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Dr Patrick Hardinge
Cymrawd Ymchwil
Ysgol y Biowyddorau
- HardingeP@caerdydd.ac.uk
- Adeilad Syr Martin Evans, Ystafell Ystafell W/3.12, Rhodfa'r Amgueddfa, Caerdydd, CF10 3AX
Trosolwyg
Fy mhrif ddiddordeb yw canfod symiau olrhain o DNA mewn samplau biolegol. Mae hyn yn deillio o'r blynyddoedd a dreuliais yn gweithio fel Gwyddonydd Fforensig ac yn dilyn egwyddor sylfaenol gwyddoniaeth fforensig bod 'pob cyswllt yn gadael olrhain'. Rwy'n arbenigo mewn cymhwyso ymhelaethiad DNA isothermol, ymhelaethiad wedi'i gyfryngu â dolen yn bennaf (LAMP), i gynyddu'r DNA targed olrhain i lefel y gellir ei ganfod. Rwy'n gweithio'n bennaf gyda chanfod bioluminescent ac mae gennyf gydweithrediad hirsefydlog gydag ERBA MDX yn Nhrelái, Swydd Gaergrawnt ac rwy'n defnyddio ystod o dechnegau delweddu i ddelweddu a chofnodi'r canlyniadau. Rwyf hefyd wedi datblygu ac ymchwilio i ddulliau canfod fflwroleuol amgen ar gyfer ymhelaethu DNA LAMP sydd wedi'u cyhoeddi'n ddiweddar.
Mae fy mhrosiect ymchwil presennol yn fy ngalluogi i archwilio meysydd newydd gyda chydweithrediad cyffrous ym Mhrifysgol Caerdydd gyda'r Ysgolion Fferylliaeth a Pheirianneg, i ddefnyddio microhylifeg i gynhyrchu defnynnau a chelloedd artiffisial sy'n cynnwys adwaith ymhelaethu DNA bioluminescent. Mae sefydlogrwydd y defnynnau unffurf lluosog yn agor y posibilrwydd o fesur digidol LAMP ac mae'r adwaith bioluminescent yn ein galluogi i astudio'r celloedd artiffisial.
Hefyd yn y prosiect rwyf wedi datblygu sgiliau dilyniannu a biowybodeg yng nghynulliad de novo llyngyr sydd o gynyddu diddordeb milfeddygol yn y DU. Prif nod y gwaith hwn yw datblygu prawf penodol, sensitif i ganfod yr haint hwn a chyfuno'r ymchwil hon â chynhyrchu defnynnau microhylifig a chanfod mwyhad.
Cefnogir fy ymchwil gan raglen Sêr Cymru II Llywodraeth Cymru sy'n cael ei hariannu'n rhannol gan Brifysgol Caerdydd a Swyddfa Cyllid Ewropeaidd Cymru (WEFO) o dan Gronfa Datblygu Rhanbarthol Ewrop (ERDF).
Cyhoeddiad
2023
- Nan, X. et al. 2023. VarLOCK: sequencing-independent, rapid detection of SARS-CoV-2 variants of concern for point-of-care testing, qPCR pipelines and national wastewater surveillance. Scientific Reports 13(1), article number: 20832. (10.1038/s41598-023-47289-0)
- Hardinge, P. 2023. Molecular beacons - loop-mediated amplification (MB-LAMP). In: Shavrukov, Y. ed. Plant Genotyping: Methods and Protocols., Vol. 2638. Methods in Molecular Biology New York, US: Springer, pp. 289-299., (10.1007/978-1-0716-3024-2_20)
2022
- Hardinge, P. 2022. Optimized loop-mediated isothermal amplification (lamp) allows single copy detection using bioluminescent assay in real time (bart). In: Bioluminescence: Methods and Protocols., Vol. 2524. Springer, pp. 107-117., (10.1007/978-1-0716-2453-1_8)
- Nan, X. et al. 2022. VarLOCK - sequencing independent, rapid detection of SARS-CoV-2 variants of concern for point-of-care testing, qPCR pipelines and national wastewater surveillance. [Online]. medRxix: (10.1101/2022.01.06.21268555) Available at: https://www.medrxiv.org/content/10.1101/2022.01.06.21268555v1
2021
- Hardinge, P., Baxani, D. K., McCloy, T., Murray, J. A. H. and Castell, O. K. 2021. Author Correction: Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells. Scientific Reports 11(1), article number: 10748. (10.1038/s41598-021-90099-5)
2020
- Hardinge, P., Baxani, D. K., McCloy, T., Murray, J. A. H. and Castell, O. K. 2020. Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells. Scientific Reports 10(1), article number: 21886. (10.1038/s41598-020-78996-7)
- Hardinge, P. and Murray, J. A. H. 2020. Full dynamic range quantification using loop-mediated amplification (LAMP) by combining analysis of amplification timing and variance between replicates at low copy number. Scientific Reports 10, article number: 916. (10.1038/s41598-020-57473-1)
2019
- Hardinge, P. and Murray, J. A. H. 2019. Lack of specificity associated with using molecular beacons in loop mediated amplification assays. BMC Biotechnology 19, article number: 55. (10.1186/s12896-019-0549-z)
- Hardinge, P. and Murray, J. A. H. 2019. Reduced false positives and improved reporting of loop-mediated isothermal amplification using quenched fluorescent primers. Scientific Reports 9, article number: 7400. (10.1038/s41598-019-43817-z)
2018
- Hardinge, P., Kiddle, G., Tisi, L. and Murray, J. A. H. 2018. Optimised LAMP allows single copy detection of 35Sp and NOSt in transgenic maize using Bioluminescent Assay in Real Time (BART). Scientific Reports 8, article number: 17590. (10.1038/s41598-018-36207-4)
2014
- Hardinge, P. 2014. Low copy number quantification of DNA utilising Loop-mediated Amplification (LAMP) with Bioluminescent Assay in Real-Time (BART) reporter. PhD Thesis, Cardiff University.
2013
- Hardinge, P., Allard, J., Wain, A. and Watson, S. 2013. Optimisation of choline testing using Florence Iodine reagent, including comparative sensitivity and specificity with PSA and AP tests. Science & Justice 53(1), pp. 34-40. (10.1016/j.scijus.2012.03.004)
2012
- Kiddle, G. et al. 2012. GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnology 12, article number: 15. (10.1186/1472-6750-12-15)
Adrannau llyfrau
- Hardinge, P. 2023. Molecular beacons - loop-mediated amplification (MB-LAMP). In: Shavrukov, Y. ed. Plant Genotyping: Methods and Protocols., Vol. 2638. Methods in Molecular Biology New York, US: Springer, pp. 289-299., (10.1007/978-1-0716-3024-2_20)
- Hardinge, P. 2022. Optimized loop-mediated isothermal amplification (lamp) allows single copy detection using bioluminescent assay in real time (bart). In: Bioluminescence: Methods and Protocols., Vol. 2524. Springer, pp. 107-117., (10.1007/978-1-0716-2453-1_8)
Erthyglau
- Nan, X. et al. 2023. VarLOCK: sequencing-independent, rapid detection of SARS-CoV-2 variants of concern for point-of-care testing, qPCR pipelines and national wastewater surveillance. Scientific Reports 13(1), article number: 20832. (10.1038/s41598-023-47289-0)
- Hardinge, P., Baxani, D. K., McCloy, T., Murray, J. A. H. and Castell, O. K. 2021. Author Correction: Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells. Scientific Reports 11(1), article number: 10748. (10.1038/s41598-021-90099-5)
- Hardinge, P., Baxani, D. K., McCloy, T., Murray, J. A. H. and Castell, O. K. 2020. Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells. Scientific Reports 10(1), article number: 21886. (10.1038/s41598-020-78996-7)
- Hardinge, P. and Murray, J. A. H. 2020. Full dynamic range quantification using loop-mediated amplification (LAMP) by combining analysis of amplification timing and variance between replicates at low copy number. Scientific Reports 10, article number: 916. (10.1038/s41598-020-57473-1)
- Hardinge, P. and Murray, J. A. H. 2019. Lack of specificity associated with using molecular beacons in loop mediated amplification assays. BMC Biotechnology 19, article number: 55. (10.1186/s12896-019-0549-z)
- Hardinge, P. and Murray, J. A. H. 2019. Reduced false positives and improved reporting of loop-mediated isothermal amplification using quenched fluorescent primers. Scientific Reports 9, article number: 7400. (10.1038/s41598-019-43817-z)
- Hardinge, P., Kiddle, G., Tisi, L. and Murray, J. A. H. 2018. Optimised LAMP allows single copy detection of 35Sp and NOSt in transgenic maize using Bioluminescent Assay in Real Time (BART). Scientific Reports 8, article number: 17590. (10.1038/s41598-018-36207-4)
- Hardinge, P., Allard, J., Wain, A. and Watson, S. 2013. Optimisation of choline testing using Florence Iodine reagent, including comparative sensitivity and specificity with PSA and AP tests. Science & Justice 53(1), pp. 34-40. (10.1016/j.scijus.2012.03.004)
- Kiddle, G. et al. 2012. GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnology 12, article number: 15. (10.1186/1472-6750-12-15)
Gosodiad
- Hardinge, P. 2014. Low copy number quantification of DNA utilising Loop-mediated Amplification (LAMP) with Bioluminescent Assay in Real-Time (BART) reporter. PhD Thesis, Cardiff University.
Gwefannau
- Nan, X. et al. 2022. VarLOCK - sequencing independent, rapid detection of SARS-CoV-2 variants of concern for point-of-care testing, qPCR pipelines and national wastewater surveillance. [Online]. medRxix: (10.1101/2022.01.06.21268555) Available at: https://www.medrxiv.org/content/10.1101/2022.01.06.21268555v1
Bywgraffiad
Cyn fy PhD ym Mhrifysgol Caerdydd, treuliais dros ddegawd yn gweithio fel Gwyddonydd Fforensig mewn ymchwil a datblygu ac ar achosion ar gyfer y system cyfiawnder troseddol yn y Gwasanaeth Gwyddoniaeth Fforensig. Yn ystod fy PhD ac ymchwil ôl-ddoethurol dilynol ym Mhrifysgol Caerdydd, canolbwyntiais ar fesur DNA gyda chanfod fflwroleuol a bioluminescent.
Prosiect Cymrodoriaeth Sêr Cymru II:
Datblygu, gwerthuso a dilysu dyfais ficrohylifig ar gyfer canfod pathogenau gan ddefnyddio ymhelaethu DNA isothermol a chanfod bioluminescent. (bioleg foleciwlaidd)
Mae'r prosiect yn cyfuno ymhelaethiad isothermol o foleciwlau DNA targed gyda'r assay bioluminescent mewn amser real (BART), mewn cydweithrediad ag ERBA MDX, gan ddefnyddio biocemeg sy'n deillio o Fireflies. Arweiniodd fy ymchwil PhD ac ôl-ddoethurol fi at ganfod a meintioli niferoedd isel iawn o foleciwlau DNA targed ac ysgogi diddordeb mewn datblygu strategaeth feintioli ystod ddeinamig llawn sy'n gallu pennu crynodiadau o isel iawn i uchel iawn gyda chywirdeb. Mae'r prosiect yn fy ngalluogi i gydweithio ag arbenigwyr o'r Ysgolion Fferylliaeth a Pheirianneg ym Mhrifysgol Caerdydd ym maes cyffrous microhylifeg. Mae ymgorffori adweithyddion bioluminescent i nifer o ddiferion unffurf bach yn rhoi data meintiol gwerthfawr i mi o ganfod golau. Datblygu dyfais syml sy'n dwyn ynghyd fanteision ymhelaethu DNA isothermol, canfod bioluminescent BART, cynhyrchu defnynnau microhylifig a chrynhoi gyda chanfod a dadansoddi golau, yw prif ffocws fy ymchwil. Mae gen i ddiddordeb hefyd mewn datblygu ensymau luciferase thermostable o fireflies ar gyfer canfod BART ac wrth ddatblygu'r canfyddiad DNA targed tuag at nifer o barasitiaid o ddiddordeb milfeddygol. Yn fwy diweddar rwyf wedi datblygu diddordeb mewn genomeg, Oxford Nanopore Technology MinION dilyniannu a biowybodeg ar gyfer cynulliad genom de novo.