This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters rangi...This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters ranging from 50μm to 5μm,and these partitions are organized into six distinct blocks to facilitate image processing.Due to the superior thermal conductivity of Si and its potential for mass production,the dPCR chips were fabricated on a Si substrate.A temperature control system based on a high-power density Peltier element and a preheating/cooling PCR protocol user interface shortening the thermal cycle time.The optical design employs four 470 nm light-emitting diodes as light sources,with filters and mirrors effectively managing the light emitted during PCR.An algorithm is utilized for image processing and illumination nonuniformity correction including conversion to a monochromatic format,partition identification,skew correction,and the generation of an image correction mask.We validated the device using a range of deoxyribonucleic acid targets,demonstrating its potential applicability across multiple fields.Therefore,we provide guidance and verification of the design and testing of the recently proposed SPEED device.展开更多
This study presents a rapid and versatile low-cost sample-to-answer system for SARS-CoV-2 diagnostics.The system integrates the extraction and purification of nucleic acids,followed by amplification via either reverse...This study presents a rapid and versatile low-cost sample-to-answer system for SARS-CoV-2 diagnostics.The system integrates the extraction and purification of nucleic acids,followed by amplification via either reverse transcriptionquantitative polymerase chain reaction(RT–qPCR)or reverse transcription loop-mediated isothermal amplification(RT–LAMP).By meeting diverse diagnostic and reagent needs,the platform yields testing results that closely align with those of commercial RT-LAMP and RT‒qPCR systems.Notable advantages of our system include its speed and costeffectiveness.The assay is completed within 28 min,including sample loading(5 min),ribonucleic acid(RNA)extraction(3 min),and RT-LAMP(20 min).The cost of each assay is≈$9.5,and this pricing is competitive against that of Food and Drug Administration(FDA)-approved commercial alternatives.Although some RNA loss during on-chip extraction is observed,the platform maintains a potential limit of detection lower than 297 copies.Portability makes the system particularly useful in environments where centralized laboratories are either unavailable or inconveniently located.Another key feature is the platform’s versatility,allowing users to choose between RT‒qPCR or RT‒LAMP tests based on specific requirements.展开更多
基金support from grant no.52150710541 provided by the National Natural Science Foundation of P.R.Chinagrant no.2018YFE0109000 provided by the Ministry of Science and Technology of the P.R.China+6 种基金H.Z.was supported by grant no.62301412 from the Natural Science Foundation of Chinagrant no.2023-JCQN-0130 from the Natural Science Basic Research Program of Shaanxi Province,P.R.Chinagrant no.2023M732815 from the P.R.China Postdoctoral Science Foundationgrant no.2023BSHEDZZ18 from the Postdoctoral Science Foundation of Shaanxi provinceM.K.was supported by grant no.LTACH19005 from the Ministry of Education,YouthSports of the Czech Republic and grant no.RVO-VFN 64165 from the Ministry of Health of the Czech RepublicMinistry of Health,Czech Republic-conceptual development of research organization 00064165,General University Hospital in Prague.
文摘This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters ranging from 50μm to 5μm,and these partitions are organized into six distinct blocks to facilitate image processing.Due to the superior thermal conductivity of Si and its potential for mass production,the dPCR chips were fabricated on a Si substrate.A temperature control system based on a high-power density Peltier element and a preheating/cooling PCR protocol user interface shortening the thermal cycle time.The optical design employs four 470 nm light-emitting diodes as light sources,with filters and mirrors effectively managing the light emitted during PCR.An algorithm is utilized for image processing and illumination nonuniformity correction including conversion to a monochromatic format,partition identification,skew correction,and the generation of an image correction mask.We validated the device using a range of deoxyribonucleic acid targets,demonstrating its potential applicability across multiple fields.Therefore,we provide guidance and verification of the design and testing of the recently proposed SPEED device.
基金support of Grant No.52150710541 from the National Natural Science Foundation of China(NSFC),P.R.of ChinaGrant No.2018YFE0109000 from the Ministry of Science and Technology of the P.R.of China+2 种基金Petra Vopařilováand other members of Mendel University would like to acknowledge financial support from the Internal Grant Agency of the Faculty of AgriSciences,Mendel University in Brno(No.AF-IGA2023-IP-030)Haoqing Zhang would like to acknowledge the financial support of Grant No.2023-JC-QN-0130 from the Natural Science Basic Research Program of Shaanxi Province,P.R.ChinaGrant No.2023M732815 from the P.R.China Postdoctoral Science Foundation.
文摘This study presents a rapid and versatile low-cost sample-to-answer system for SARS-CoV-2 diagnostics.The system integrates the extraction and purification of nucleic acids,followed by amplification via either reverse transcriptionquantitative polymerase chain reaction(RT–qPCR)or reverse transcription loop-mediated isothermal amplification(RT–LAMP).By meeting diverse diagnostic and reagent needs,the platform yields testing results that closely align with those of commercial RT-LAMP and RT‒qPCR systems.Notable advantages of our system include its speed and costeffectiveness.The assay is completed within 28 min,including sample loading(5 min),ribonucleic acid(RNA)extraction(3 min),and RT-LAMP(20 min).The cost of each assay is≈$9.5,and this pricing is competitive against that of Food and Drug Administration(FDA)-approved commercial alternatives.Although some RNA loss during on-chip extraction is observed,the platform maintains a potential limit of detection lower than 297 copies.Portability makes the system particularly useful in environments where centralized laboratories are either unavailable or inconveniently located.Another key feature is the platform’s versatility,allowing users to choose between RT‒qPCR or RT‒LAMP tests based on specific requirements.