Departamento de Química Analítica. Universidad Autónoma de Madrid, 28049, Madrid, Spain.
Departamento de Química Analítica. Universidad Autónoma de Madrid, 28049, Madrid, Spain; Institute for Advanced Research in Chemical Sciences (IAdChem), Universidad Autónoma de Madrid, Ciudad Universitaria de Cantoblanco, 28049, Madrid, Spain.
Biosens Bioelectron. 2021 Oct 1;189:113375. doi: 10.1016/j.bios.2021.113375. Epub 2021 May 29.
This work focuses on the combination of molybdenum disulfide (MoS) and à la carte functionalized carbon nanodots (CNDs) for the development of DNA biosensors for selective and sensitive detection of pathogens. MoS flakes prepared through liquid-phase exfoliation, serves as platform for thiolated DNA probe immobilization, while thionine functionalized carbon nanodots (Thi-CNDs) are used as electrochemical indicator of the hybridization event. Spectroscopic and electrochemical studies confirmed the interaction of Thi-CNDs with DNA. As an illustration of the pathogen biosensor functioning, DNA sequences from InIA gen of Listeria monocytogenes bacteria and open reading frame sequence (ORF1ab) of SARS-CoV-2 virus were detected and quantified with a detection limit of 67.0 fM and 1.01 pM, respectively. Given the paradigmatic selectivity of the DNA hybridization, this approach allows pathogen detection in the presence of other pathogens, demonstrated by the detection of Listeria monocytogenes in presence of Escherichia coli. We note that this design is in principle amenable to any pathogen for which the DNA has been sequenced, including other viruses and bacteria. As example of the application of the method in real samples it has been used to directly detect Listeria monocytogenes in cultures without any DNA Polymerase Chain Reaction (PCR) amplification process.
这项工作专注于二硫化钼 (MoS) 和定制功能化碳纳米点 (CND) 的结合,用于开发用于选择性和灵敏检测病原体的 DNA 生物传感器。通过液相剥离制备的 MoS 薄片用作硫醇化 DNA 探针固定化的平台,而噻吩功能化的碳纳米点 (Thi-CND) 用作杂交事件的电化学指示剂。光谱和电化学研究证实了 Thi-CND 与 DNA 的相互作用。作为病原体生物传感器功能的说明,检测到李斯特菌的 InIA 基因和 SARS-CoV-2 病毒的开放阅读框序列 (ORF1ab) 的 DNA 序列,并分别以 67.0 fM 和 1.01 pM 的检测限进行定量。鉴于 DNA 杂交的典范选择性,这种方法允许在存在其他病原体的情况下检测病原体,通过在存在大肠杆菌的情况下检测李斯特菌证明了这一点。我们注意到,这种设计原则上适用于任何已经测序的病原体,包括其他病毒和细菌。作为该方法在实际样品中应用的示例,它已被用于直接检测未经任何聚合酶链反应 (PCR) 扩增过程的培养物中的李斯特菌。