Cheubong Chehasan, Takano Eri, Kitayama Yukiya, Sunayama Hirobumi, Minamoto Kazuaki, Takeuchi Ryota, Furutani Shunsuke, Takeuchi Toshifumi
Graduate School of Engineering, Kobe University, 1-1, Rokkodai-cho, Nada-ku, Kobe, 657-8501, Japan; Department of Chemistry, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, 39 Moo 1, Khlong 6, Pathum Thani, 12110, Thailand.
Graduate School of Engineering, Kobe University, 1-1, Rokkodai-cho, Nada-ku, Kobe, 657-8501, Japan.
Biosens Bioelectron. 2021 Jan 15;172:112775. doi: 10.1016/j.bios.2020.112775. Epub 2020 Nov 1.
Pork contamination is a serious concern for the global halal food market because many manufacturers commonly use pork instead of beef to reduce production costs. In this study, a highly sensitive fluorescent molecularly imprinted polymer nanogel (F-MIP-NG)-based sensor was developed for rapid porcine serum albumin (PSA) detection to investigate pork contamination in halal meat extracts. F-MIP-NGs were prepared via molecular imprinting and conjugation with ATTO 647N as the fluorescent reporter molecule for the post-imprinting modification (PIM) and then immobilized on gold-coated sensor chips. For achieving rapid and easy measurement, the fluorescence response was measured using a custom-made liquid handling robot equipped with a fluorescence microscope. The fluorescence response increased with increasing PSA concentration. Under optimal conditions, the F-MIP-NG-based sensors exhibited high sensitivity, a detection limit of 40 pM, a linear range of 0.25-5 nM, and excellent affinity and selectivity towards PSA, compared to potentially interfering proteins. Moreover, it was more efficient to detect beef contamination in 1 wt% pork contamination compared to the real-time polymerase chain reaction. Collectively the good analytical performance, high rates of recovery in real meat extract samples, fast detection, and a low detection limit of pork contamination (0.1 wt%) indicated the potential of the proposed sensor for detecting PSA as a marker of pork contamination in halal meat samples. The proposed sensing system based on the MIPs would open a way to establish highly sensitive and rapid sensing systems (<5 min/sample) for food analysis.
猪肉污染是全球清真食品市场的一个严重问题,因为许多制造商通常使用猪肉而非牛肉来降低生产成本。在本研究中,开发了一种基于高灵敏度荧光分子印迹聚合物纳米凝胶(F-MIP-NG)的传感器,用于快速检测猪血清白蛋白(PSA),以调查清真肉类提取物中的猪肉污染情况。F-MIP-NG通过分子印迹法制备,并与作为荧光报告分子的ATTO 647N结合用于印迹后修饰(PIM),然后固定在镀金传感器芯片上。为了实现快速简便的测量,使用配备荧光显微镜的定制液体处理机器人测量荧光响应。荧光响应随PSA浓度的增加而增加。在最佳条件下,基于F-MIP-NG的传感器表现出高灵敏度、40 pM的检测限、0.25 - 5 nM的线性范围,并且与潜在干扰蛋白相比,对PSA具有优异的亲和力和选择性。此外,与实时聚合酶链反应相比,检测1 wt%猪肉污染中的牛肉污染时,该传感器更高效。总体而言,良好的分析性能、在实际肉类提取物样品中的高回收率、快速检测以及低至0.1 wt%的猪肉污染检测限表明,所提出的传感器具有检测PSA作为清真肉类样品中猪肉污染标志物的潜力。所提出的基于分子印迹聚合物的传感系统将为建立用于食品分析的高灵敏度和快速传感系统(<5分钟/样品)开辟一条道路。