Kusnin Norzila, Yusof Nor Azah, Abdullah Jaafar, Sabri Suriana, Mohammad Faruq, Mustafa Shuhaimi, Ab Mutalib Nurul Asyikeen, Sato Shinobu, Takenaka Shigeori, Parmin Nor Azizah, Al-Lohedan Hamad A
Institute of Advanced Technology, Universiti Putra Malaysia 43400 Serdang Selangor Malaysia
Department of Chemistry, Faculty of Science, Universiti Putra Malaysia 43400 Serdang Selangor Malaysia.
RSC Adv. 2020 Jul 22;10(46):27336-27345. doi: 10.1039/d0ra03585h. eCollection 2020 Jul 21.
In this study, an electrochemical DNA biosensor was developed based on the fabrication of silicon nanowires/platinum nanoparticles (SiNWs/PtNPs) on a screen-printed carbon electrode (SPCE) for the detection of mitochondrial DNA (mtDNA) in food utilizing a new hybrid indicator, ferrocenylnaphthalene diimide (FND). The morphology and elemental composition of the SiNWs/PtNPs-modified SPCE was analyzed by field emission scanning electron microscopy (FESEM) combined with energy dispersive X-ray spectroscopy (EDX). Cyclic voltammetry (CV) was used to study the electrical contact between the PtNPs and the screen-printed working electrode through SiNWs, while electrochemical impedance spectroscopy (EIS) was used to measure the charge transfer resistance of the modified electrode. The results clearly showed that the SiNWs/PtNPs were successfully coated onto the electrode and the effective surface area for the SiNWs/PtNPs-modified SPCE was increased 16.8 times as compared with that of the bare SPCE. Differential pulse voltammetry used for the detection of porcine DNA with FND as an intercalator confirmed its specific binding to the double-stranded DNA (dsDNA) sequences. The developed biosensor showed a selective response towards complementary target DNA and was able to distinguish non-complementary and mismatched DNA oligonucleotides. The SiNWs/PtNPs-modified SPCE that was fortified with DNA hybridization demonstrated good linearity in the range of 3 × 10 M to 3 × 10 M ( = 0.96) with a detection limit of 2.4 × 10 M. A cross-reactivity study against various types of meat and processed food showed good reliability for porcine samples.
在本研究中,基于在丝网印刷碳电极(SPCE)上制备硅纳米线/铂纳米颗粒(SiNWs/PtNPs),开发了一种电化学DNA生物传感器,用于利用新型混合指示剂二茂铁基萘二酰亚胺(FND)检测食品中的线粒体DNA(mtDNA)。通过场发射扫描电子显微镜(FESEM)结合能量色散X射线光谱(EDX)分析了SiNWs/PtNPs修饰的SPCE的形态和元素组成。循环伏安法(CV)用于研究PtNPs与通过SiNWs的丝网印刷工作电极之间的电接触,而电化学阻抗谱(EIS)用于测量修饰电极的电荷转移电阻。结果清楚地表明,SiNWs/PtNPs成功地涂覆在电极上,与裸SPCE相比,SiNWs/PtNPs修饰的SPCE的有效表面积增加了16.8倍。以FND作为嵌入剂的差分脉冲伏安法用于检测猪DNA,证实了其与双链DNA(dsDNA)序列的特异性结合。所开发的生物传感器对互补靶DNA表现出选择性响应,并且能够区分非互补和错配的DNA寡核苷酸。通过DNA杂交强化的SiNWs/PtNPs修饰的SPCE在3×10⁻⁹ M至3×10⁻⁷ M范围内表现出良好的线性(R² = 0.96),检测限为2.4×10⁻⁹ M。针对各种肉类和加工食品的交叉反应性研究表明,猪样品具有良好的可靠性。