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基于亚甲基蓝掺杂二氧化硅纳米颗粒上的病毒探针检测丙型肝炎病毒(HCV1)DNA的阻抗式基因传感器

Impedimetric genosensor for detection of hepatitis C virus (HCV1) DNA using viral probe on methylene blue doped silica nanoparticles.

作者信息

Singhal Chaitali, Ingle Aviraj, Chakraborty Dhritiman, Pn Anoop Krishna, Pundir C S, Narang Jagriti

机构信息

Amity Institute of Nanotechnology, Amity University, Noida, UP, India.

Department of Biochemistry, Maharishi Dayanand University, Rohtak, Haryana, India.

出版信息

Int J Biol Macromol. 2017 May;98:84-93. doi: 10.1016/j.ijbiomac.2017.01.093. Epub 2017 Jan 23.

Abstract

An impedimetric genosensor was fabricated for detection of hepatitis C virus (HCV) genotype 1 in serum, based on hybridization of the probe with complementary target cDNA from sample. To achieve it, probe DNA complementary to HCVgene was immobilized on the surface of methylene blue (MB) doped silica nanoparticles MB@SiNPs) modified fluorine doped tin oxide (FTO) electrode. The synthesized MB@SiNPs was characterized using scanning electron microscopy (SEM), high resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD) pattern. This modified electrode (ssDNA/MB@SiNPs/FTO) served both as a signal amplification platform (due to silica nanoparticles (SiNPs) as well as an electrochemical indicator (due to methylene blue (MB)) for the detection of the HCV DNA in patient serum sample. The genosensor was optimized and evaluated. The sensor showed a dynamic linear range 100-10 copies/mL, with a detection limit of 90 copies/mL. The sensor was applied for detection of HCV in sera of hepatitis patient and could be renewed. The half life of the sensor was 4 weeks. The MB@SiNPs/FTO electrode could be used for preparation of other gensensors also.

摘要

基于探针与样品中互补靶标cDNA的杂交,制备了一种用于检测血清中丙型肝炎病毒(HCV)1型的阻抗基因传感器。为此,将与HCV基因互补的探针DNA固定在亚甲基蓝(MB)掺杂的二氧化硅纳米颗粒(MB@SiNPs)修饰的氟掺杂氧化锡(FTO)电极表面。使用扫描电子显微镜(SEM)、高分辨率透射电子显微镜(HRTEM)和X射线衍射(XRD)图谱对合成的MB@SiNPs进行了表征。这种修饰电极(ssDNA/MB@SiNPs/FTO)既作为信号放大平台(由于二氧化硅纳米颗粒(SiNPs)),又作为电化学指示剂(由于亚甲基蓝(MB))用于检测患者血清样品中的HCV DNA。对该基因传感器进行了优化和评估。该传感器的动态线性范围为100 - 10拷贝/mL,检测限为90拷贝/mL。该传感器用于检测肝炎患者血清中的HCV,并且可以更新。该传感器的半衰期为4周。MB@SiNPs/FTO电极也可用于制备其他基因传感器。

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