强,灵敏度高,在现场10 min内即可快速定量测定牛奶中林可霉素的含量,可应用于生鲜牛奶的质量控制及现场检测。

Objective:To establish a method of background fluorescence quenching immune chromatography(bFQIC) to determine lincomycin in milk. Methods:The instrument system of background fluorescence quenching immunochromatographic assay(bFQICA) used in this study included the test card system and the background fluorescence quenching immunoassay analyzer(Simple 1),The detection card was the core component.The detection method of lincomycin in milk was established based on background fluorescence quenching and immunochromatography,and a test card was developed for rapid detection,methodology validation and sample determination were performed simultaneously. Results:The best amount of sample quantity and react timing were 100 μL and 10 min. The sensitivity of the card was 0.088 ng·mL^-1,the linear equation was Y=-5.000×10^-6 X^3+0.000 4 X^2+0.001 6 X+0.489 2(r=0.992 6),and the linear concentration range was 0.0-60 ng·mL^-1,the RSD was lower than 2% and the RSDs of long-term repetitive experiment were lower than 1%. t test of the results obtained from the proposed method and the international test method gave a significance of P>0.05,which illustrated that there was no difference between the methods. Conclusion:The test card developed in this study is specific and sensitive,which can be used for the rapid quantitative determination of lincomycin content in fresh milk within 10 minutes in the field,and can be used for the quality control and field detection of fresh milk.