Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device
Abstract A photonic quantitative polymerase chain reaction (qPCR) has usually implemented a polydimethylsiloxane (PDMS) based disposable inexpensive PCR reactor, worked as the photothermal cycler, to show potential as a point-of-care test (PoCT) for detection nucleic acids. However, the PoCT type ph...
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BMC
2025-03-01
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| Series: | Journal of Nanobiotechnology |
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| Online Access: | https://doi.org/10.1186/s12951-025-03287-0 |
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| author | Jiyeon Han Tiara A M Seongryeong Kim Gabriela Morales Florez Kiran Shrestha Dang Du Nguyen Inki Kim Jinkee Lee Gyoujin Cho |
| author_facet | Jiyeon Han Tiara A M Seongryeong Kim Gabriela Morales Florez Kiran Shrestha Dang Du Nguyen Inki Kim Jinkee Lee Gyoujin Cho |
| author_sort | Jiyeon Han |
| collection | DOAJ |
| description | Abstract A photonic quantitative polymerase chain reaction (qPCR) has usually implemented a polydimethylsiloxane (PDMS) based disposable inexpensive PCR reactor, worked as the photothermal cycler, to show potential as a point-of-care test (PoCT) for detection nucleic acids. However, the PoCT type photonic qPCR has to overcome the prolonged time for the fabrication of PDMS-based PCR reactors and enable a rapid thermal cycler to shorten diagnosis time with a strong fluorescence intensity. Here, we developed a room-temperature curable titanium dioxide (TiO2) nanoparticle dispersed PDMS (TiO2-PDMS) nanocomposite to reduce the fabrication time of the PCR reactor which enhanced the speed of photothermal cycles and fluorescence signal intensity of photonic qPCR. The TiO2-PDMS nanocomposite was formulated for rapid cross-linking at the room-temperature by introducing an optimized amount of Pt catalyst, resulting in the fabrication of a nanocomposite-based PCR reactor within 8 min at room-temperature. The nanocomposite-based PCR reactor enhanced the heating rate to 18.33 Cº/s and cooling rate to −3.11Cº/s because of the phonon scattering effect of TiO2 in the reactor and successfully amplified λ-DNA (amplicon size of 100 bp) within 10 min. Finally, we improved the qPCR efficiency by 2 cycle threshold (C t ) value compared with pristine PDMS reactor and quantified up to 10 copies/µL nucleic acids by fluorescence intensity enhancement resulting from light reflections property of TiO2. By using TiO2-PDMS nanocomposite-based PCR reactors, the fast and efficient nucleic acid assay was enabled without loss of sensitivity, and it can be practically used in the field of PoCT. Graphical Abstract |
| format | Article |
| id | doaj-art-989b59a25ddd4c4fbbd76b4c5c3e8349 |
| institution | Kabale University |
| issn | 1477-3155 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj-art-989b59a25ddd4c4fbbd76b4c5c3e83492025-08-20T03:40:47ZengBMCJournal of Nanobiotechnology1477-31552025-03-0123111310.1186/s12951-025-03287-0Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR deviceJiyeon Han0Tiara A M1Seongryeong Kim2Gabriela Morales Florez3Kiran Shrestha4Dang Du Nguyen5Inki Kim6Jinkee Lee7Gyoujin Cho8Department of Biophysics, Sungkyunkwan UniversityDepartment of Biophysics, Sungkyunkwan UniversityDepartment of Biophysics, Sungkyunkwan UniversityDepartment of Intelligent Precision Healthcare Convergence, Sungkyunkwan UniversityInstitute of Quantum Biophysics, Sungkyunkwan UniversityDepartment of Biophysics, Sungkyunkwan UniversityDepartment of Biophysics, Sungkyunkwan UniversityInstitute of Quantum Biophysics, Sungkyunkwan UniversityDepartment of Biophysics, Sungkyunkwan UniversityAbstract A photonic quantitative polymerase chain reaction (qPCR) has usually implemented a polydimethylsiloxane (PDMS) based disposable inexpensive PCR reactor, worked as the photothermal cycler, to show potential as a point-of-care test (PoCT) for detection nucleic acids. However, the PoCT type photonic qPCR has to overcome the prolonged time for the fabrication of PDMS-based PCR reactors and enable a rapid thermal cycler to shorten diagnosis time with a strong fluorescence intensity. Here, we developed a room-temperature curable titanium dioxide (TiO2) nanoparticle dispersed PDMS (TiO2-PDMS) nanocomposite to reduce the fabrication time of the PCR reactor which enhanced the speed of photothermal cycles and fluorescence signal intensity of photonic qPCR. The TiO2-PDMS nanocomposite was formulated for rapid cross-linking at the room-temperature by introducing an optimized amount of Pt catalyst, resulting in the fabrication of a nanocomposite-based PCR reactor within 8 min at room-temperature. The nanocomposite-based PCR reactor enhanced the heating rate to 18.33 Cº/s and cooling rate to −3.11Cº/s because of the phonon scattering effect of TiO2 in the reactor and successfully amplified λ-DNA (amplicon size of 100 bp) within 10 min. Finally, we improved the qPCR efficiency by 2 cycle threshold (C t ) value compared with pristine PDMS reactor and quantified up to 10 copies/µL nucleic acids by fluorescence intensity enhancement resulting from light reflections property of TiO2. By using TiO2-PDMS nanocomposite-based PCR reactors, the fast and efficient nucleic acid assay was enabled without loss of sensitivity, and it can be practically used in the field of PoCT. Graphical Abstracthttps://doi.org/10.1186/s12951-025-03287-0Photonic PCRPlasmonic fluorescenceTiO2-PDMSPhotothermal cyclerNucleic acid assay |
| spellingShingle | Jiyeon Han Tiara A M Seongryeong Kim Gabriela Morales Florez Kiran Shrestha Dang Du Nguyen Inki Kim Jinkee Lee Gyoujin Cho Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device Journal of Nanobiotechnology Photonic PCR Plasmonic fluorescence TiO2-PDMS Photothermal cycler Nucleic acid assay |
| title | Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device |
| title_full | Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device |
| title_fullStr | Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device |
| title_full_unstemmed | Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device |
| title_short | Nanocomposite-based PCR reactors to enhance thermal rate and fluorescence intensity in hand-held qPCR device |
| title_sort | nanocomposite based pcr reactors to enhance thermal rate and fluorescence intensity in hand held qpcr device |
| topic | Photonic PCR Plasmonic fluorescence TiO2-PDMS Photothermal cycler Nucleic acid assay |
| url | https://doi.org/10.1186/s12951-025-03287-0 |
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