Virus is a kind of microorganism and possesses simple structure and contains one nucleic acid,which must be replicated using the host cell system.It causes large-scale infectious diseases and poses serious threats to ...Virus is a kind of microorganism and possesses simple structure and contains one nucleic acid,which must be replicated using the host cell system.It causes large-scale infectious diseases and poses serious threats to the health,social well-being,and economic conditions of millions of people worldwide.Therefore,there is an urgent need to develop novel strategies for accurate diagnosis of virus infection to prevent disease transmission.Quantum dots(QDs)are typical fluorescence nanomaterials with high quantum yield,broad absorbance range,narrow and size-dependent emission,and good stability.QDs-based nanotechnology has been found to be effective method with rapid response,easy operation,high sensitivity,and good specificity,and has been widely applied for the detection of different viruses.However,until now,no systematic and critical review has been published on this important research area.Hence,in this review,we aim to provide a comprehensive coverage of various QDs-based virus detection methods.The fundamental investigations have been reviewed,including information related to the synthesis and biofunctionalization of QDs,QDs-based viral nucleic acid detection strategies,and QDs-based immunoassays.The challenges and perspectives regarding the potential application of QDs for virus detection is also discussed.展开更多
COVID-19 has devastated numerous nations around the world and has overburdened numerous healthcare systems,which has also caused the loss of livelihoods due to prolonged shutdowns and further led to a cascading effect...COVID-19 has devastated numerous nations around the world and has overburdened numerous healthcare systems,which has also caused the loss of livelihoods due to prolonged shutdowns and further led to a cascading effect on the global economy.COVID-19 infections have an incubation period of 2–7 days,but 40 to 45%of cases are asymptomatic or show mild to moderate respiratory symptoms after the period due to subclinical lung abnormalities,making it more likely to spread the pandemic disease.To restrict the spread of the virus,on-site diagnosis methods that are quicker,more precise,and easily accessible are required.Rapid Antigen Detection Tests and Polymerase Chain Reaction tests are currently the primary methods used to determine the presence of COVID-19 viruses.These tests are typically time-consuming,not accurate,and,more importantly,not available to everyone.Hence,in this review and hypothesis,we proposed equipment that employs the properties of photonics to improve the detection of COVID-19 viruses by taking the advantage of typical binding of coronavirus with angiotensin-converting enzyme 2(ACE2)receptors.This hypothetical model would combine Surface-Enhanced Raman Scattering(SERS)and Fluorescence Resonance Energy Transfer(FRET)to provide great flexibility,high sensitivities,and enhanced accessibility.展开更多
Comprehensive Summary:The prevalence and spread of viral infectious diseases pose a grave threat to global public health,particularly resulting in a significant number of casualties.To curb the spread of infectious di...Comprehensive Summary:The prevalence and spread of viral infectious diseases pose a grave threat to global public health,particularly resulting in a significant number of casualties.To curb the spread of infectious diseases,virus testing is one of the efficient and economical means,among which nucleic acid detection has the advantage of detecting viral infections at an early stage,even before symptoms appear.Rolling circle replication is a representative of enzyme-mediated nucleic acid isothermal amplification technology,which is characterized by its mild reaction conditions,no need for temperature control equipment,and high efficiency.This review provides a conceptual overview of the latest advances of rolling circle replication(RCR)and their applications in viral detection,focusing on the molecular design principles in different application scenarios.The first part briefly describes the significance and advantages of RCR in virus detection.The second part elaborates on the design principle and preparation strategy of RCR and its derivative technologies.The third part focuses on the various application scenarios in virus detection.In the end,we provide a perspective on the innovation of RCR in improving the accuracy and specificity of virus detection to cope with the challenges of infectious diseases that may arise in the future.展开更多
Outbreaks of infectious viruses offer a formidable challenge to public healthcare systems and early detection of viruses is essential for preventing virus propagation.In this work,an ultrasensitive plasmon-enhanced fl...Outbreaks of infectious viruses offer a formidable challenge to public healthcare systems and early detection of viruses is essential for preventing virus propagation.In this work,an ultrasensitive plasmon-enhanced fluorescence resonance energy transfer(FRET)biosensor based on core-shell upconversion nanoparticle(csUCNP)and gold nanoparticle(AuNP)for accurate detection of SARS-CoV-2 viral RNA is presented.In this biodetection assay,the Tm^(3+)/Er^(3+)co-doped csUCNP NaGdF_(4):Yb/Tm@NaYF_(4):Yb/Er acts as an energy donor and AuNP serves as an energy acceptor.The upconversion emission of Tm^(3+)and the design of the core-shell structure led to a simultaneous surface plasmon effect of AuNP.The localized surface plasmon resonance(LSPR)arising from collective oscillations of free electrons significantly enhanced FRET efficiency between Er^(3+)and AuNP.The as-prepared biosensor obtained a limit of detection(LOD)as low as 750 aM,indicating that the integration of FRET and surface plasmon into one biodetection assay significantly boosted the sensitivity of the biosensor.In addition,samples extracted from clinical samples are also utilized to validate the effectiveness of the biosensor.Therefore,this innovative plasmon-enhanced FRET biosensor based on Tm^(3+)/Er^(3+)co-doped csUCNP may pave the way for rapid and accurate biodetection applications.展开更多
Rapid and accurate detection of infectious virus particles, not just viral nucleic acid, is essential to avoid unnecessary quarantine and effectively control the spread of viral diseases such as coronavirus disease 20...Rapid and accurate detection of infectious virus particles, not just viral nucleic acid, is essential to avoid unnecessary quarantine and effectively control the spread of viral diseases such as coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome (SARS), and Middle East respiratory syndrome (MERS). Real-time quantitative polymerase chain reaction (RT-qPCR) was the most widely used detection technique during the COVID-19 outbreak. However, it cannot discriminate between intact infectious viruses and surface-distorted, non-infectious virus particles or naked viral RNA. In this study, we present a strategy for the specific detection of infectious coronaviruses by combining viral receptor capture and reverse transcription loop-mediated isothermal amplification (RT-LAMP). We successfully applied this strategy to detect infectious virus particles of the SARS-CoV-2 surrogate virus and the human coronavirus NL63 (HCoV-NL63). Virus particles were first captured on ELISA plates coated with the recombinant human angiotensin-converting enzyme 2 (hACE2) receptor. Viral RNA was then extracted from the particles and detected by RT-LAMP using virus-specific primers. In our experimental setting, the proposed method had a minimum detection limit (LOD) of 90 PFU/mL, sensitivity of 96.2%, and specificity of 100%. Our study provides a proof-of-concept that viral receptor capture combined with RT-LAMP can differentiate infectious coronaviruses from non-infectious virions or naked viral RNA. This paves the way for this virus detection strategy to become a mainstream tool for the management, prevention and control of epidemic coronavirus diseases.展开更多
Cost-effective, rapid, and accurate virus detection technologies play key roles in reducing viral transmission. Prompt and accurate virus detection enables timely treatment and effective quarantine of virus carrier, a...Cost-effective, rapid, and accurate virus detection technologies play key roles in reducing viral transmission. Prompt and accurate virus detection enables timely treatment and effective quarantine of virus carrier, and therefore effectively reduces the possibility of large-scale spread. However, conventional virus detection techniques often suffer from slow response, high cost or sophisticated procedures. Recently, two-dimensional(2D) materials have been used as promising sensing platforms for the highperformance detection of a variety of chemical and biological substances. The unique properties of 2D materials, such as large specific area, active surface interaction with biomolecules and facile surface functionalization, provide advantages in developing novel virus detection technologies with fast response and high sensitivity. Furthermore, 2D materials possess versatile and tunable electronic, electrochemical and optical properties, making them ideal platforms to demonstrate conceptual sensing techniques and explore complex sensing mechanisms in next-generation biosensors. In this review, we first briefly summarize the virus detection techniques with an emphasis on the current efforts in fighting again COVID-19. Then, we introduce the preparation methods and properties of 2D materials utilized in biosensors, including graphene, transition metal dichalcogenides(TMDs) and other 2D materials. Furthermore, we discuss the working principles of various virus detection technologies based on emerging 2D materials, such as field-effect transistor-based virus detection, electrochemical virus detection, optical virus detection and other virus detection techniques. Then, we elaborate on the essential works in 2D material-based high-performance virus detection. Finally, our perspective on the challenges and future research direction in this field is discussed.展开更多
Along with the evolution of computer viruses, the number of file samples that need to be analyzed has constantly increased. An automatic and robust tool is needed to classify the file samples quickly and efficiently. ...Along with the evolution of computer viruses, the number of file samples that need to be analyzed has constantly increased. An automatic and robust tool is needed to classify the file samples quickly and efficiently. Inspired by the human immune system, we developed a local concentration based virus detection method, which connects a certain number of two-element local concentration vectors as a feature vector. In contrast to the existing data mining techniques, the new method does not remember exact file content for virus detection, but uses a non-signature paradigm, such that it can detect some previously unknown viruses and overcome the techniques like obfuscation to bypass signatures. This model first extracts the viral tendency of each fragment and identifies a set of statical structural detectors, and then uses an information-theoretic preprocessing to remove redundancy in the detectors’ set to generate ‘self’ and ‘nonself’ detector libraries. Finally, ‘self’ and ‘nonself’ local concentrations are constructed by using the libraries, to form a vector with an array of two elements of local concentrations for detecting viruses efficiently. Several standard data mining classifiers, including K -nearest neighbor (KNN), radial basis function (RBF) neural networks, and support vector machine (SVM), are leveraged to classify the local concentration vector as the feature of a benign or malicious program and to verify the effectiveness and robustness of this approach. Experimental results show that the proposed approach not only has a much faster speed, but also gives around 98% of accuracy.展开更多
A reverse-transcription loop-mediated isothermal amplification （RT-LAMP） method was established for the detection of wheat streak mosaic virus （WSMV）. Ac-cording to the conservative regions of the genes that encod...A reverse-transcription loop-mediated isothermal amplification （RT-LAMP） method was established for the detection of wheat streak mosaic virus （WSMV）. Ac-cording to the conservative regions of the genes that encode the coat protein of WSMV, 2 pairs of primers were designed. Final y, the 1st pair of primers was select-ed through the specificity test. The sensitivity test showed the sensitivity of RT-LAMP method was 10 times higher than that of RT-PCR. In addition, the amplifica-tion of target gene could be judged visual y from the presence of fluorescence （cal-cein） in the final reaction system. The RT-LAMP method, established in this study, was rapid, easy, specific and sensitive. Moreover, it did not require sophisticated equip-ment. The RT-LAMP was suitable for the rapid detection of WSMV.展开更多
A real-time RT-PCR （RT-qPCR） assay for the detection of Tahyna virus was developed to monitor Tahyna virus infection in field-collected vector mosquito samples. The targets selected for the assay were S segment sequ...A real-time RT-PCR （RT-qPCR） assay for the detection of Tahyna virus was developed to monitor Tahyna virus infection in field-collected vector mosquito samples. The targets selected for the assay were S segment sequences encoding the nucleocapsid protein from the Tahyna virus. Primers and probes were selected in conserved regions by aligning genetic sequences from various Tahyna virus strains available from GenBank. The sensitivity of the RT-qPCR approach was compared to that of a standard plaque assay in BHK cells. RT-qPCR assay can detect 4.8 PFU of titrated Tahyna virus. Assay specificities were determined by testing a battery of arboviruses, including representative strains of Tahyna virus and other arthropod-borne viruses from China. Seven strains of Tahyna virus were confirmed as positive; the other seven species of arboviruses could not be detected by RT-qPCR. Additionally, the assay was used to detect Tahyna viral RNA in pooled mosquito samples. The RT-qPCR assay detected Tahyna virus in a sensitive, specific, and rapid manner; these findings support the use of the assay in viral surveillance.展开更多
Bats carry a variety of viruses, and some of them cause public health problems. Macao, which is famous for its gambling industry, has a complex population structure. The globalization in such an international metropol...Bats carry a variety of viruses, and some of them cause public health problems. Macao, which is famous for its gambling industry, has a complex population structure. The globalization in such an international metropolis has enhanced the chance of disease transmission. Therefore, surveillance of zoonotic viruses is necessary for the early warning of potential emerging infectious diseases.Here, we report the first surveillance of bat viruses in Macao. In this study, we collected 1004 samples involving 10 bat species from 7 sites from April 2015 to May 2016, and examined the presence of viruses using nucleic acid-based methods. Coronaviruses, adenoviruses and paramyxoviruses were detected in these samples, with a high prevalence of coronaviruses. While,none was positive for hepatitis A virus, hepatitis E virus or hantavirus. Co-infections are not common in those bat species, but coronavirus HKU6 and adenovirus can be found commonly occurred in Myotis ricketti.展开更多
Ebola virus disease reemerged in Western Africa in 2014.Chinese Center for Disease Control and Prevention dispatched the first Ebola virus（EBOV）detection team to run newly established Sierra Leone-China Friendship B...Ebola virus disease reemerged in Western Africa in 2014.Chinese Center for Disease Control and Prevention dispatched the first Ebola virus（EBOV）detection team to run newly established Sierra Leone-China Friendship Biological Safety Laboratory.The aims of study were to understand epidemiology,clinical manifestations and survival time of EBOV in patient＇s blood.A total of 913specimens were tested between March 11 and April20, 2015. EBOV positivity occurred in 7.37% of the blood and 0.53% in throat swabs.展开更多
With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluo-rescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyn...With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluo-rescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyngeal carcinoma (NPC) patients and 315 controls (normal and patients with other tumors). MA-IgA antibody was positive in 96.8% of the pretreatment NPC patients with a GMT of 1:36.3. MA-IgA detection by this method was more sensitive than EA-IgA detection by IE. In contrast, patients with tumors other than NPC were negative for MA-IgA antibody. 9.1% of VCA-IgA positive persons were MA-IgA positive with a GMT of less than 1:5. No MA-IgA positive was found in VCA-IgA negatives. The results indicated that this method was relatively specific. In the treatment group, the positive rate and GMT of MA-IgA antibody declined with increase in survival time and the decline was faster than VCA-IgA. When recurrence or distant metastasis developed, similar to VCA-IgA and EA-IgA antibodies, the positive rate and GMT of MA-IgA antibody increased to its pretreatment level. Therefore, MA-IgA detection might be valuable in the early diagnosis and monitor of NPC.展开更多
Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 1...Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).展开更多
[Objective] Sweet potato virus disease had a significant harm to the yield and quality of sweet potato, directly causing the degradation of sweet potato vari- eties and even the harvest failure. Therefore, the detecti...[Objective] Sweet potato virus disease had a significant harm to the yield and quality of sweet potato, directly causing the degradation of sweet potato vari- eties and even the harvest failure. Therefore, the detection and removal of sweet potato virus and the establishment of rapid propagation method of sweet potato is of great significance to ensure the stable inheritance of excellent characters of sweet potato, prevent the spread of sweet potato virus and develop sweet potato industry. [Method] With Xiangshu series varieties of sweet potato, Xiangshu 15 and Xiangshu 19 as the research materials, a virus-free culture program was established for meristem tip apex tissue culture of different cultivars, and a rapid propagation method was developed for virus-free seedlings. [Result｜ On the basis of analysis on seedling emergence rate, the optimal addition scheme of plant hormones in the MS culture medium of Xiangshu 15 was 6-BA 3.0 mg/L ＋ NAA 1.0 mg/L, and the opti- mal plant hormone addition scheme for Xiangshu 19 was 6-BA 2.0 mg/L ＋ NAA 0.67 mg/L Under the developed rapid propagation system, the annual reproductive coefficient was up to 49 152, far higher than that （20 000） in field. IConclusionl Based on the actual production, combined with the meristem tip apex tissue culture, a comprehensive prevention and control measure was put forward, which included virus detection, early warning, removal and virus-free seedlings breeding, tt was of great strategic significance to improve the yield and quality of high-quality sweet potato and ensure the healthy development of sweet potato industry in China.展开更多
A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive an...A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV) strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV-containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries.展开更多
In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBan...In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10-5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.展开更多
基金supported by National Key Research and Development Program of China(2021YFA0910900)the National Natural Science Foundation of China(32222044,22104147)+5 种基金Shenzhen Municipal Science and Technology Innovation Council(RCYX20210609103823046)Youth Innovation Promotion Association CAS(2021359)Natural Science Foundation of Guangdong(2020A1515111130)Guangdong Provincial Key Laboratory of Synthetic Genomics(2019B030301006)Shenzhen Science and Technology Program(KQTD20180413181837372)Shenzhen Outstanding Talents Training Fund.
文摘Virus is a kind of microorganism and possesses simple structure and contains one nucleic acid,which must be replicated using the host cell system.It causes large-scale infectious diseases and poses serious threats to the health,social well-being,and economic conditions of millions of people worldwide.Therefore,there is an urgent need to develop novel strategies for accurate diagnosis of virus infection to prevent disease transmission.Quantum dots(QDs)are typical fluorescence nanomaterials with high quantum yield,broad absorbance range,narrow and size-dependent emission,and good stability.QDs-based nanotechnology has been found to be effective method with rapid response,easy operation,high sensitivity,and good specificity,and has been widely applied for the detection of different viruses.However,until now,no systematic and critical review has been published on this important research area.Hence,in this review,we aim to provide a comprehensive coverage of various QDs-based virus detection methods.The fundamental investigations have been reviewed,including information related to the synthesis and biofunctionalization of QDs,QDs-based viral nucleic acid detection strategies,and QDs-based immunoassays.The challenges and perspectives regarding the potential application of QDs for virus detection is also discussed.
文摘COVID-19 has devastated numerous nations around the world and has overburdened numerous healthcare systems,which has also caused the loss of livelihoods due to prolonged shutdowns and further led to a cascading effect on the global economy.COVID-19 infections have an incubation period of 2–7 days,but 40 to 45%of cases are asymptomatic or show mild to moderate respiratory symptoms after the period due to subclinical lung abnormalities,making it more likely to spread the pandemic disease.To restrict the spread of the virus,on-site diagnosis methods that are quicker,more precise,and easily accessible are required.Rapid Antigen Detection Tests and Polymerase Chain Reaction tests are currently the primary methods used to determine the presence of COVID-19 viruses.These tests are typically time-consuming,not accurate,and,more importantly,not available to everyone.Hence,in this review and hypothesis,we proposed equipment that employs the properties of photonics to improve the detection of COVID-19 viruses by taking the advantage of typical binding of coronavirus with angiotensin-converting enzyme 2(ACE2)receptors.This hypothetical model would combine Surface-Enhanced Raman Scattering(SERS)and Fluorescence Resonance Energy Transfer(FRET)to provide great flexibility,high sensitivities,and enhanced accessibility.
基金supported by the National Key Research and Development Program of China(No.2021YFC2103903)National Natural Science Foundation of China(Nos.22422701,52303174)+1 种基金Beijing Natural Science Foundation-Haidian Original Innovation Joint Fund Project(L232086)Fundamental Research Funds for the Central Universities(buctrc202233).
文摘Comprehensive Summary:The prevalence and spread of viral infectious diseases pose a grave threat to global public health,particularly resulting in a significant number of casualties.To curb the spread of infectious diseases,virus testing is one of the efficient and economical means,among which nucleic acid detection has the advantage of detecting viral infections at an early stage,even before symptoms appear.Rolling circle replication is a representative of enzyme-mediated nucleic acid isothermal amplification technology,which is characterized by its mild reaction conditions,no need for temperature control equipment,and high efficiency.This review provides a conceptual overview of the latest advances of rolling circle replication(RCR)and their applications in viral detection,focusing on the molecular design principles in different application scenarios.The first part briefly describes the significance and advantages of RCR in virus detection.The second part elaborates on the design principle and preparation strategy of RCR and its derivative technologies.The third part focuses on the various application scenarios in virus detection.In the end,we provide a perspective on the innovation of RCR in improving the accuracy and specificity of virus detection to cope with the challenges of infectious diseases that may arise in the future.
基金Research Grants Council of the Hong Kong Special Administrative Region,China,Grant/Award Number:C5110-20GFPolyU Internal Research Fund,Grant/Award Numbers:1-CD4S,1-W21GShenzhen-Hong Kong-Macao Technology Research Programme Fund,Grant/Award Number:SGDX2020110309260000。
文摘Outbreaks of infectious viruses offer a formidable challenge to public healthcare systems and early detection of viruses is essential for preventing virus propagation.In this work,an ultrasensitive plasmon-enhanced fluorescence resonance energy transfer(FRET)biosensor based on core-shell upconversion nanoparticle(csUCNP)and gold nanoparticle(AuNP)for accurate detection of SARS-CoV-2 viral RNA is presented.In this biodetection assay,the Tm^(3+)/Er^(3+)co-doped csUCNP NaGdF_(4):Yb/Tm@NaYF_(4):Yb/Er acts as an energy donor and AuNP serves as an energy acceptor.The upconversion emission of Tm^(3+)and the design of the core-shell structure led to a simultaneous surface plasmon effect of AuNP.The localized surface plasmon resonance(LSPR)arising from collective oscillations of free electrons significantly enhanced FRET efficiency between Er^(3+)and AuNP.The as-prepared biosensor obtained a limit of detection(LOD)as low as 750 aM,indicating that the integration of FRET and surface plasmon into one biodetection assay significantly boosted the sensitivity of the biosensor.In addition,samples extracted from clinical samples are also utilized to validate the effectiveness of the biosensor.Therefore,this innovative plasmon-enhanced FRET biosensor based on Tm^(3+)/Er^(3+)co-doped csUCNP may pave the way for rapid and accurate biodetection applications.
基金supported by the National Natural Science Foundation of China(32470160)the National Key Research and Development Program of China(2021YFC2300100,GZNL2023A01008)+2 种基金the Shenzhen Science and Technology Program(#JSGG20200225150431472,JSGG20210901145403012,and JSGG20220301090005007)the“Pearl River Talent Plan”Innovation and Entrepreneurship Team Project of Guangdong Province(2016LJ06Y540 and 2019ZT08Y464)the Science and Technology Program of Guangdong Province,China(2021B1212040017).
文摘Rapid and accurate detection of infectious virus particles, not just viral nucleic acid, is essential to avoid unnecessary quarantine and effectively control the spread of viral diseases such as coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome (SARS), and Middle East respiratory syndrome (MERS). Real-time quantitative polymerase chain reaction (RT-qPCR) was the most widely used detection technique during the COVID-19 outbreak. However, it cannot discriminate between intact infectious viruses and surface-distorted, non-infectious virus particles or naked viral RNA. In this study, we present a strategy for the specific detection of infectious coronaviruses by combining viral receptor capture and reverse transcription loop-mediated isothermal amplification (RT-LAMP). We successfully applied this strategy to detect infectious virus particles of the SARS-CoV-2 surrogate virus and the human coronavirus NL63 (HCoV-NL63). Virus particles were first captured on ELISA plates coated with the recombinant human angiotensin-converting enzyme 2 (hACE2) receptor. Viral RNA was then extracted from the particles and detected by RT-LAMP using virus-specific primers. In our experimental setting, the proposed method had a minimum detection limit (LOD) of 90 PFU/mL, sensitivity of 96.2%, and specificity of 100%. Our study provides a proof-of-concept that viral receptor capture combined with RT-LAMP can differentiate infectious coronaviruses from non-infectious virions or naked viral RNA. This paves the way for this virus detection strategy to become a mainstream tool for the management, prevention and control of epidemic coronavirus diseases.
基金support from ITC via the Hong Kong Branch of National Precious Metals Material Engineering Research Center (NPMM)the Research Grants Council of Hong Kong (AoE/P-701/ 20)+4 种基金the Start-Up Grant (9380100)the grants from the City University of Hong Kong (9610478, 9680314, 7020013, 1886921)the Science Technology and Innovation Committee of Shenzhen Municipality (JCYJ20200109143412311, SGDX2020110309300301, “Preparation of single atoms on transition metal chalcogenides for electrolytic hydrogen evolution”, City U)funding support from the StartUp Grant (7200656, 9610482)Grant from the City University of Hong Kong (7020013)。
文摘Cost-effective, rapid, and accurate virus detection technologies play key roles in reducing viral transmission. Prompt and accurate virus detection enables timely treatment and effective quarantine of virus carrier, and therefore effectively reduces the possibility of large-scale spread. However, conventional virus detection techniques often suffer from slow response, high cost or sophisticated procedures. Recently, two-dimensional(2D) materials have been used as promising sensing platforms for the highperformance detection of a variety of chemical and biological substances. The unique properties of 2D materials, such as large specific area, active surface interaction with biomolecules and facile surface functionalization, provide advantages in developing novel virus detection technologies with fast response and high sensitivity. Furthermore, 2D materials possess versatile and tunable electronic, electrochemical and optical properties, making them ideal platforms to demonstrate conceptual sensing techniques and explore complex sensing mechanisms in next-generation biosensors. In this review, we first briefly summarize the virus detection techniques with an emphasis on the current efforts in fighting again COVID-19. Then, we introduce the preparation methods and properties of 2D materials utilized in biosensors, including graphene, transition metal dichalcogenides(TMDs) and other 2D materials. Furthermore, we discuss the working principles of various virus detection technologies based on emerging 2D materials, such as field-effect transistor-based virus detection, electrochemical virus detection, optical virus detection and other virus detection techniques. Then, we elaborate on the essential works in 2D material-based high-performance virus detection. Finally, our perspective on the challenges and future research direction in this field is discussed.
基金supported by the National Natural Science Foundation of China (Nos. 60673020 and 60875080)the National High-Tech R & D Program of China (No. 2007AA01Z453)
文摘Along with the evolution of computer viruses, the number of file samples that need to be analyzed has constantly increased. An automatic and robust tool is needed to classify the file samples quickly and efficiently. Inspired by the human immune system, we developed a local concentration based virus detection method, which connects a certain number of two-element local concentration vectors as a feature vector. In contrast to the existing data mining techniques, the new method does not remember exact file content for virus detection, but uses a non-signature paradigm, such that it can detect some previously unknown viruses and overcome the techniques like obfuscation to bypass signatures. This model first extracts the viral tendency of each fragment and identifies a set of statical structural detectors, and then uses an information-theoretic preprocessing to remove redundancy in the detectors’ set to generate ‘self’ and ‘nonself’ detector libraries. Finally, ‘self’ and ‘nonself’ local concentrations are constructed by using the libraries, to form a vector with an array of two elements of local concentrations for detecting viruses efficiently. Several standard data mining classifiers, including K -nearest neighbor (KNN), radial basis function (RBF) neural networks, and support vector machine (SVM), are leveraged to classify the local concentration vector as the feature of a benign or malicious program and to verify the effectiveness and robustness of this approach. Experimental results show that the proposed approach not only has a much faster speed, but also gives around 98% of accuracy.
文摘A reverse-transcription loop-mediated isothermal amplification （RT-LAMP） method was established for the detection of wheat streak mosaic virus （WSMV）. Ac-cording to the conservative regions of the genes that encode the coat protein of WSMV, 2 pairs of primers were designed. Final y, the 1st pair of primers was select-ed through the specificity test. The sensitivity test showed the sensitivity of RT-LAMP method was 10 times higher than that of RT-PCR. In addition, the amplifica-tion of target gene could be judged visual y from the presence of fluorescence （cal-cein） in the final reaction system. The RT-LAMP method, established in this study, was rapid, easy, specific and sensitive. Moreover, it did not require sophisticated equip-ment. The RT-LAMP was suitable for the rapid detection of WSMV.
基金supported by the National Science and Technology Major Project of the Ministry of Science and Technology of China(No.2013ZX10004-101)
文摘A real-time RT-PCR （RT-qPCR） assay for the detection of Tahyna virus was developed to monitor Tahyna virus infection in field-collected vector mosquito samples. The targets selected for the assay were S segment sequences encoding the nucleocapsid protein from the Tahyna virus. Primers and probes were selected in conserved regions by aligning genetic sequences from various Tahyna virus strains available from GenBank. The sensitivity of the RT-qPCR approach was compared to that of a standard plaque assay in BHK cells. RT-qPCR assay can detect 4.8 PFU of titrated Tahyna virus. Assay specificities were determined by testing a battery of arboviruses, including representative strains of Tahyna virus and other arthropod-borne viruses from China. Seven strains of Tahyna virus were confirmed as positive; the other seven species of arboviruses could not be detected by RT-qPCR. Additionally, the assay was used to detect Tahyna viral RNA in pooled mosquito samples. The RT-qPCR assay detected Tahyna virus in a sensitive, specific, and rapid manner; these findings support the use of the assay in viral surveillance.
基金financed by the Environment Construction & Capacity Building of GDAS’Research Platform(2016GDASPT-0215)the Science & Technology Planning Project of Guangdong(2013B050800024 and 2015A020209093)Science & Technology Planning Project of Guangzhou(201707010128)
文摘Bats carry a variety of viruses, and some of them cause public health problems. Macao, which is famous for its gambling industry, has a complex population structure. The globalization in such an international metropolis has enhanced the chance of disease transmission. Therefore, surveillance of zoonotic viruses is necessary for the early warning of potential emerging infectious diseases.Here, we report the first surveillance of bat viruses in Macao. In this study, we collected 1004 samples involving 10 bat species from 7 sites from April 2015 to May 2016, and examined the presence of viruses using nucleic acid-based methods. Coronaviruses, adenoviruses and paramyxoviruses were detected in these samples, with a high prevalence of coronaviruses. While,none was positive for hepatitis A virus, hepatitis E virus or hantavirus. Co-infections are not common in those bat species, but coronavirus HKU6 and adenovirus can be found commonly occurred in Myotis ricketti.
基金supported by a China Mega-Project for Infectious Disease(2011ZX10004-101,2012ZX10004215)Major Program of the National Natural Science Foundation of China(81590763)a SKLID Development Grant(2012SKLID102)
文摘Ebola virus disease reemerged in Western Africa in 2014.Chinese Center for Disease Control and Prevention dispatched the first Ebola virus（EBOV）detection team to run newly established Sierra Leone-China Friendship Biological Safety Laboratory.The aims of study were to understand epidemiology,clinical manifestations and survival time of EBOV in patient＇s blood.A total of 913specimens were tested between March 11 and April20, 2015. EBOV positivity occurred in 7.37% of the blood and 0.53% in throat swabs.
文摘With gene engineering EB virus membrane antigen as the diagnostic antigen, indirect immunofluo-rescence (IF) assay was used to detect IgA antibody against EB virus membrane antigen (MA-IgA) in sera from 202 nasopharyngeal carcinoma (NPC) patients and 315 controls (normal and patients with other tumors). MA-IgA antibody was positive in 96.8% of the pretreatment NPC patients with a GMT of 1:36.3. MA-IgA detection by this method was more sensitive than EA-IgA detection by IE. In contrast, patients with tumors other than NPC were negative for MA-IgA antibody. 9.1% of VCA-IgA positive persons were MA-IgA positive with a GMT of less than 1:5. No MA-IgA positive was found in VCA-IgA negatives. The results indicated that this method was relatively specific. In the treatment group, the positive rate and GMT of MA-IgA antibody declined with increase in survival time and the decline was faster than VCA-IgA. When recurrence or distant metastasis developed, similar to VCA-IgA and EA-IgA antibodies, the positive rate and GMT of MA-IgA antibody increased to its pretreatment level. Therefore, MA-IgA detection might be valuable in the early diagnosis and monitor of NPC.
基金partially supported by the National Institutes of Health(grant no.P20GM103646)the United States Department of Agriculture Animal and Plant Health Inspection Service(agreement 14-7428-1041-CA)
文摘Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).
文摘[Objective] Sweet potato virus disease had a significant harm to the yield and quality of sweet potato, directly causing the degradation of sweet potato vari- eties and even the harvest failure. Therefore, the detection and removal of sweet potato virus and the establishment of rapid propagation method of sweet potato is of great significance to ensure the stable inheritance of excellent characters of sweet potato, prevent the spread of sweet potato virus and develop sweet potato industry. [Method] With Xiangshu series varieties of sweet potato, Xiangshu 15 and Xiangshu 19 as the research materials, a virus-free culture program was established for meristem tip apex tissue culture of different cultivars, and a rapid propagation method was developed for virus-free seedlings. [Result｜ On the basis of analysis on seedling emergence rate, the optimal addition scheme of plant hormones in the MS culture medium of Xiangshu 15 was 6-BA 3.0 mg/L ＋ NAA 1.0 mg/L, and the opti- mal plant hormone addition scheme for Xiangshu 19 was 6-BA 2.0 mg/L ＋ NAA 0.67 mg/L Under the developed rapid propagation system, the annual reproductive coefficient was up to 49 152, far higher than that （20 000） in field. IConclusionl Based on the actual production, combined with the meristem tip apex tissue culture, a comprehensive prevention and control measure was put forward, which included virus detection, early warning, removal and virus-free seedlings breeding, tt was of great strategic significance to improve the yield and quality of high-quality sweet potato and ensure the healthy development of sweet potato industry in China.
文摘A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV) strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV-containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries.
基金Fifteenth National Science and Technology Support Program of China(2007BAD86B04-4)
文摘In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10-5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.
