Park Jeong Ah, Kim Jinmyeong, Kim Soo Min, Sohn Hiesang, Park Chulhwan, Kim Tae-Hyung, Lee Jin-Ho, Lee Min-Ho, Lee Taek
Department of Chemical Engineering, Kwangwoon University, 20, Kwangwoon-Ro, Seoul 01897, Korea.
School of Integrative Engineering, Chung-Ang University, 84, Heukseok-ro, Seoul 06974, Korea.
Materials (Basel). 2021 Jan 12;14(2):343. doi: 10.3390/ma14020343.
The outbreak of the influenza virus (H1N1) has symptoms such as coughing, fever, and a sore throat, and due to its high contagious power, it is fatal to humans. To detect H1N1 precisely, the present study proposed an electrochemical biosensor composed of a multifunctional DNA four-way junction (4WJ) and carboxyl molybdenum disulfide (carboxyl-MoS) hybrid material. The DNA 4WJ was constructed to have the hemagglutinin aptamer on the head group (recognition part); each of the two arms has four silver ions (signal amplification part), and the tail group has an amine group (anchor). This fabricated multifunctional DNA 4WJ can specifically and selectively bind to hemagglutinin. Moreover, the carboxyl-MoS provides an increase in the sensitivity of this biosensor. Carboxyl-MoS was immobilized using a linker on the electrode, followed by the immobilization of the multifunctional 4WJ on the electrode. The synthesis of carboxyl-MoS was confirmed by field emission scanning electron microscopy (FE-SEM), and the surface of the electrode was confirmed by atomic force microscopy. When H1N1 was placed in the immobilized electrode, the presence of H1N1 was confirmed by electrochemical analysis (cyclic voltammetry, electrochemical impedance spectroscopy). Through selectivity tests, it was also possible to determine whether this sensor responds specifically and selectively to H1N1. We confirmed that the biosensor showed a linear response to H1N1, and that H1N1 could be detected from 100 nM to 10 pM. Finally, clinical tests, in which hemagglutinin was diluted with human serum, showed a similar tendency to those diluted with water. This study showed that the multi-functional DNA 4WJ and carboxyl-MoS hybrid material can be applied to a electrochemical H1N1 biosensor.
甲型H1N1流感病毒的爆发会引发咳嗽、发烧和喉咙痛等症状,并且由于其高传染性,对人类是致命的。为了精确检测甲型H1N1流感病毒,本研究提出了一种由多功能DNA四链体(4WJ)和羧基二硫化钼(carboxyl-MoS)混合材料组成的电化学生物传感器。构建的DNA 4WJ在头部基团(识别部分)具有血凝素适配体;两条臂中的每一条都有四个银离子(信号放大部分),尾部基团有一个胺基(锚定基团)。这种制备的多功能DNA 4WJ可以特异性地、选择性地与血凝素结合。此外,羧基二硫化钼提高了这种生物传感器的灵敏度。通过连接剂将羧基二硫化钼固定在电极上,随后将多功能4WJ固定在电极上。通过场发射扫描电子显微镜(FE-SEM)确认了羧基二硫化钼的合成,并通过原子力显微镜确认了电极表面。将甲型H1N1流感病毒置于固定化电极中时,通过电化学分析(循环伏安法、电化学阻抗谱)确认了甲型H1N1流感病毒的存在。通过选择性测试,还可以确定该传感器是否对甲型H1N1流感病毒有特异性和选择性响应。我们证实该生物传感器对甲型H1N1流感病毒呈线性响应,并且可以检测到浓度为100 nM至10 pM的甲型H1N1流感病毒。最后,用人血清稀释血凝素的临床试验显示出与用水稀释时相似的趋势。本研究表明,多功能DNA 4WJ和羧基二硫化钼混合材料可应用于甲型H1N1流感病毒的电化学生物传感器。