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通过硅光子免疫传感器快速检测马苏里拉奶酪和羊乳干酪掺牛乳制品。

Rapid detection of mozzarella and feta cheese adulteration with cow milk through a silicon photonic immunosensor.

机构信息

Immunoassays-Immunosensors Lab, Institute of Nuclear &Radiological Sciences & Technology, Energy & Safety, NCSR "Demokritos", Aghia Paraskevi 15341, Greece.

出版信息

Analyst. 2021 Jan 21;146(2):529-537. doi: 10.1039/d0an01706j. Epub 2020 Nov 12.

Abstract

Mozzarella di Bufala Campana and Feta are two cheeses with Protected Designation of Origin the fraudulent adulteration of which with bovine milk must be routinely checked to ensure that consumers actually buy these high-end products and avoid health issues related to bovine milk allergy. Here, we employed, for the first time, a silicon-based photonic immunosensor for the detection of mozzarella and feta adulteration with bovine milk. The photonic immunosensor used relies on Mach-Zehnder interferometers monolithically integrated along with their respective light sources on a silicon chip. A rabbit polyclonal antiserum raised against bovine κ-casein was used for the development of a competitive immunoassay realized in three steps, including a reaction with the antiserum, a biotinylated anti-rabbit IgG antibody, and streptavidin. The implementation of this assay configuration significantly reduced the non-specific signal due to the cheese matrix, and allowed completion of the assay in ∼9 min. After optimization of all assay conditions, bovine cheese could be quantified in mozzarella or feta at concentrations as low as 0.5 and 0.25% (w/w), respectively; both quantification limits were below the maximum allowable content of bovine milk in mozzarella and feta (1% w/w) according to the EU regulations. Equally important, the assays were reproducible with intra- and inter-assay coefficients of variation <10%, and exhibited a wide linear dynamic range that extended up to 50 and 25% (w/w) for mozzarella and feta, respectively. Taking into account its performance, the proposed immunosensor may be transformed to a new tool against fraudulent activities in the dairy industry.

摘要

马苏里拉干酪和菲达奶酪是两种具有受保护原产地名称的奶酪,其掺假行为必须定期进行欺诈性掺假检查,以确保消费者实际购买这些高端产品,并避免与牛奶过敏相关的健康问题。在这里,我们首次使用基于硅的光子免疫传感器来检测马苏里拉奶酪和菲达奶酪中掺有牛奶。所使用的光子免疫传感器依赖于沿硅芯片集成的马赫-曾德尔干涉仪及其各自的光源。针对牛κ-酪蛋白的兔多克隆抗血清用于开发竞争性免疫分析,该分析分三步进行,包括与抗血清、生物素化抗兔 IgG 抗体和链霉亲和素的反应。该分析配置的实施显著降低了奶酪基质引起的非特异性信号,并允许在约 9 分钟内完成分析。在优化所有分析条件后,可以在马苏里拉奶酪或菲达奶酪中定量检测低至 0.5%和 0.25%(w/w)的牛奶,这两个定量限均低于欧盟法规规定的马苏里拉奶酪和菲达奶酪中牛奶的最大允许含量(1%w/w)。同样重要的是,该分析具有<10%的批内和批间变异系数,并且表现出宽的线性动态范围,可扩展至马苏里拉奶酪和菲达奶酪的 50%和 25%(w/w)。考虑到其性能,所提出的免疫传感器可以转化为一种新的工具,以打击乳制品行业的欺诈活动。

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