Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation
Polymerase chain reaction (PCR) is the most common method used for nucleic acid (DNA) amplification. The development of PCR-performing microfluidic reactors (μPCRs) has been of major importance, due to their crucial role in pathogen detection applications in medical diagnostics. Closed loop (CL) is...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Micromachines |
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| Online Access: | https://www.mdpi.com/2072-666X/14/1/172 |
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| author | Panagiotis Skaltsounis George Kokkoris Theodoros G. Papaioannou Angeliki Tserepi |
| author_facet | Panagiotis Skaltsounis George Kokkoris Theodoros G. Papaioannou Angeliki Tserepi |
| author_sort | Panagiotis Skaltsounis |
| collection | DOAJ |
| description | Polymerase chain reaction (PCR) is the most common method used for nucleic acid (DNA) amplification. The development of PCR-performing microfluidic reactors (μPCRs) has been of major importance, due to their crucial role in pathogen detection applications in medical diagnostics. Closed loop (CL) is an advantageous type of μPCR, which uses a circular microchannel, thus allowing the DNA sample to pass consecutively through the different temperature zones, in order to accomplish a PCR cycle. CL μPCR offers the main advantages of the traditional continuous-flow μPCR, eliminating at the same time most of the disadvantages associated with the long serpentine microchannel. In this work, the performance of three different CL μPCRs designed for fabrication on a printed circuit board (PCB) was evaluated by a computational study in terms of the residence time in each thermal zone. A 3D heat transfer model was used to calculate the temperature distribution in the microreactor, and the residence times were extracted by this distribution. The results of the computational study suggest that for the best-performing microreactor design, a PCR of 30 cycles can be achieved in less than 3 min. Subsequently, a PCB chip was fabricated based on the design that performed best in the computational study. PCB constitutes a great substrate as it allows for integrated microheaters inside the chip, permitting at the same time low-cost, reliable, reproducible, and mass-amenable fabrication. The fabricated chip, which, at the time of this writing, is the first CL μPCR chip fabricated on a PCB, was tested by measuring the temperatures on its surface with a thermal camera. These results were then compared with the ones of the computational study, in order to evaluate the reliability of the latter. The comparison of the calculated temperatures with the measured values verifies the accuracy of the developed model of the microreactor. As a result of that, a total power consumption of <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mn>1.521</mn><mrow><mo> </mo><mi mathvariant="normal">W</mi></mrow></mrow></semantics></math></inline-formula> was experimentally measured, only ~7.3% larger than the one calculated (<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mn>1.417</mn><mrow><mo> </mo><mi mathvariant="normal">W</mi></mrow></mrow></semantics></math></inline-formula>). Full validation of the realized CL μPCR chip will be demonstrated in future work. |
| first_indexed | 2024-03-09T11:40:04Z |
| format | Article |
| id | doaj.art-29212c401a48433b8f156b35bb85ffbf |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-03-09T11:40:04Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj.art-29212c401a48433b8f156b35bb85ffbf2023-11-30T23:34:11ZengMDPI AGMicromachines2072-666X2023-01-0114117210.3390/mi14010172Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal ValidationPanagiotis Skaltsounis0George Kokkoris1Theodoros G. Papaioannou2Angeliki Tserepi3Institute of Nanoscience and Nanotechnology, National Center of Scientific Research (NCSR) “Demokritos”, Patr. Gregoriou Ε’ and 27 Neapoleos Str., 15341 Aghia Paraskevi, GreeceInstitute of Nanoscience and Nanotechnology, National Center of Scientific Research (NCSR) “Demokritos”, Patr. Gregoriou Ε’ and 27 Neapoleos Str., 15341 Aghia Paraskevi, GreeceSchool of Medicine, National and Kapodistrian University of Athens (NKUA), 75 Mikras Asias Str., 11527 Athens, GreeceInstitute of Nanoscience and Nanotechnology, National Center of Scientific Research (NCSR) “Demokritos”, Patr. Gregoriou Ε’ and 27 Neapoleos Str., 15341 Aghia Paraskevi, GreecePolymerase chain reaction (PCR) is the most common method used for nucleic acid (DNA) amplification. The development of PCR-performing microfluidic reactors (μPCRs) has been of major importance, due to their crucial role in pathogen detection applications in medical diagnostics. Closed loop (CL) is an advantageous type of μPCR, which uses a circular microchannel, thus allowing the DNA sample to pass consecutively through the different temperature zones, in order to accomplish a PCR cycle. CL μPCR offers the main advantages of the traditional continuous-flow μPCR, eliminating at the same time most of the disadvantages associated with the long serpentine microchannel. In this work, the performance of three different CL μPCRs designed for fabrication on a printed circuit board (PCB) was evaluated by a computational study in terms of the residence time in each thermal zone. A 3D heat transfer model was used to calculate the temperature distribution in the microreactor, and the residence times were extracted by this distribution. The results of the computational study suggest that for the best-performing microreactor design, a PCR of 30 cycles can be achieved in less than 3 min. Subsequently, a PCB chip was fabricated based on the design that performed best in the computational study. PCB constitutes a great substrate as it allows for integrated microheaters inside the chip, permitting at the same time low-cost, reliable, reproducible, and mass-amenable fabrication. The fabricated chip, which, at the time of this writing, is the first CL μPCR chip fabricated on a PCB, was tested by measuring the temperatures on its surface with a thermal camera. These results were then compared with the ones of the computational study, in order to evaluate the reliability of the latter. The comparison of the calculated temperatures with the measured values verifies the accuracy of the developed model of the microreactor. As a result of that, a total power consumption of <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mn>1.521</mn><mrow><mo> </mo><mi mathvariant="normal">W</mi></mrow></mrow></semantics></math></inline-formula> was experimentally measured, only ~7.3% larger than the one calculated (<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><mn>1.417</mn><mrow><mo> </mo><mi mathvariant="normal">W</mi></mrow></mrow></semantics></math></inline-formula>). Full validation of the realized CL μPCR chip will be demonstrated in future work.https://www.mdpi.com/2072-666X/14/1/172polymerase chain reaction (PCR)closed loopmicroreactorlab-on-chip (LoC)printed circuit board (PCB)point of care (PoC) |
| spellingShingle | Panagiotis Skaltsounis George Kokkoris Theodoros G. Papaioannou Angeliki Tserepi Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation Micromachines polymerase chain reaction (PCR) closed loop microreactor lab-on-chip (LoC) printed circuit board (PCB) point of care (PoC) |
| title | Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation |
| title_full | Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation |
| title_fullStr | Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation |
| title_full_unstemmed | Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation |
| title_short | Closed-Loop Microreactor on PCB for Ultra-Fast DNA Amplification: Design and Thermal Validation |
| title_sort | closed loop microreactor on pcb for ultra fast dna amplification design and thermal validation |
| topic | polymerase chain reaction (PCR) closed loop microreactor lab-on-chip (LoC) printed circuit board (PCB) point of care (PoC) |
| url | https://www.mdpi.com/2072-666X/14/1/172 |
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