A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis
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 (v...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/8/563 |
| _version_ | 1797439208622653440 |
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| author | Giorgio Ventimiglia Massimiliano Pesaturo Alastair Malcolm Salvatore Petralia |
| author_facet | Giorgio Ventimiglia Massimiliano Pesaturo Alastair Malcolm Salvatore Petralia |
| author_sort | Giorgio Ventimiglia |
| collection | DOAJ |
| description | 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<sup>−2</sup> was observed, together with a limit of detection of about 19 target µm<sup>−2</sup>. 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. |
| first_indexed | 2024-03-09T11:49:28Z |
| format | Article |
| id | doaj.art-652a64f70f7f47839eedb7d298fcb8fa |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-09T11:49:28Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-652a64f70f7f47839eedb7d298fcb8fa2023-11-30T23:16:51ZengMDPI AGBiosensors2079-63742022-07-0112856310.3390/bios12080563A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids AnalysisGiorgio Ventimiglia0Massimiliano Pesaturo1Alastair Malcolm2Salvatore Petralia3EM Microelectronic, 2074 Marin-Epagnier, SwitzerlandSTMicroelectronics SRL, 20041 Agrate Brianza, ItalyApplied Microarrays Inc., Tempe, AZ 85284, USADepartment of Drug Science and Health, University of Catania, V. S. Sofia 64, 95125 Catania, ItalyA 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<sup>−2</sup> was observed, together with a limit of detection of about 19 target µm<sup>−2</sup>. 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.https://www.mdpi.com/2079-6374/12/8/563lab-on-chipDNA microarraynucleic acid testingsilicon biosensorhybridization |
| spellingShingle | Giorgio Ventimiglia Massimiliano Pesaturo Alastair Malcolm Salvatore Petralia A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis Biosensors lab-on-chip DNA microarray nucleic acid testing silicon biosensor hybridization |
| title | A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis |
| title_full | A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis |
| title_fullStr | A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis |
| title_full_unstemmed | A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis |
| title_short | A Miniaturized Silicon Lab-on-Chip for Integrated PCR and Hybridization Microarray for High Multiplexing Nucleic Acids Analysis |
| title_sort | miniaturized silicon lab on chip for integrated pcr and hybridization microarray for high multiplexing nucleic acids analysis |
| topic | lab-on-chip DNA microarray nucleic acid testing silicon biosensor hybridization |
| url | https://www.mdpi.com/2079-6374/12/8/563 |
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