We theoretically and experimentally study the cavity modes in a gyromagnetic photonic crystal (GPC).Because the permeability of gyromagnetic material relies on the dc magnetic field exerted upon it,the band gap of the...We theoretically and experimentally study the cavity modes in a gyromagnetic photonic crystal (GPC).Because the permeability of gyromagnetic material relies on the dc magnetic field exerted upon it,the band gap of the GPC is supposed to be tunable.Based on this,it is expected that the cavity mode in the GPC is also tunable under the variation of dc magnetic fields.The relation between the frequency of the cavity mode and exerted dc magnetic field is theoretically investigated.Furthermore,we measure the frequency variation of the cavity mode in the GPC and demonstrate the tunable property of GPC cavity.展开更多
Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the ...Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the use of immortalized cell lines in the laboratory to screen nucleic acid aptamers for recognition ability of tumor cells in the diseased body.To address this, primary cells that can be stably passaged were isolated and extracted from spontaneous tumors of genetically engineered pancreatic ductal adenocarcinoma model mice in this study.Next, an automated screening instrument for nucleic acid aptamers developed autonomously by our group was used to perform efficient aptamer screening using a limited number of cells, and the obtained nucleic acid aptamers were affinity verified at the cellular level.Finally, to answer the question of the cell growth environment difference on the recognition ability of nucleic acid aptamers, we verified its targeting ability to tumors in vivo on a nude mice xenograft tumor model, and further used a common antitumor drug doxorubicin combined with nucleic acid aptamers to verify the drug loading ability of this aptamer combined with the targeting therapeutic ability.展开更多
Human-specific insertions play important roles in human phenotypes and diseases.Here we reported a 446-bp insertion(Insert-446)in intron 11 of the TBC1D8B gene,located on chromosome X,and traced its origin to a portio...Human-specific insertions play important roles in human phenotypes and diseases.Here we reported a 446-bp insertion(Insert-446)in intron 11 of the TBC1D8B gene,located on chromosome X,and traced its origin to a portion of intron 6 of the EBF1 gene on chromosome 5.Interestingly,Insert-446 was present in the human Neanderthal and Denisovans genomes,and was fixed in humans after human-chimpanzee divergence.We have demonstrated that Insert-446 acts as an enhancer through binding transcript factors that promotes a higher expression of human TBC1D8B gene as compared with orthologs in macaques.In addition,over-expression TBC1D8B promoted cell proliferation and migration through“a dual finger”catalytic mechanism(Arg538 and Gln573)in the TBC domain in vitro and knockdown of TBC1D8B attenuated tumorigenesis in vivo.Knockout of Insert-446 prevented cell proliferation and migration in cancer and normal cells.Our results reveal that the human-specific Insert-446 promotes cell proliferation and migration by upregulating the expression of TBC1D8B gene.These findings provide a significant insight into the effects of human-specific insertions on evolution.展开更多
Extracellular vesicles(EVs)are cell-derived nanosized vesicles widely recognized for their critical roles in various pathophysiological processes.Molecular analysis of EVs is currently being considered an emerging too...Extracellular vesicles(EVs)are cell-derived nanosized vesicles widely recognized for their critical roles in various pathophysiological processes.Molecular analysis of EVs is currently being considered an emerging tool for diseases diagnosis.However,the small size and heterogeneity of EVs has staggered the EVs research for diseases diagnosis.DNA nanotechnology enables self-assembly of versatile DNA nanostructures and has shown enormous potential in assisting EVs biosensing.In this review,we briefly introduce the recent advances in DNA nanotechnology approaches for EVs detection.The approaches were categorized based on the dimension of DNA nanostructures.We provide critical evaluation of these approaches,and summarize the pros and cons of specific methods.Further,we discuss the challenges and future perspectives in this field.展开更多
Canine parvovirus type 2(CPV-2) infection is the most lethal disease of dogs with higher mortality in puppies worldwide.In today’s world,dogs are an integral part of our communities as well as dogs breeding and reari...Canine parvovirus type 2(CPV-2) infection is the most lethal disease of dogs with higher mortality in puppies worldwide.In today’s world,dogs are an integral part of our communities as well as dogs breeding and rearing has become a lucrative business.Therefore,a fast,accurate,portable,and costeffective CPV-2 detection method with the ability for on-site detection is highly desired.In this study,we for the first time proposed a nanosystem for CPV-2 DNA detection with RNA-guided RNA endonuclease Cas13 a,which upon activation results in collateral RNA degradation.We expressed LwCasl3 a in prokaryotic expression system and purified it through nickel column.Activity of Cas13 a was verified by RNA-bound fluorescent group while using a quenched fluorescent probe as signals.Further Cas13 a was combined with Recombinase polymerase amplification(RPA) and T7 transcription to establish molecular detection system termed specific high-sensitivity enzymatic reporter un-locking(SHERLOCK) for sensitive detection of CPV-2 DNA.This nanosystem can detect 100 amol/L CPV-2 DNA within 30 min.The proposed nanosystem exhibited high specificity when tested for CPV-2 and other dog viruses.This CRISPR-Cas13 a mediated sensitive detection approach can be of formidable advantage during CPV-2 outbreaks because it is time-efficient,less laborious and does not involve the use of sophisticated instruments.展开更多
Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electroc...Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electrochemical biosensor is composed of a aptamer as the biological recognition element and transducer converting the biologic interaction into electrical signals for the quantitative measurement of targets.Improvement of the sensitivity of a biosensor is significantly important in order to achieve the detection of biomolecules with low abundance,and different amplification strategies have been explored.The strategies either employ nanomaterials such as gold nanoparticles to con struct electrodes which can transfer the biological reactions more efficiently,or attempt to obtain enhanced signal through multi-labeled carriers or utilize enzyme mimics to catalyze redox cycling.This review discusses recent advances in signal amplification methods and their applications.Critical assessment of each method is also considered.展开更多
Listeriosis is caused by Listeria monocytogenes(LM) and is currently considered to be one of the leading food-borne diseases worldwide, with mortality rate of 20%~30%. Currently, detection methods for LM are time-cons...Listeriosis is caused by Listeria monocytogenes(LM) and is currently considered to be one of the leading food-borne diseases worldwide, with mortality rate of 20%~30%. Currently, detection methods for LM are time-consuming with low sensitivity, and delayed detection results. SYTO9 has a high affinity for DNA and exhibits enhanced fluorescence upon binding. Therefore, this study used SYTO9 staining and image processing to develop a rapid loop mediated isothermal amplification(LAMP) detection method for LM. Smartphone was successfully used for detecting the color change in different concentrations of LM. Besides, the optimized LAMP reaction temperature was 63 °C by color identification, and the limit of detection for LM was 6 copies/μL in the green channel. So, the developed method, based on image processing, is simple, sensitive and rapid, which provides a new idea and method for rapid detection of LM and other food-borne bacterial pathogens.展开更多
Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to dis...Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to discover some novel tumor markers,i.e.,membrane proteins that are specifically highly expressed on the surface of tumor cells.Tumor markers can be used in many fields such as early diagnosis and treatment,and a new type of tumor marker proved to be effective can significantly improve the therapeutic effect of such tumors.However,further characterization of newly acquired membrane proteins is essential for their clinical use as tumor markers.This review first briefly introduced the process of obtaining novel tumor markers from nucleic acid aptamers.Next,the commonly used protein characterization methods could be used as a technical means to identify membrane protein targets corresponding to tumor cell aptamers,to clarify the principles,advantages and disadvantages of various means,and to analyze the most suitable situations for various experimental methods.Finally,the outlook was made and the characterization methods that should be used in such experiments were summarized.展开更多
Point-of-care nucleic acid testing(POCNAT) has played an important role in the outbreak of infectious diseases(e.g., COVID-19) over recent years. POCNAT aims to realize the rapid, simple and automatic detection of nuc...Point-of-care nucleic acid testing(POCNAT) has played an important role in the outbreak of infectious diseases(e.g., COVID-19) over recent years. POCNAT aims to realize the rapid, simple and automatic detection of nucleic acid. Thanks to the development of manufacturing technology, electronic information technology, artificial intelligence technology, and biological information technology in recent years, the development of the POCNAT device has led to significant advancement. Instead of the normal nucleic acid detection methods used in the laboratory, some novel experimental carriers have been applied, such as chips, cartridges and papers. The application of these experimental carriers has realized the automation and integration of nucleic acid detection. The entire process of nucleic acid detection is normally divided into three steps(nucleic acid extraction, target amplification and signal detection). All of the reagents required by the process can be pre-stored on these experimental carriers, without unnecessary manual operation. Furthermore, all of the processes are carried out in this experimental carrier, with the assistance of a specific control device. Although they are complicated to manufacture and precise in design,their application provides a significant step forwards in nucleic acid detection and realizes the integration of nucleic acid detection. This technology has great potential in the field of point-of-care molecular diagnostics in the future. This paper focuses on the relevant content of these experimental carriers.展开更多
Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sen...Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.展开更多
Pathological detection using immunohistochemistry(IHC)has become an indispensable process in the diagnosis confirmation of various cancers.However,the production of monoclonal antibodies is always very complex,expensi...Pathological detection using immunohistochemistry(IHC)has become an indispensable process in the diagnosis confirmation of various cancers.However,the production of monoclonal antibodies is always very complex,expensive and time-consuming,and the batch differences are significant due to the corporeity and health statuses of animals may be different.In this work,an aptamer-based histochemistry(aptahistochemistry)assay was developed using a DNA aptamer for specific diagnosis of clinical breast cancer tissue sections.This aptahistochemistry assay can specifically distinguish Luminal A breast cancer molecular subtype from Luminal B(HER2+),HER2-enriched,and triple-negative breast cancer molecular subtypes,as well as para-carci noma tissue,mastitis tissue and normal breast tissue.The accuracy of this aptahistochemistry assay for the diagnosis of Luminal A breast cancer was as high as 80%,which showed a great potential for clinical pathological diagnosis applications.展开更多
This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the ...This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional strip.The results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic strip.With the results,the determination correlation of the developed microfluidic chip has been greatly improved.In addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional strip.The biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.展开更多
A simple visual method for DNA detection during the formation of gold nanoparticles(AuNPs) was developed based on different electrostatic properties of single strand DNA(ssDNA) and double strand DNA(dsDNA).Since the s...A simple visual method for DNA detection during the formation of gold nanoparticles(AuNPs) was developed based on different electrostatic properties of single strand DNA(ssDNA) and double strand DNA(dsDNA).Since the ssDNA is easy to bind to AuNPs due to its exposed bases which could prevent saltinduced aggregation of AuNPs.The dsDNA always present negative charge because its negatively charged phosphate backbone is exposed.In this case,the dsDNA could disturb the adsorption between dsDNA and AuNPs and result in non-aggregation of AuNPs.After hybridization,chloroauric acid and ascorbic acid were added to the mixture solution,and the solution changed to red immediately and turned to purple in10 min in the present of target DNA.TEM results confirmed that the change of color stemed from aggregation of AuNPs.In order to obtain accurate results by naked eye,the DNA detection assay should be conducted under pH 7.0.展开更多
基金Supported by the National Natural Science Foundation of China under Grant No 10904167the National Basic Research Program of China under Grant No 2011CB922002.
文摘We theoretically and experimentally study the cavity modes in a gyromagnetic photonic crystal (GPC).Because the permeability of gyromagnetic material relies on the dc magnetic field exerted upon it,the band gap of the GPC is supposed to be tunable.Based on this,it is expected that the cavity mode in the GPC is also tunable under the variation of dc magnetic fields.The relation between the frequency of the cavity mode and exerted dc magnetic field is theoretically investigated.Furthermore,we measure the frequency variation of the cavity mode in the GPC and demonstrate the tunable property of GPC cavity.
基金supported by the National Key Research and Development Program of China(Nos.2017YFA0205301 and 2018YFC1602905)National Natural Science Foundation of China(Nos.62071119,62075098,81902153,61527806)the Open Funding of State Key Laboratory of Oral Diseases(No.SKLOD2022OF05).
文摘Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the use of immortalized cell lines in the laboratory to screen nucleic acid aptamers for recognition ability of tumor cells in the diseased body.To address this, primary cells that can be stably passaged were isolated and extracted from spontaneous tumors of genetically engineered pancreatic ductal adenocarcinoma model mice in this study.Next, an automated screening instrument for nucleic acid aptamers developed autonomously by our group was used to perform efficient aptamer screening using a limited number of cells, and the obtained nucleic acid aptamers were affinity verified at the cellular level.Finally, to answer the question of the cell growth environment difference on the recognition ability of nucleic acid aptamers, we verified its targeting ability to tumors in vivo on a nude mice xenograft tumor model, and further used a common antitumor drug doxorubicin combined with nucleic acid aptamers to verify the drug loading ability of this aptamer combined with the targeting therapeutic ability.
基金supported by Key Research and Development Program of Yunnan(202203AC100010)the National Natural Science Foundation of China(31760311,32160236,81830087,U2102203)+4 种基金the National Key Research and Development Program of China(2022YFC2601604,2018YFC2000400,2020YFA0112300)Spring City Plan:the Highlevel Talent Promotion and Training Project of Kunming(2022SCP001)the Yunnan Fundamental Research Projects(CY22624104,202101AS070050)the open project of State Key Laboratory of Genetic Resources and Evolution,Kunming Institute of Zoology,Chinese Academy of Sciences(GREKF17-01)Yunnan University's new round of"Double First-Class"Construction Project—For People’s Life and Health(CY22624104)。
文摘Human-specific insertions play important roles in human phenotypes and diseases.Here we reported a 446-bp insertion(Insert-446)in intron 11 of the TBC1D8B gene,located on chromosome X,and traced its origin to a portion of intron 6 of the EBF1 gene on chromosome 5.Interestingly,Insert-446 was present in the human Neanderthal and Denisovans genomes,and was fixed in humans after human-chimpanzee divergence.We have demonstrated that Insert-446 acts as an enhancer through binding transcript factors that promotes a higher expression of human TBC1D8B gene as compared with orthologs in macaques.In addition,over-expression TBC1D8B promoted cell proliferation and migration through“a dual finger”catalytic mechanism(Arg538 and Gln573)in the TBC domain in vitro and knockdown of TBC1D8B attenuated tumorigenesis in vivo.Knockout of Insert-446 prevented cell proliferation and migration in cancer and normal cells.Our results reveal that the human-specific Insert-446 promotes cell proliferation and migration by upregulating the expression of TBC1D8B gene.These findings provide a significant insight into the effects of human-specific insertions on evolution.
基金supported by the National Natural Science Foundation of China(Nos.82002242,81902153 and 62071119)Natural Science Foundation of Jiangsu Province(No.BK20200135)+3 种基金Hunan Key R&D Projects(No.2021SK2003)Key Project supported by Medical Science and Technology Development Foundation,Nanjing Department of Health(No.YKK20054)Nanjing Important Science&Technology Specific Projects(No.2021-11005)open Funding of State Key Laboratory of Oral Diseases(No.SKLOD2022OF05)。
文摘Extracellular vesicles(EVs)are cell-derived nanosized vesicles widely recognized for their critical roles in various pathophysiological processes.Molecular analysis of EVs is currently being considered an emerging tool for diseases diagnosis.However,the small size and heterogeneity of EVs has staggered the EVs research for diseases diagnosis.DNA nanotechnology enables self-assembly of versatile DNA nanostructures and has shown enormous potential in assisting EVs biosensing.In this review,we briefly introduce the recent advances in DNA nanotechnology approaches for EVs detection.The approaches were categorized based on the dimension of DNA nanostructures.We provide critical evaluation of these approaches,and summarize the pros and cons of specific methods.Further,we discuss the challenges and future perspectives in this field.
基金supported by the National Key Research and Development Program of China (No.2017YFA0205301)National Natural Science Foundation of China (Nos.81902153,61527806 and 81430055)+2 种基金Key Research and Development Project of Jiangsu Province (No.BE2019761)Programs for Changjiang Scholars and Innovative Research Team in University (No.IRT_15R13)open Funding of State Key Laboratory of Oral Diseases (No.SKLOD20190F03)
文摘Canine parvovirus type 2(CPV-2) infection is the most lethal disease of dogs with higher mortality in puppies worldwide.In today’s world,dogs are an integral part of our communities as well as dogs breeding and rearing has become a lucrative business.Therefore,a fast,accurate,portable,and costeffective CPV-2 detection method with the ability for on-site detection is highly desired.In this study,we for the first time proposed a nanosystem for CPV-2 DNA detection with RNA-guided RNA endonuclease Cas13 a,which upon activation results in collateral RNA degradation.We expressed LwCasl3 a in prokaryotic expression system and purified it through nickel column.Activity of Cas13 a was verified by RNA-bound fluorescent group while using a quenched fluorescent probe as signals.Further Cas13 a was combined with Recombinase polymerase amplification(RPA) and T7 transcription to establish molecular detection system termed specific high-sensitivity enzymatic reporter un-locking(SHERLOCK) for sensitive detection of CPV-2 DNA.This nanosystem can detect 100 amol/L CPV-2 DNA within 30 min.The proposed nanosystem exhibited high specificity when tested for CPV-2 and other dog viruses.This CRISPR-Cas13 a mediated sensitive detection approach can be of formidable advantage during CPV-2 outbreaks because it is time-efficient,less laborious and does not involve the use of sophisticated instruments.
基金This work was financially supported by the National Key Research and Development Program of China(No.2017YFA0205301)the National Natural Science Foundation of China(Nos.61527806,81902153 and 61871180)+1 种基金the Clinical Advanced Technology of Social Development Projects in Jiangsu Province(No.BE2018695)the Natural Science Foundation of Hunan Province(No.2017JJ2069).
文摘Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electrochemical biosensor is composed of a aptamer as the biological recognition element and transducer converting the biologic interaction into electrical signals for the quantitative measurement of targets.Improvement of the sensitivity of a biosensor is significantly important in order to achieve the detection of biomolecules with low abundance,and different amplification strategies have been explored.The strategies either employ nanomaterials such as gold nanoparticles to con struct electrodes which can transfer the biological reactions more efficiently,or attempt to obtain enhanced signal through multi-labeled carriers or utilize enzyme mimics to catalyze redox cycling.This review discusses recent advances in signal amplification methods and their applications.Critical assessment of each method is also considered.
基金supported by the grant from the National Natural Science Foundation of China (Nos. 61901168, 8200240581902153)+1 种基金Zhuzhou Innovative City Construction Project(No. 2020–020)China Postdoctoral Science Foundation (No.2018M630498)。
文摘Listeriosis is caused by Listeria monocytogenes(LM) and is currently considered to be one of the leading food-borne diseases worldwide, with mortality rate of 20%~30%. Currently, detection methods for LM are time-consuming with low sensitivity, and delayed detection results. SYTO9 has a high affinity for DNA and exhibits enhanced fluorescence upon binding. Therefore, this study used SYTO9 staining and image processing to develop a rapid loop mediated isothermal amplification(LAMP) detection method for LM. Smartphone was successfully used for detecting the color change in different concentrations of LM. Besides, the optimized LAMP reaction temperature was 63 °C by color identification, and the limit of detection for LM was 6 copies/μL in the green channel. So, the developed method, based on image processing, is simple, sensitive and rapid, which provides a new idea and method for rapid detection of LM and other food-borne bacterial pathogens.
基金financially supported by the National Key Researchand Development Program of China(No.2018YFC1602905)National Natural Science Foundation of China(Nos.81902153,61871180,61527806)。
文摘Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to discover some novel tumor markers,i.e.,membrane proteins that are specifically highly expressed on the surface of tumor cells.Tumor markers can be used in many fields such as early diagnosis and treatment,and a new type of tumor marker proved to be effective can significantly improve the therapeutic effect of such tumors.However,further characterization of newly acquired membrane proteins is essential for their clinical use as tumor markers.This review first briefly introduced the process of obtaining novel tumor markers from nucleic acid aptamers.Next,the commonly used protein characterization methods could be used as a technical means to identify membrane protein targets corresponding to tumor cell aptamers,to clarify the principles,advantages and disadvantages of various means,and to analyze the most suitable situations for various experimental methods.Finally,the outlook was made and the characterization methods that should be used in such experiments were summarized.
基金supported by the National Natural Science Foundation of China (Nos. 61901168, 61971187, 61871180, 61571187, 81902153)Zhuzhou Innovative City Construction Project (No. 2020-020)+2 种基金China Postdoctoral Science Foundation (No. 2018M630498)Hunan Urgency Project (No. 2020SK3005)Education Department Outstanding Young Project of Hunan Province (No. 18B299)。
文摘Point-of-care nucleic acid testing(POCNAT) has played an important role in the outbreak of infectious diseases(e.g., COVID-19) over recent years. POCNAT aims to realize the rapid, simple and automatic detection of nucleic acid. Thanks to the development of manufacturing technology, electronic information technology, artificial intelligence technology, and biological information technology in recent years, the development of the POCNAT device has led to significant advancement. Instead of the normal nucleic acid detection methods used in the laboratory, some novel experimental carriers have been applied, such as chips, cartridges and papers. The application of these experimental carriers has realized the automation and integration of nucleic acid detection. The entire process of nucleic acid detection is normally divided into three steps(nucleic acid extraction, target amplification and signal detection). All of the reagents required by the process can be pre-stored on these experimental carriers, without unnecessary manual operation. Furthermore, all of the processes are carried out in this experimental carrier, with the assistance of a specific control device. Although they are complicated to manufacture and precise in design,their application provides a significant step forwards in nucleic acid detection and realizes the integration of nucleic acid detection. This technology has great potential in the field of point-of-care molecular diagnostics in the future. This paper focuses on the relevant content of these experimental carriers.
基金financially supported by National Natural Science Foundation of China (Nos.81902153,61871180,62071119 and 61971187)Jiangsu Provincial Key Research and Development Program (Nos.BA2020016 and BE 2018695)。
文摘Chemiluminescence immunoassay(CLⅠA) has always been a great challenge in detecting cardiac troponin Ⅰ(c Tn Ⅰ) in whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Ⅰn this study, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotinconjugated c Tn Ⅰ antibody and detected by streptavidin/acridine aster-conjugated polychloromethylstyrene microspheres(PCMS). After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the c Tn Ⅰ concentrations of the serum samples,plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62%~5.67%. The assay was linear over the studied range of 0.01-50.00 ng/mL, and no hook effect was found when c Tn Ⅰ concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit(Abbott assay kit), the correlation coefficient was 0.9859. A washing-free CLⅠA was established for the rapid detection of c Tn Ⅰ in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and PCMS for signal amplification, which showed great potential in clinical application.
基金supported by the National Key Research and Development Program of China (No.2017YFA0205301)the National Natural Science Foundation of China (Nos.61527806, 81902153 and 81771976)the Scientific Research Foundation of the Graduate School of Southeast University (No.YBPY1881)
文摘Pathological detection using immunohistochemistry(IHC)has become an indispensable process in the diagnosis confirmation of various cancers.However,the production of monoclonal antibodies is always very complex,expensive and time-consuming,and the batch differences are significant due to the corporeity and health statuses of animals may be different.In this work,an aptamer-based histochemistry(aptahistochemistry)assay was developed using a DNA aptamer for specific diagnosis of clinical breast cancer tissue sections.This aptahistochemistry assay can specifically distinguish Luminal A breast cancer molecular subtype from Luminal B(HER2+),HER2-enriched,and triple-negative breast cancer molecular subtypes,as well as para-carci noma tissue,mastitis tissue and normal breast tissue.The accuracy of this aptahistochemistry assay for the diagnosis of Luminal A breast cancer was as high as 80%,which showed a great potential for clinical pathological diagnosis applications.
基金financially supported by National Natural Science Foundation of China(Nos.81902153,61871180,62071119 and 61971187)Jiangsu Provincial Key Research and Development Program(No.BE 2018695)。
文摘This device is aimed at ensuring that the sample is uniformly and equivalently reacted with the antibody on the NC membrane in each test when the microfluidic liquid system is introduced to the chip.In this study,the developed microfluidic chip can avoid the presence of the sample and conjugate pads in the chip,while the precision of the chro matography system can be greatly improved using the same particles,NC membrane and antibody alongside the traditional strip.The results,taking the detection of cTnI as an example,revealed that the coefficient of variation(CV)is controlled within 4%,while the maximum record of the contrast chromatographic reagent strip can reach 15%.Additionally,the detection sensitivity can maintain the same order of magnitudes with that of the traditional chromatographic strip.With the results,the determination correlation of the developed microfluidic chip has been greatly improved.In addition,the CV of the chip in this study is greatly improved in comparison with that of the traditional strip.The biggest improvement lies in the mixing between the sample and the microspheres,indicating that this is a new approach to improve the CV of the traditional strip.
基金financially supported by the National Key Research and Development Program of China (No.2017YFA0205301)the National Natural Science Foundation of China (Nos.61527806, 81902153 and 61871180)+2 种基金the Natural Science Foundation of Hunan Province (No.2017JJ2069)the Hunan Key Research Project (No.2017SK2174)the Programs for Changjiang Scholars and Innovative Research Team in University (No.IRT_15R13)
文摘A simple visual method for DNA detection during the formation of gold nanoparticles(AuNPs) was developed based on different electrostatic properties of single strand DNA(ssDNA) and double strand DNA(dsDNA).Since the ssDNA is easy to bind to AuNPs due to its exposed bases which could prevent saltinduced aggregation of AuNPs.The dsDNA always present negative charge because its negatively charged phosphate backbone is exposed.In this case,the dsDNA could disturb the adsorption between dsDNA and AuNPs and result in non-aggregation of AuNPs.After hybridization,chloroauric acid and ascorbic acid were added to the mixture solution,and the solution changed to red immediately and turned to purple in10 min in the present of target DNA.TEM results confirmed that the change of color stemed from aggregation of AuNPs.In order to obtain accurate results by naked eye,the DNA detection assay should be conducted under pH 7.0.
