A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis

Abstract

A silicon lab-on-chip, for the detection of nucleic acids through the integrated PCR and hybridization microarray, was developed. The silicon lab-on-chip manufactured through bio-MEMS technology is composed of two PCR microreactors (each volume 11.2 µL) and a microarray-hybridization microchamber (volume 30 µL), fluidically connected by buried bypass. It contains heaters and temperature sensors for the management and control of the temperature cycles during the PCR amplification and hybridization processes. A post-silicon process based on (i) plasmo-O2 cleaning/activation, (ii) vapor phase epoxy silanization, (iii) microarray fabrication and (iv) a protein-based passivation step was developed and fully characterized. The ssDNA microarray (4 rows × 10 columns) composed of 400 spots (spot size—70 ± 12 µm; spot-to-spot distance—130 ± 13 µm) was manufactured by piezo-dispense technology. A DNA microarray probe density in the range of 1310 to 2070 probe µm−2 was observed, together with a limit of detection of about 19 target µm−2. The performances of the silicon lab-on-chip were validated by the detection of the beta-globin gene directly from human blood. Remarkable sensitivity, multiplexing analysis and specificity were demonstrated for the detection of beta-globin and mycobacterium tuberculosis sequences.

A Short Review on Miniaturized Biosensors for the Detection of Nucleic Acid Biomarkers

“…Ventimiglia et al [ 117 ] created a silicon lab-on-chip to detect nucleic acids using a hybridization microarray and integrated PCR. A couple of PCR microdevices with a volume capability of 11.2 L and a microarray-hybridization microchamber with a volume of 30 L, which are fluidically united by buried bypass and made up of silicon LoC, were created using bio-MEMS technology ( Figure 5 B).…”

“… ( A ) Overview of the process and microscopic cross section of channel and electrodes [ 116 ] ( B ) Schematic of the LoC developed by STMicroelectronics [ 117 ] ( C ) Schematic of meta surface biosensors [ 118 ] ( D ) Schematic of dielectric meta surface structure and the whole view of meta surface substrate [ 119 ]. …”

Smart Sensors and Microtechnologies in the Precision Medicine Approach against Lung Cancer

“…The amplification system consists of a micro-chamber reactor with a hybrid silicon-polymer structure and the temperature control systems are implemented with microheaters integrated on a silicon substrate. For instance, on the silicon microsystem, the process for grafting DNA probes to electrodes can occur through electrochemical polymerization of a pyrrole group [ 189 ] or highly performant composition of two PCR microreactors and a microarray hybridization microchamber, fluidically connected by buried bypass [ 190 ]. In addition, [ 117 ].…”

Functionalized DNA Origami‐Enabled Detection of Biomarkers

“…[28,29] Traditional detection methods such as real-time quantitative PCR (qPCR), microarray hybridization, RNA sequencing, and Northern blotting, are effective but can be complex, costly, and timeconsuming. [30][31][32][33][34][35][36][37] The advent of DNA origami platforms has marked a significant advancement in the detection of nucleic acid biomarkers. This innovative approach facilitates the integration of multiple binding sites, allowing for the concurrent capture of various target miRNAs at the nanoscale.…”

“…Assessing their presence, quantity, mutations, and expression levels helps healthcare providers to provide accurate diagnoses, customize treatment plans, and monitor disease progression [28,29] . Traditional detection methods such as real‐time quantitative PCR (qPCR), microarray hybridization, RNA sequencing, and Northern blotting, are effective but can be complex, costly, and time‐consuming [30–37] . The advent of DNA origami platforms has marked a significant advancement in the detection of nucleic acid biomarkers.…”