Chen Ruipeng, Wang Hui, Zhao Yiguang, Nan Xuemei, Wei Wensong, Du Chunmei, Zhang Fan, Luo Qingyao, Yang Liang, Xiong Benhai
State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Institude of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Nanomaterials (Basel). 2022 Mar 26;12(7):1091. doi: 10.3390/nano12071091.
Interleukin-6 (IL-6) is generally used as a biomarker for the evaluation of inflammatory infection in humans and animals. However, there is no approach for the on-site and rapid detection of IL-6 for the monitoring of mastitis in dairy farm scenarios. A rapid and highly sensitive surface enhanced Raman scattering (SERS) immunofiltration assay (IFA) for IL-6 detection was developed in the present study. In this assay, a high sensitivity gold core silver shell SERS nanotag with Raman molecule 4-mercaptobenzoic acid (4-MBA) embedded into the gap was fabricated for labelling. Through the immuno-specific combination of the antigen and antibody, antibody conjugated SERS nanotags were captured on the test zone, which facilitated the SERS measurement. The quantitation of IL-6 was performed by the readout Raman signal in the test region. The results showed that the detection limit (LOD) of IL-6 in milk was 0.35 pg mL, which was far below the threshold value of 254.32 pg mL. The recovery of the spiking experiment was 87.0-102.7%, with coefficients of variation below 9.0% demonstrating high assay accuracy and precision. We believe the immunosensor developed in the current study could be a promising tool for the rapid assessment of mastitis by detecting milk IL-6 in dairy cows. Moreover, this versatile immunosensor could also be applied for the detection of a wide range of analytes in dairy cow healthy monitoring.
白细胞介素-6（IL-6）通常用作评估人和动物炎症感染的生物标志物。然而，在奶牛场场景中，尚无用于现场快速检测IL-6以监测乳腺炎的方法。本研究开发了一种用于检测IL-6的快速且高灵敏度的表面增强拉曼散射（SERS）免疫过滤分析方法（IFA）。在该分析方法中，制备了一种高灵敏度的金核银壳SERS纳米标签，其间隙中嵌入了拉曼分子4-巯基苯甲酸（4-MBA）用于标记。通过抗原与抗体的免疫特异性结合，将抗体偶联的SERS纳米标签捕获在测试区域，这便于进行SERS测量。通过读取测试区域的拉曼信号对IL-6进行定量。结果表明，牛奶中IL-6的检测限（LOD）为0.35 pg/mL，远低于254.32 pg/mL的阈值。加标实验的回收率为87.0 - 102.7%，变异系数低于9.0%，表明该分析方法具有高准确性和精密度。我们认为，本研究中开发的免疫传感器可能是通过检测奶牛乳汁中的IL-6快速评估乳腺炎的一种有前途的工具。此外，这种通用的免疫传感器还可应用于奶牛健康监测中多种分析物的检测。
