Sakamoto Hiroaki, Minpou Yuma, Sawai Takayuki, Enami Yasufumi, Suye Shin-ichiro
Tenure-Track Program for Innovation Research, University of Fukui, Fukui, Japan.
Research and Education Program for Life Science, University of Fukui, Bunkyo 3-9-1, Fukui, 910-8507, Japan.
Appl Biochem Biotechnol. 2016 Feb;178(4):687-94. doi: 10.1007/s12010-015-1902-x. Epub 2015 Oct 24.
In order to minimize the damage from viral epidemics, early detection of the causative agent of a viral epidemic and prevention of its immediate spread are urgent social demands. Therefore, in this study, we evaluated the utility of a Mach-Zehnder-type optical waveguide as a sensing device for influenza virus detection. However, it is impossible to detect a 100-nm-size virus using a sol-gel optical biosensor because sol-gel glass has a pore size of only a few nanometers, which makes it impossible for the virus to diffuse into the silica thin film. In order to construct the influenza-specific Mach-Zehnder optical biosensor for influenza detection, a stable antibody immobilization method with resulting high density on the sol-gel surface is strongly required. In this study, the sol-gel glass surface was modified with amino and carboxyl groups, and an anti-H1N1/HA1 antibody was covalently immobilized using a cross-linking agent. We successfully prepared a carboxyl-modified sol-gel surface, using NHS/EDC as the cross-linker, for antibody immobilization, and confirmed the detection of influenza virus using the antibody-immobilized sol-gel glass. After treatment with a 100 μg/mL influenza virus solution for 15 min, a peak wavelength shift (~24 nm) was observed in the output light spectrum.
为了将病毒流行造成的损害降至最低,尽早检测出病毒流行的病原体并防止其迅速传播是紧迫的社会需求。因此,在本研究中,我们评估了马赫-曾德尔型光波导作为流感病毒检测传感装置的实用性。然而,使用溶胶-凝胶光学生物传感器无法检测到100纳米大小的病毒,因为溶胶-凝胶玻璃的孔径只有几纳米,这使得病毒无法扩散到二氧化硅薄膜中。为了构建用于流感检测的流感特异性马赫-曾德尔光学生物传感器,强烈需要一种在溶胶-凝胶表面具有高密度且稳定的抗体固定方法。在本研究中,用氨基和羧基对溶胶-凝胶玻璃表面进行了修饰,并使用交联剂将抗H1N1/HA1抗体共价固定。我们成功制备了用NHS/EDC作为交联剂的羧基修饰溶胶-凝胶表面用于抗体固定,并使用固定有抗体的溶胶-凝胶玻璃证实了对流感病毒的检测。在用100μg/mL流感病毒溶液处理15分钟后,在输出光谱中观察到峰值波长偏移(约24纳米)。