Abstract
Surface substrate and chemical functionalization are crucial aspects for the fabrication of the sensitive biosensor based on microarray technology. In this paper, an advanced, silicon-based substrate (A-MA) allowing enhancement of optical signal for microarray application is described. The substrate consists in a multilayer of Si/Al/SiO2 layers. The optical signal enhancement is reached by a combination of the mirror effect of Al film and the SiO2 thickness around 830 nm, which is able to reach the maximum of interference for the emission wavelength of the Cy5 fluorescent label. Moreover, SiO2 layer is suitable for the immobilization of single-strand DNA through standard silane chemistry, and probe densities of about 2000 F/µm2 are reached. The microarray is investigated in the detection of HBV (Hepatitis B Virus) pathogen with analytical samples, resulting in a dynamic linear range of 0.05–0.5 nM, a sensitivity of about 18000 a.u. nM−1, and a Limit of Detection in the range of 0.031–0.043 Nm as a function of the capture probe sequence.
Highlights
Microarrays (MAs) are one of the most powerful and widely used biotechnologies for nucleic acids detection and are relevant in the field of genomics [1]
The microarray is investigated in the detection of HBV (Hepatitis B Virus) pathogen with analytical samples, resulting in a dynamic linear range of 0.05–0.5 nM, a sensitivity of about 18000 a.u. nM−1, and a Limit of Detection in the range of 0.031–0.043 Nm as a function of the capture probe sequence
The integration of microarray technology with microfluidics, microelectronics, MEMS, and optical technologies has led to the development of modern and advanced miniaturized Lab-on-chip (LoC) devices [10,11,12]
Summary
Microarrays (MAs) are one of the most powerful and widely used biotechnologies for nucleic acids detection and are relevant in the field of genomics [1]. The integration of microarray technology with microfluidics, microelectronics, MEMS, and optical technologies has led to the development of modern and advanced miniaturized Lab-on-chip (LoC) devices [10,11,12] In this context, silicon material offers several advantages for creating technological advancements in smart LoC devices. The analytical performances of ssDNA microarrays such us Limit of Detection (LoD), dynamin range, and sensitivity are correlated with (i) the specific capture probe (sequence and length), (ii) probe density (strategy of immobilization), and (iii) the experimental hybridization protocols, they are strongly affected by the level of fluorescence emission upon the probe-to-target recognition process. An ssDNA MA based on an advanced silicon-based substrate A-MA that is able to enhance the hybridization fluorescent signal for the detection of Hepatitis B Virus (HBV) specific sequence is reported. The evaluation of signal enhancement, together with the system analytical perfomances in terms of dynamic linear range, sensitivity, LoD, and LoB (Limit of Background), are presented and discussed
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