African swine fever(ASF)is a highly lethal disease of domestic and wild swine caused by African swine fever virus(ASFV).The disease currently circulates in Africa,Europe,Asia and on the island of Hispaniola.The ongoin...African swine fever(ASF)is a highly lethal disease of domestic and wild swine caused by African swine fever virus(ASFV).The disease currently circulates in Africa,Europe,Asia and on the island of Hispaniola.The ongoing epizootics in Europe and Asia have produced millions of animal deaths and severe economic losses.No effective vaccine is available for ASF,making rapid and accurate detection of ASFV essential for disease mitigation strategies.Currently available diagnostics for ASFV possess significant limitations related to assay performance,deployability,and/or turn-around time;therefore there is an unmet need for pen-side diagnostic tests with sufficient sensitivity and specificity.A chromatographic lateral flow immunoassay(LFIA)was developed for the detection of ASFV antigen in EDTA-treated whole blood using monoclonal antibodies targeting the viral p30 protein.The assay requires only water to perform and provides results in 25 min,making it well-suited for field use.The LFIA was capable of detecting genotype I and genotype II strains of ASFV in EDTA blood from experimentally infected pigs at varying time-points after infection,though it was unable to detect a genotype X ASFV strain.Diagnostic sensitivity correlated with clinical disease severity,body temperature,and viral DNA levels,and was over 90%in animals showing moderate to severe ASF-related symptoms after challenge with virulent genotype II virus.The LFIA also showed a robust diagnostic specificity of over 98%,which is essential to field testing for a high consequence to foregin animal disease.The LFIA targeting the viral p30 protein can reliably detect ASFV in whole blood from animals showing moderate to severe clinical signs of infection with virulent genotype I and II isolates,making it a promising candidate for use as a field-deployable antigen detection assay.Additional evaluation using field samples and different virus strains is required to further assess the utility of this rapid diagnostic test.展开更多
Nucleic acid amplification tests(NAAT)have long been used in laboratory facilities and recently revolutionized the field of molecular diagnostics in point-of-care testing.Digital microfluidics(DMF)has emerged as a pro...Nucleic acid amplification tests(NAAT)have long been used in laboratory facilities and recently revolutionized the field of molecular diagnostics in point-of-care testing.Digital microfluidics(DMF)has emerged as a promising tool to complete the entire NAAT workflow in a miniaturized format with minimum human intervention.Based on electric fields to manipulate independent reaction droplets,the compact DMF system could perform multiple processes simultaneously and automatically in a programmable fashion.This combination is beginning to establish powerful sample-to-answer platforms in remote or resource-limited settings.Herein,we provide a comprehensive overview of the state-of-the-art DMF technology for point-of-care NAAT.This review focused on key principles of DMF platforms and the latest trends in system integration for automated processes of nucleic acid extraction,amplification,and detection.Also,this article discusses current challenges,including control systems,scalability and throughput,as well as future prospects of DMF-based NAAT strategy for the next generation of point-of-care diagnostics.展开更多
■INTRODUCTION Many health care systems lack access to robust diagnostic equipment and therapeutic treatments.This is particularly problematic in less developed countries and in remote locations i.e.small rural areas ...■INTRODUCTION Many health care systems lack access to robust diagnostic equipment and therapeutic treatments.This is particularly problematic in less developed countries and in remote locations i.e.small rural areas or villages.Laboratories in developing countries are often sparsely distributed,under-resourced with access hindered by economical or geographical factors.1 Such circumstances make the management and control of disease outbreaks and pandemics e.g.Covid-19 very difficult and challenging.Therefore,there is a need for low cost and reliable point-of-care(POC)test systems that allow for rapid diagnosis of disease beyond laboratory settings.Reverse transcription polymerase chain reaction(RT-PCR)testing is still considered the“gold-standard”method for Covid-19 detection,however its reliance on precise and expensive thermal cyclers has restricted its use in resource limited settings and for POC applications.2−4 In addition,reports of false-negative results when the RT-PCR test was utilized for Covid-19 detection,further strengthens the case for the development of alternative test systems.展开更多
We demonstrate a handheld on-chip biosensing technology that employs plasmonic microarrays coupled with a lens-free computational imaging system towards multiplexed and high-throughput screening of biomolecular intera...We demonstrate a handheld on-chip biosensing technology that employs plasmonic microarrays coupled with a lens-free computational imaging system towards multiplexed and high-throughput screening of biomolecular interactions for point-of-care applications and resource-limited settings.This lightweight and field-portable biosensing device,weighing 60 g and 7.5 cm tall,utilizes a compact optoelectronic sensor array to record the diffraction patterns of plasmonic nanostructures under uniform illumination by a single-light emitting diode tuned to the plasmonic mode of the nanoapertures.Employing a sensitive plasmonic array design that is combined with lens-free computational imaging,we demonstrate label-free and quantitative detection of biomolecules with a protein layer thickness down to 3 nm.Integrating large-scale plasmonic microarrays,our on-chip imaging platform enables simultaneous detection of protein mono-and bilayers on the same platform over a wide range of biomolecule concentrations.In this handheld device,we also employ an iterative phase retrieval-based image reconstruction method,which offers the ability to digitally image a highly multiplexed array of sensors on the same plasmonic chip,making this approach especially suitable for high-throughput diagnostic applications in field settings.展开更多
基金Funding for this study was provided through grants from the National Bio and Agro-Defense Facility(NBAF)Transition Fund from the State of Kansas,and the AMP Core of the Center of Emerging and Zoonotic Infectious Diseases(CEZID)from National Institute of General Medical Sciences(NIGMS)under award number P20GM130448.
文摘African swine fever(ASF)is a highly lethal disease of domestic and wild swine caused by African swine fever virus(ASFV).The disease currently circulates in Africa,Europe,Asia and on the island of Hispaniola.The ongoing epizootics in Europe and Asia have produced millions of animal deaths and severe economic losses.No effective vaccine is available for ASF,making rapid and accurate detection of ASFV essential for disease mitigation strategies.Currently available diagnostics for ASFV possess significant limitations related to assay performance,deployability,and/or turn-around time;therefore there is an unmet need for pen-side diagnostic tests with sufficient sensitivity and specificity.A chromatographic lateral flow immunoassay(LFIA)was developed for the detection of ASFV antigen in EDTA-treated whole blood using monoclonal antibodies targeting the viral p30 protein.The assay requires only water to perform and provides results in 25 min,making it well-suited for field use.The LFIA was capable of detecting genotype I and genotype II strains of ASFV in EDTA blood from experimentally infected pigs at varying time-points after infection,though it was unable to detect a genotype X ASFV strain.Diagnostic sensitivity correlated with clinical disease severity,body temperature,and viral DNA levels,and was over 90%in animals showing moderate to severe ASF-related symptoms after challenge with virulent genotype II virus.The LFIA also showed a robust diagnostic specificity of over 98%,which is essential to field testing for a high consequence to foregin animal disease.The LFIA targeting the viral p30 protein can reliably detect ASFV in whole blood from animals showing moderate to severe clinical signs of infection with virulent genotype I and II isolates,making it a promising candidate for use as a field-deployable antigen detection assay.Additional evaluation using field samples and different virus strains is required to further assess the utility of this rapid diagnostic test.
基金support from The Ivan Bowen Family Foundation and the Department of Physiology and Biomedical Engineering at Mayo Clinic,Rochester MN.
文摘Nucleic acid amplification tests(NAAT)have long been used in laboratory facilities and recently revolutionized the field of molecular diagnostics in point-of-care testing.Digital microfluidics(DMF)has emerged as a promising tool to complete the entire NAAT workflow in a miniaturized format with minimum human intervention.Based on electric fields to manipulate independent reaction droplets,the compact DMF system could perform multiple processes simultaneously and automatically in a programmable fashion.This combination is beginning to establish powerful sample-to-answer platforms in remote or resource-limited settings.Herein,we provide a comprehensive overview of the state-of-the-art DMF technology for point-of-care NAAT.This review focused on key principles of DMF platforms and the latest trends in system integration for automated processes of nucleic acid extraction,amplification,and detection.Also,this article discusses current challenges,including control systems,scalability and throughput,as well as future prospects of DMF-based NAAT strategy for the next generation of point-of-care diagnostics.
文摘■INTRODUCTION Many health care systems lack access to robust diagnostic equipment and therapeutic treatments.This is particularly problematic in less developed countries and in remote locations i.e.small rural areas or villages.Laboratories in developing countries are often sparsely distributed,under-resourced with access hindered by economical or geographical factors.1 Such circumstances make the management and control of disease outbreaks and pandemics e.g.Covid-19 very difficult and challenging.Therefore,there is a need for low cost and reliable point-of-care(POC)test systems that allow for rapid diagnosis of disease beyond laboratory settings.Reverse transcription polymerase chain reaction(RT-PCR)testing is still considered the“gold-standard”method for Covid-19 detection,however its reliance on precise and expensive thermal cyclers has restricted its use in resource limited settings and for POC applications.2−4 In addition,reports of false-negative results when the RT-PCR test was utilized for Covid-19 detection,further strengthens the case for the development of alternative test systems.
基金Altug Research Group acknowledges National Science Foundation(NSF)CAREER Award,Presidential Early Career Award for Scientist and Engineers(PECASE)ECCS-0954790Office of Naval Research Young Investigator Award 11PR00755-00-P00001+1 种基金NSF Engineering Research Center on Smart Lighting EEC-0812056Massachusetts Life Sciences Center Young Investigator award and Ecole Polytechnique Federale de Lausanne.Ozcan Research Group acknowledges the support of PECASE,Army Research Office(ARO)Life Sciences Division,ARO Young Investigator Award,NSF CAREER Award,ONR Young Investigator Award and the National Institute of Health(NIH)Director’s New Innovator Award DP2OD006427 from the Office of The Director,NIH and the NSF EFRI Award.
文摘We demonstrate a handheld on-chip biosensing technology that employs plasmonic microarrays coupled with a lens-free computational imaging system towards multiplexed and high-throughput screening of biomolecular interactions for point-of-care applications and resource-limited settings.This lightweight and field-portable biosensing device,weighing 60 g and 7.5 cm tall,utilizes a compact optoelectronic sensor array to record the diffraction patterns of plasmonic nanostructures under uniform illumination by a single-light emitting diode tuned to the plasmonic mode of the nanoapertures.Employing a sensitive plasmonic array design that is combined with lens-free computational imaging,we demonstrate label-free and quantitative detection of biomolecules with a protein layer thickness down to 3 nm.Integrating large-scale plasmonic microarrays,our on-chip imaging platform enables simultaneous detection of protein mono-and bilayers on the same platform over a wide range of biomolecule concentrations.In this handheld device,we also employ an iterative phase retrieval-based image reconstruction method,which offers the ability to digitally image a highly multiplexed array of sensors on the same plasmonic chip,making this approach especially suitable for high-throughput diagnostic applications in field settings.
