基于马赫-曾德尔干涉的免疫传感器用于检测牛奶、巧克力牛奶和酸奶中的黄曲霉毒素 M1。
Mach-Zehnder Interferometric Immunosensor for Detection of Aflatoxin M1 in Milk, Chocolate Milk, and Yogurt.
机构信息
Immunoassays/Immunosensors Lab, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, NCSR "Demokritos", 15341 Aghia Paraskevi, Greece.
Analytical Chemistry Lab, Department of Chemistry, University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece.
出版信息
Biosensors (Basel). 2023 May 30;13(6):592. doi: 10.3390/bios13060592.
Aflatoxin M1 (AFM1) is detected in the milk of animals after ingestion of aflatoxin B1-contaminated food; since 2002, it has been categorized as a group I carcinogen. In this work, a silicon-based optoelectronic immunosensor for the detection of AFM1 in milk, chocolate milk, and yogurt has been developed. The immunosensor consists of ten Mach-Zehnder silicon nitride waveguide interferometers (MZIs) integrated on the same chip with the respective light sources, and an external spectrophotometer for transmission spectra collection. The sensing arm windows of MZIs are bio-functionalized after chip activation with aminosilane by spotting an AFM1 conjugate with bovine serum albumin. For AFM1 detection, a three-step competitive immunoassay is employed, including the primary reaction with a rabbit polyclonal anti-AFM1 antibody, followed by biotinylated donkey polyclonal anti-rabbit IgG antibody and streptavidin. The assay duration was 15 min with limits of detection of 0.005 ng/mL in both full-fat and chocolate milk, and 0.01 ng/mL in yogurt, which are lower than the maximum allowable concentration of 0.05 ng/mL set by the European Union. The assay is accurate (% recovery values 86.7-115) and repeatable (inter- and intra-assay variation coefficients <8%). The excellent analytical performance of the proposed immunosensor paves the way for accurate on-site AFM1 determination in milk.
黄曲霉毒素 M1(AFM1)是动物摄入受黄曲霉毒素 B1 污染的食物后在其奶中被检测到的物质;自 2002 年以来,它被归类为 I 类致癌物。在这项工作中,开发了一种用于检测牛奶、巧克力奶和酸奶中 AFM1 的基于硅的光电免疫传感器。该免疫传感器由十个硅氮化物波导马赫-曾德尔干涉仪(MZI)组成,这些干涉仪与各自的光源一起集成在同一芯片上,并且有一个外部分光光度计用于传输光谱采集。MZI 的传感臂窗口在芯片用氨丙基硅烷激活后通过用牛血清白蛋白标记的 AFM1 缀合物进行点样而被生物功能化。用于 AFM1 检测的是三步竞争免疫测定法,包括与兔多克隆抗 AFM1 抗体的初次反应,接着是生物素化驴多克隆抗兔 IgG 抗体和链霉亲和素。测定持续时间为 15 分钟,全脂奶和巧克力奶中的检测限为 0.005ng/mL,酸奶中的检测限为 0.01ng/mL,均低于欧盟设定的最大允许浓度 0.05ng/mL。该测定法准确(回收率值为 86.7-115%)且可重复(批内和批间变异系数 <8%)。所提出的免疫传感器具有出色的分析性能,为在现场准确测定奶中的 AFM1 铺平了道路。