Background:Early diagnosis and classification of infections increase the cure rate while decreasing complications,which is significant for severe infections,especially for war surgery.However,traditional methods rely ...Background:Early diagnosis and classification of infections increase the cure rate while decreasing complications,which is significant for severe infections,especially for war surgery.However,traditional methods rely on laborious operations and bulky devices.On the other hand,point-of-care(POC)methods suffer from limited robustness and accuracy.Therefore,it is of urgent demand to develop POC devices for rapid and accurate diagnosis of infections to fulfill on-site militarized requirements.Methods:We developed a wave-shaped microfluidic chip(WMC)assisted multiplexed detection platform(WMC-MDP).WMC-MDP reduces detection time and improves repeatability through premixing of the samples and reaction of the reagents.We further combined the detection platform with the streptavidin–biotin(SA-B)amplified system to enhance the sensitivity while using chemiluminescence(CL)intensity as signal readout.We realized simultaneous detection of C-reactive protein(CRP),procalcitonin(PCT),and interleukin-6(IL-6)on the detection platform and evaluated the sensitivity,linear range,selectivity,and repeatability.Finally,we finished detecting 15 samples from volunteers and compared the results with commercial ELISA kits.Results:Detection of CRP,PCT,and IL-6 exhibited good linear relationships between CL intensities and concentrations in the range of 1.25–40μg/ml,0.4–12.8 ng/ml,and 50–1600 pg/ml,respectively.The limit of detection of CRP,PCT,and IL-6 were 0.54μg/ml,0.11 ng/ml,and 16.25 pg/ml,respectively.WMC-MDP is capable of good adequate selectivity and repeatability.The whole detection procedure takes only 22 min that meets the requirements of a POC device.Results of 15 samples from volunteers were consistent with the results detected by commercial ELISA kits.Conclusions:WMC-MDP allows simultaneous,rapid,and sensitive detection of CRP,PCT,and IL-6 with satisfactory selectivity and repeatability,requiring minimal manipulation.However,WMC-MDP takes advantage of being a microfluidic device showing the coefficients of variation less than 10%enabling WMC-MDP to be a type of point-of-care testing(POCT).Therefore,WMC-MDP provides a promising alternative to POCT of multiple biomarkers.We believe the practical application of WMC-MDP in militarized fields will revolutionize infection diagnosis for soldiers.展开更多
Coronavirus disease 2019(COVID-19)is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The gold standard method for the diagnosis of SARS-CoV-2 depends on quantitative r...Coronavirus disease 2019(COVID-19)is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The gold standard method for the diagnosis of SARS-CoV-2 depends on quantitative reverse transcription-polymerase chain reaction till now,which is time-consuming and requires expensive instrumentation,and the confirmation of variants relies on further sequencing techniques.Herein,we first proposed a robust technique-methodology of electrochemical CRISPR sensing with the advantages of rapid,highly sensitivity and specificity for the detection of SARS-CoV-2 variant.To enhance the sensing capability,gold electrodes are uniformly decorated with electro-deposited gold nanoparticles.Using DNA template identical to SARS-CoV-2 Delta spike gene sequence as model,our biosensor exhibits excellent analytical detection limit(50 fM)and high linearity(R2=0.987)over six orders of magnitude dynamic range from 100 fM to 10 nM without any nucleic-acid-amplification assays.The detection can be completed within 1 h with high stability and specificity which benefits from the CRISPR-Cas system.Furthermore,based on the wireless micro-electrochemical platform,the proposed biosensor reveals promising application ability in point-of-care testing.展开更多
Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and int...Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.展开更多
The use of magnetic nanoparticle(MNP)-labeled immunochromatography test strips(ICTSs) is very important for point-ofcare testing(POCT). However, common diagnostic methods cannot accurately analyze the weak magnetic si...The use of magnetic nanoparticle(MNP)-labeled immunochromatography test strips(ICTSs) is very important for point-ofcare testing(POCT). However, common diagnostic methods cannot accurately analyze the weak magnetic signal from ICTSs, limiting the applications of POCT. In this study, an ultrasensitive multiplex biosensor was designed to overcome the limitations of capturing and normalization of the weak magnetic signal from MNPs on ICTSs. A machine learning model for sandwich assays was constructed and used to classify weakly positive and negative samples, which significantly enhanced the specificity and sensitivity. The potential clinical application was evaluated by detecting 50 human chorionic gonadotropin(HCG) samples and 59 myocardial infarction serum samples. The quantitative range for HCG was 1–1000 mIU mL^(-1) and the ideal detection limit was 0.014 mIU mL^(-1), which was well below the clinical threshold. Quantitative detection results of multiplex cardiac markers showed good linear correlations with standard values. The proposed multiplex assay can be readily adapted for identifying other biomolecules and also be used in other applications such as environmental monitoring, food analysis, and national security.展开更多
The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effect...The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effectiveness,and adaptability while providing quantitative measurements to meet the needs of laboratory testing in a variety of healthcare settings.Based on the use of microfluidic chips,MIC requires less sample and can complete sample preparation automatically.Therefore,they can provide quantitative testing results simply using a finger prick specimen.The decreased reagent consumption and reduced form factor also help improve the accessibility and affordability of healthcare services in remote and resource-limited settings.In this article,we review recent developments of the Microchip Imaging Cytometer from the following aspects:clinical applications,microfluidic chip integration,imaging optics,and image acquisition.Following,we provide an outlook of the field and remark on promising technologies that may enable significant progress in the near future.展开更多
基金the National Natural Science Foundation of China(81902167,52075138)the Natural Science Foundation of Jiangsu Province(BK20190872).
文摘Background:Early diagnosis and classification of infections increase the cure rate while decreasing complications,which is significant for severe infections,especially for war surgery.However,traditional methods rely on laborious operations and bulky devices.On the other hand,point-of-care(POC)methods suffer from limited robustness and accuracy.Therefore,it is of urgent demand to develop POC devices for rapid and accurate diagnosis of infections to fulfill on-site militarized requirements.Methods:We developed a wave-shaped microfluidic chip(WMC)assisted multiplexed detection platform(WMC-MDP).WMC-MDP reduces detection time and improves repeatability through premixing of the samples and reaction of the reagents.We further combined the detection platform with the streptavidin–biotin(SA-B)amplified system to enhance the sensitivity while using chemiluminescence(CL)intensity as signal readout.We realized simultaneous detection of C-reactive protein(CRP),procalcitonin(PCT),and interleukin-6(IL-6)on the detection platform and evaluated the sensitivity,linear range,selectivity,and repeatability.Finally,we finished detecting 15 samples from volunteers and compared the results with commercial ELISA kits.Results:Detection of CRP,PCT,and IL-6 exhibited good linear relationships between CL intensities and concentrations in the range of 1.25–40μg/ml,0.4–12.8 ng/ml,and 50–1600 pg/ml,respectively.The limit of detection of CRP,PCT,and IL-6 were 0.54μg/ml,0.11 ng/ml,and 16.25 pg/ml,respectively.WMC-MDP is capable of good adequate selectivity and repeatability.The whole detection procedure takes only 22 min that meets the requirements of a POC device.Results of 15 samples from volunteers were consistent with the results detected by commercial ELISA kits.Conclusions:WMC-MDP allows simultaneous,rapid,and sensitive detection of CRP,PCT,and IL-6 with satisfactory selectivity and repeatability,requiring minimal manipulation.However,WMC-MDP takes advantage of being a microfluidic device showing the coefficients of variation less than 10%enabling WMC-MDP to be a type of point-of-care testing(POCT).Therefore,WMC-MDP provides a promising alternative to POCT of multiple biomarkers.We believe the practical application of WMC-MDP in militarized fields will revolutionize infection diagnosis for soldiers.
基金support from the Innovation Team Project of Department of Education of Guangdong Province(No.2018KCXTD026)Guangdong Scientific and Technological Project(2019B1515120043,2020A151501612,2021A1515220109,2022B1515020093)+1 种基金Science and Technology Innovation Commission of Shenzhen(Grant No.KCXFZ20201221173413038)Longhua District Science and Innovation Commission Project Grants of Shenzhen(JCYJ201904).
文摘Coronavirus disease 2019(COVID-19)is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The gold standard method for the diagnosis of SARS-CoV-2 depends on quantitative reverse transcription-polymerase chain reaction till now,which is time-consuming and requires expensive instrumentation,and the confirmation of variants relies on further sequencing techniques.Herein,we first proposed a robust technique-methodology of electrochemical CRISPR sensing with the advantages of rapid,highly sensitivity and specificity for the detection of SARS-CoV-2 variant.To enhance the sensing capability,gold electrodes are uniformly decorated with electro-deposited gold nanoparticles.Using DNA template identical to SARS-CoV-2 Delta spike gene sequence as model,our biosensor exhibits excellent analytical detection limit(50 fM)and high linearity(R2=0.987)over six orders of magnitude dynamic range from 100 fM to 10 nM without any nucleic-acid-amplification assays.The detection can be completed within 1 h with high stability and specificity which benefits from the CRISPR-Cas system.Furthermore,based on the wireless micro-electrochemical platform,the proposed biosensor reveals promising application ability in point-of-care testing.
基金supported by the Nationa l Key Research and Development Program of China (2021YFA1101500)the National Natural Science Foundation of China (22074047)+1 种基金the Hubei Provincial Natural Science Foundation of China (2020CFB578)the Fundamental Research Funds for Central Universities,HUST (2020kfy XJJS034)。
文摘Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.
基金support by the National Key Research and Development Program of China (Grant Nos. 2017FYA0205301, and 2017FYA0205303)the National Natural Science Foundation of China (Grant Nos. 81571835 and 81672247)+3 种基金National Key Research and Development Program of China (No. 2017YFA0205303)National Key Basic Research Program (973 Project) (No. 2015CB931802)"13th Five-Year Plan" Science and Technology Project of Jilin Province Education Department (No. JJKH20170410K)Shanghai Science and Technology Fund (No. 15DZ2252000)
文摘The use of magnetic nanoparticle(MNP)-labeled immunochromatography test strips(ICTSs) is very important for point-ofcare testing(POCT). However, common diagnostic methods cannot accurately analyze the weak magnetic signal from ICTSs, limiting the applications of POCT. In this study, an ultrasensitive multiplex biosensor was designed to overcome the limitations of capturing and normalization of the weak magnetic signal from MNPs on ICTSs. A machine learning model for sandwich assays was constructed and used to classify weakly positive and negative samples, which significantly enhanced the specificity and sensitivity. The potential clinical application was evaluated by detecting 50 human chorionic gonadotropin(HCG) samples and 59 myocardial infarction serum samples. The quantitative range for HCG was 1–1000 mIU mL^(-1) and the ideal detection limit was 0.014 mIU mL^(-1), which was well below the clinical threshold. Quantitative detection results of multiplex cardiac markers showed good linear correlations with standard values. The proposed multiplex assay can be readily adapted for identifying other biomolecules and also be used in other applications such as environmental monitoring, food analysis, and national security.
基金Natural Sciences and Engineering Research Council of Canada (NSERC)Ontario Research Fund (ORF)the India-Canada Centre for Innovative Multidisciplinary Partnerships to Accelerate Community Transformation and Sustainability (IC-IM-PACTS)
文摘The Microchip Imaging Cytometer(MIC)is a class of integrated point-of-care detection systems based on the combination of optical microscopy and flow cytometry.MIC devices have the attributes of portability,cost-effectiveness,and adaptability while providing quantitative measurements to meet the needs of laboratory testing in a variety of healthcare settings.Based on the use of microfluidic chips,MIC requires less sample and can complete sample preparation automatically.Therefore,they can provide quantitative testing results simply using a finger prick specimen.The decreased reagent consumption and reduced form factor also help improve the accessibility and affordability of healthcare services in remote and resource-limited settings.In this article,we review recent developments of the Microchip Imaging Cytometer from the following aspects:clinical applications,microfluidic chip integration,imaging optics,and image acquisition.Following,we provide an outlook of the field and remark on promising technologies that may enable significant progress in the near future.
