Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification
Liquid assays, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), are difficult to implement in point-of-care microfluidic devices because bubbles often form. We present a reaction chamber design called same-depth inlet outlet (SDIO), which was found to reduce...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-12-01
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| Series: | Sensors and Actuators Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666053925000724 |
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| author | Vi T. Nguyen Clifford Anderson Karen S. Anderson Jennifer Blain Christen |
| author_facet | Vi T. Nguyen Clifford Anderson Karen S. Anderson Jennifer Blain Christen |
| author_sort | Vi T. Nguyen |
| collection | DOAJ |
| description | Liquid assays, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), are difficult to implement in point-of-care microfluidic devices because bubbles often form. We present a reaction chamber design called same-depth inlet outlet (SDIO), which was found to reduce the instances of bubble formation by an average of 92.2 % across different flow rates when compared to traditional designs. We designed the fabrication process using xurography, laser cutting, and lamination techniques, which enable rapid and low-cost prototyping. The fabrication methods were evaluated for nuclease contamination, and best practices to reduce nucleases during and after chip assembly are detailed. Our technique, using a combination of ethanol rinses and ultraviolet-C light radiation, was found to reduce RNases up to tenfold. Different materials were tested for microfluidic compatibility with LAMP assay reagents by making chips that realistically emulate final surface areas and volumes. A variation in performance was found among different adhesives, where the best fluorescence ratio between positive and negative reactions was 4.63. Our microfluidic design was validated by amplifying a spiked RT-LAMP assay with SARS-CoV-2 primers in the reaction chambers. The overall findings aim to facilitate early-stage development and prototyping of microfluidic devices with consistent and reliable results. |
| format | Article |
| id | doaj-art-1100da5183204dc883dd4e60dca7d25f |
| institution | Kabale University |
| issn | 2666-0539 |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Sensors and Actuators Reports |
| spelling | doaj-art-1100da5183204dc883dd4e60dca7d25f2025-08-20T04:01:00ZengElsevierSensors and Actuators Reports2666-05392025-12-011010035410.1016/j.snr.2025.100354Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplificationVi T. Nguyen0Clifford Anderson1Karen S. Anderson2Jennifer Blain Christen3Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ, USACenter for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ, USACenter for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ, USACenter for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ, USA; Corresponding author.Liquid assays, such as polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP), are difficult to implement in point-of-care microfluidic devices because bubbles often form. We present a reaction chamber design called same-depth inlet outlet (SDIO), which was found to reduce the instances of bubble formation by an average of 92.2 % across different flow rates when compared to traditional designs. We designed the fabrication process using xurography, laser cutting, and lamination techniques, which enable rapid and low-cost prototyping. The fabrication methods were evaluated for nuclease contamination, and best practices to reduce nucleases during and after chip assembly are detailed. Our technique, using a combination of ethanol rinses and ultraviolet-C light radiation, was found to reduce RNases up to tenfold. Different materials were tested for microfluidic compatibility with LAMP assay reagents by making chips that realistically emulate final surface areas and volumes. A variation in performance was found among different adhesives, where the best fluorescence ratio between positive and negative reactions was 4.63. Our microfluidic design was validated by amplifying a spiked RT-LAMP assay with SARS-CoV-2 primers in the reaction chambers. The overall findings aim to facilitate early-stage development and prototyping of microfluidic devices with consistent and reliable results.http://www.sciencedirect.com/science/article/pii/S2666053925000724MicrofluidicsPoint-of-careDiagnosticsLab-on-a-chipLAMPNucleic acid amplification |
| spellingShingle | Vi T. Nguyen Clifford Anderson Karen S. Anderson Jennifer Blain Christen Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification Sensors and Actuators Reports Microfluidics Point-of-care Diagnostics Lab-on-a-chip LAMP Nucleic acid amplification |
| title | Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification |
| title_full | Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification |
| title_fullStr | Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification |
| title_full_unstemmed | Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification |
| title_short | Design, fabrication, and decontamination of low-cost microfluidics for nucleic acid amplification |
| title_sort | design fabrication and decontamination of low cost microfluidics for nucleic acid amplification |
| topic | Microfluidics Point-of-care Diagnostics Lab-on-a-chip LAMP Nucleic acid amplification |
| url | http://www.sciencedirect.com/science/article/pii/S2666053925000724 |
| work_keys_str_mv | AT vitnguyen designfabricationanddecontaminationoflowcostmicrofluidicsfornucleicacidamplification AT cliffordanderson designfabricationanddecontaminationoflowcostmicrofluidicsfornucleicacidamplification AT karensanderson designfabricationanddecontaminationoflowcostmicrofluidicsfornucleicacidamplification AT jenniferblainchristen designfabricationanddecontaminationoflowcostmicrofluidicsfornucleicacidamplification |