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Development of an Integrated Disposable Device for SARSCoV-2 nucleic acid Extraction and detection

Biomedical and Environmental Sciences 2024年第6期37卷 639-646页

作者： Jing Ma Yanzhe Hao Meiling Hou Xiaoshan Zhang Jingduan Liu Haodi Meng Jiangbo Chang Xuejun Ma Jihua Liu Qingjie Ying Xianhua Wang Hongxia Li Yuxi Cao Xiaoguang Zhang National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Beijing Runbio Biotechnology Development Co. LtdBeijing 100106China Beijing Beta Technology Co. LtdBeijing 101318China Jiangsu Qitian Gene Biotechnology Co. LtdWuxi 214135JiangsuChina

Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Methods We designed,developed,and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and *** precision of the liquid transfer and temperature control was tested.A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process,from SARS-CoV-2 nucleic acid extraction to amplification,was *** The precision of the syringe transfer volume was 19.2±1.9μL(set value was 20),32.2±1.6(set value was 30),and 57.2±3.5(set value was 60).Temperature control in the amplification tube was measured at 60.0±0.0℃(set value was 60)and 95.1±0.2℃(set value was 95)***-Cov-2 nucleic acid extraction yield through the device was 7.10×10^(6) copies/mL,while a commercial kit yielded 2.98×10^(6) copies/*** mean time to complete the entire assay,from SARS-CoV-2 nucleic acid extraction to amplification detection,was 36 min and 45 *** detection limit for SARS-CoV-2 nucleic acid was 250 copies/*** The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).

关键词： An integrated disposable device SARS-Cov-2 nucleic acid detection

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nucleic acid detection in the diagnosis and prevention of schistosomiasis

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Infectious Diseases of Poverty 2016年第1期5卷 230-240页

作者： Ping He Lan-gui Song Hui Xie Jin-yi Liang Dong-ya Yuan Zhong-dao Wu Zhi-yue Lv Department of Parasitology Zhongshan School of MedicineSun Yat-sen UniversityGuangzhou 510080China Key Laboratory of Tropical Disease Control Ministry of EducationGuangzhou 510080China Department of Pathogenic Biology Medical SchoolXizang Minzu UniversityXianyang 712082China

Schistosomiasis is an important zoonotic parasitic disease that causes serious harms to humans and *** and diagnosis play key roles in schistosomiasis control,however,current techniques for surveillance and diagnosis of the disease have *** genome data for parasites are increasing,novel techniques for detection incorporating nucleotide sequences are receiving widespread *** sensitive,specific,and rapid detection methods are particularly important in the diagnosis of low-grade and early infections,and may prove to have clinical *** paper reviews the progress of nucleic acid detection in the diagnosis and prevention of schistosomiasis,including such aspects as the selection of target genes,and development and application of nucleic acid detection methods.

关键词： Schistosoma japonicum S.mansoni S.haematobium Diagnosis nucleic acid detection

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An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system

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Science China Chemistry 2022年第3期65卷 630-640页

作者： Zhen Sun Kang-Feng Lin Ze-Hang Zhao Yang Wang Xin-Xin Hong Jian-Guang Guo Qing-Yu Ruan Lian-Yu Lu Xiao Li Rui Zhang Chao-Yong Yang Bo-An Li State Key Laboratory of Cellular Stress Biology Innovation Center for Cell Signaling Network and Engineering Research Center of Molecular Diagnostics of the Ministry of EducationSchool of Life SciencesXiamen UniversityXiamen 361005China MOE Key Laboratory of Spectrochemical Analysis&Instrumentation Key Laboratory of Chemical Biology of Fujian ProvinceState Key Laboratory of Physical Chemistry of Solid SurfacesCollaborative Innovation Center of Chemistry for Energy MaterialsDepartment of Chemical EngineeringDepartment of Chemical BiologyCollege of Chemistry and Chemical EngineeringXiamen UniversityXiamen 361005China Department of Thoracic Surgery The First Affiliated HospitalXiamen UniversityXiamen 361005China

Outbreaks of both influenza virus and the novel coronavirus SARS-CoV-2 are serious threats to human health and life. It is very important to establish a rapid, accurate test with large-scale detection potential to prevent the further spread of the epidemic. An optimized RPA-Cas12a-based platform combined with digital microfluidics(DMF), the RCD platform, was established to achieve the automated, rapid detection of influenza viruses and SARS-CoV-2. The probe in the RPA-Cas12a system was optimized to produce maximal fluorescence to increase the amplification signal. The reaction droplets in the platform were all at the microliter level and the detection could be accomplished within 30 min due to the effective mixing of droplets by digital microfluidic technology. The whole process from amplification to recognition is completed in the chip, which reduces the risk of aerosol contamination. One chip can contain multiple detection reaction areas, offering the potential for customized *** RCD platform demonstrated a high level of sensitivity, specificity(no false positives or negatives), speed(≤30 min),automation and multiplexing. We also used the RCD platform to detect nucleic acids from influenza patients and COVID-19 patients. The results were consistent with the findings of q PCR. The RCD platform is a one-step, rapid, highly sensitive and specific method with the advantages of digital microfluidic technology, which circumvents the shortcomings of manual operation. The development of the RCD platform provides potential for the isothermal automatic detection of nucleic acids during epidemics.

关键词： nucleic acid detection digital microfluidics Cas12a influenza virus SARS-CoV-2

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A rapid nucleic acid detection platform based on phosphorothioate-DNA and sulfur binding domain

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Synthetic and Systems Biotechnology 2023年第2期8卷 213-219页

作者： Yuting Shuai Yi Ju Yuanhang Li Dini Ma Lan Jiang Jingyu Zhang Gao-Yi Tan Xueting Liu Shenlin Wang Lixin Zhang Guang Liu State Key Laboratory of Bioreactor Engineering and School of BiotechnologyEast China University of Science and Technology(ECUST)Shanghai200237China State Key Laboratory of Microbial Metabolism Joint International Research Laboratory of Metabolic&Developmental SciencesSchool of Life Sciences&BiotechnologyShanghai Jiao Tong UniversityShanghai200240China

nucleic acid detection plays a key role in diverse diagnosis and disease *** available nucleic acid detection techniques are challenged by trade-offs among speed,simplicity,precision and ***,we described a novel method,designated SENSOR(Sulfur DNA mediated nucleic acid sensing platform),for rapid nucleic acid *** was developed from phosphorothioate(PT)-DNA and sulfur binding domain(SBD)which specifically binds double-stranded PT-modified *** utilizes PT-DNA oligo and SBD as targeting module,which is linked with split luciferase reporter to generate luminescence signal within 10 *** tested detection on synthesized nucleic acid and COVID-19 pseudovirus,achieving attomolar sensitivity combined with an amplification *** nucleotide polymorphisms(SNP)could also be *** SENSOR a new promising nucleic acid detection technique.

关键词： Phosphorothioate Sulfur binding domain nucleic acid detection

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Recent developments and trends of automatic nucleic acid detection systems

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Journal of Biosafety and Biosecurity 2022年第1期4卷 54-58页

作者： Xujun Yuan Guodong Sui Dawei Zhang Min Chen Wang Zhao Shanghai Environmental Biosafety Instruments and Equipment Engineering Technology Research Center University of Shanghai for Science and Technology516 Jungong RoadShanghai 200093PR China Shanghai Cohere Electronic Technology Company No.10 Building2888 Shengang RoadShanghai 201101PR China Shanghai Key Laboratory of Atmospheric Particle Pollution Prevention(LAP3) Department of Environmental Science&EngineeringFudan University2005 Songhu RoadShanghai 200438PR China Engineering Research Center of Optical Instrument and System the Ministry of EducationShanghai Key Laboratory of Modern Optical SystemUniversity of Shanghai for Science and Technology516 Jungong RoadShanghai 200093PR China Shanghai Municipal Center for Disease Control&Prevention 1380 West Zhongshan RoadShanghai 200336PR China

nucleic acid detection,widely used in clinical diagnosis,biological analysis,and environmental monitoring,is of great significance for disease diagnosis and basic *** the outbreak of COVID-19,the demand for fast and high-throughput nucleic acid detection from large numbers of samples has increased *** nucleic acid detection systems can meet these needs,and also play important roles in disease screening and infectious disease prevention and *** this review,we introduce and compare the current mainstream nucleic acid automatic detection instruments and equipment,then discuss the future demands of nucleic acid detection.

关键词： nucleic acid detection Automatic systems

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Ultrathin metal-organic framework nanosheets (Cu-TCPP)-based isothermal nucleic acid amplification for food allergen detection

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Food Science and Human Wellness 2023年第5期12卷 1788-1798页

作者： Jiale Gao Xiaodong Sun Yongxin Liu Bing Niu Qin Chen Xueen Fang Shanghai Key Laboratory of Bio-Energy Crops School of Life SciencesShanghai UniversityShanghai 200444China Shanghai Engineering Research Center of Organ Repair School of MedicineShanghai UniversityShanghai 200444China Department of Chemistry and Institutes of Biomedical Sciences Fudan UniversityShanghai 200433China

The rapid and accurate detection of peanuts and soybeans allergen is important to the food safety. In this study, Cu-TCPP nanosheet, a kind of ultra-thin metal-organic framework(MOF)was synthesized and applied in loop-mediated isothermal amplification(named Cu-TCPP@LAMP), which can inhibit the non-specific amplification by absorbing and precise temperature releasing of single primer. As thus, Cu-TCPP@LAMP can achieve high sensitivity and specific amplification of the target gene. As a result, peanut and soybean allergens genes contained in food were successfully detected with a favorable detection sensitivity(5 ng/μL for peanuts and 10 ng/μL for soybeans)and reliable repeatability(The coefficient of variation was 3.38% for peanuts and 3.33% for soybeans). Moreover, the established method was utilized for detection of several commercial products, and had a high consistency with the standard method. Apart from food allergens, this novel assay can be widely used in other areas, such as pathogen detection, tumor nucleic acid detection and so on.

关键词： Cu-TCPP nanosheet Plant allergen nucleic acid detection Loop-mediated isothermal amplification

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A novel cartridge for nucleic acid extraction,amplification and detection of infectious disease pathogens with the help of magnetic nanoparticles

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中国化学快报:英文版 2023年第8期34卷 236-240页

作者： Yile Fang Yue Wang Liangxi Zhu Haoran Liu Xiangyi Su Yuan Liu Zhu Chen Hui Chen Nongyue He State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province Hunan Key Laboratory of Green Chemistry and Application of Biological NanotechnologyHunan University of TechnologyZhuzhou 412007China

nucleic acid detection(NAD)based on real-time polymerase chain reaction(real-time PCR)is gold standard for infectious disease *** nanoparticles(MNPs)are widely used for nucleic acid extraction(NAE)because of their excellent *** technology makes automated NAD ***,most of the NAD microfluidic chips are too complex to be applied to point-of-care(POC)*** this paper,a simple-structure cartridge was developed for POC detection of infectious *** self-contained cartridge can be divided into a magnetic-controlled NAE part,a valve-piston combined fluidic control part and a PCR chip,which is able to extract nucleic acid from up to 500μL of liquid samples by MNPs and finish the detection process from“sample in”to“answer out”*** tests of the cartridges show that it met the demands of automated *** of on-cartridge detection of hepatitis B virus(HBV)demonstrated that this system has good uniformity and no cross-contamination between different cartridges,and the limit of detection(LOD)of this system for HBV in serum is 50 IU/*** detections of severe acute respiratory syndrome coronaviruses 2(SARS-CoV-2)with a concentration of 500 copies/mL were carried out on the system and 100%positive detection rate was achieved.

关键词： nucleic acid detection Cartridge Magnetic nanoparticles Fluidic control Point-of-care testing

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Clinical application of combined detection of SARS-CoV-2-specific antibody and nucleic acid

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World Journal of Clinical Cases 2020年第19期8卷 4360-4369页

作者： Qing-Bin Me ng Jing-Jing Peng Xin Wei Jia-Yao Yang Peng-Cheng Li Zi-Wei Qu Yong-Fen Xiong Guang-Jiang Wu Zhi-Min Hu Jian-Chun Yu Wen Su Department of Pulmonary and Critical Care Medicine and Department of Gastrointestinal Surgery Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of Respiratory Medicine General Hospital of the Yangtze River ShippingWuhan 430015Hubei ProvinceChina Department of Gastrointestinal Surgery Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430000Hubei ProvinceChina Department of Gastroenterology Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of Transfusion Medicine Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of Gastrointestinal Surgery Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of Infection Management and Disease Control Beijing Shijitan HospitalCapital Medical UniversityBeijing 100038China Department of Clinical Microbiology and Molecular Biology Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina Department of General Surgery Peking Union Medicine College HospitalChinese Academy of Medical SciencesBeijing 100730China Department of Science and Education Wuhan Integrated TCM and Western Medicine HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022Hubei ProvinceChina

BACKGROUND The global outbreak of human severe acute respiratory syndrome coronavirus(SARS-CoV)-2 infection represents an urgent need for readily available,accurate and rapid diagnostic *** acid testing of respiratory tract specimens for SARS-CoV-2 is the current gold standard for diagnosis of coronavirus disease 2019(COVID-19).However,the diagnostic accuracy of reverse transcription polymerase chain reaction(RT-PCR)tests for detecting SARS-CoV-2 nucleic acid may be lower than *** detection of SARS-CoV-2-specific antibodies should be used as a serological non-invasive tool for the diagnosis and management of SARS-CoV-2 *** To investigate the diagnostic value of SARS-CoV-2 IgM/IgG and nucleic acid detection in *** We retrospectively analyzed 652 suspected COVID-19 patients,and 206 non-COVID-19 patients in Wuhan Integrated TCM and Western Medicine *** on SARS-CoV-2 nucleic acid tests and serum antibody tests were collected to investigate the diagnostic value of nucleic acid RT-PCR test kits and immunoglobulin(Ig)M/IgG antibody test *** j2 test was used to compare differences between categorical variables.A 95%confidence interval(CI)was provided by the Wilson score *** analyses were performed with IBM SPSS Statistics version 22.0(IBM Corp.,Armonk,NY,United States).RESULTS Of the 652 suspected COVID-19 patients,237(36.3%)had positive nucleic acid tests,311(47.7%)were positive for IgM,and 592(90.8%)were positive for *** was a significant difference in the positive detection rate between the IgM and IgG test groups(Pnucleic acid test results,366 had positive IgM/IgG tests with a positive detection rate of 88.2%.CONCLUSION Our data indicate that serological IgM/IgG antibody co

关键词： SARS-CoV-2 COVID-19 nucleic acid detection Immunoglobulin M Immunoglobulin G Diagnosis

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Recent progress on rapid diagnosis of COVID-19 by point-of-care testing platforms

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Chinese Chemical Letters 2024年第3期35卷 66-75页

作者： Ya-Nan Tang Dingding Jiang Xuejun Wang Yunqi Liu Dacheng Wei State Key Laboratory of Molecular Engineering of Polymers Department of Macromolecular ScienceFudan UniversityShanghai 200433China Institute of Molecular Materials and Devices Fudan UniversityShanghai 200433China

The outbreak of COVID-19 has drawn great attention around the ***-CoV-2 is a highly infectious virus with occult transmission by many mutations and a long incubation *** particular,the emergence of asymptomatic infections has made the epidemic even more ***,early diagnosis and timely management of suspected cases are essential measures to control the spread of the *** simple,portable,and accurate diagnostic techniques for SARS-CoV-2 is the key to epidemic *** advantages of point-of-care testing technology make it play an increasingly important role in viral detection and *** review summarizes the point-of-care testing platforms developed by nucleic acid detection,immunological detection,and nanomaterial-based biosensors ***,this paper provides a prospect for designing future highly accurate,cheap,and convenient SARS-CoV-2 diagnostic technology.

关键词： COVID-19 Point-of-care testing platforms nucleic acid detection Asymptomatic infection Biosensors

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A field-deployable method for single and multiplex detection of DNA or RNA from pathogens using Cas12 and Cas13

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Science China(Life Sciences) 2022年第7期65卷 1456-1465页

作者： Lina Li Canxing Duan Jianfeng Weng Xiantao Qi Changlin Liu Xinhai Li Jinjie Zhu Chuanxiao Xie National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop ScienceChinese Academy of Agricultural Sciences(CAAS)Beijing 100081 China Biotechnology Research Institute Chinese Academy of Agricultural Sciences(CAAS)Beijing 100081China

For some Cas nucleases,trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic *** single and multiplex nucleic acid-based detection is needed for crop pathogen management in ***,we harnessed and characterized RfxCas13d as an additional CRISPR/Cas nucleic acid detection *** systematically characterized AsCas12a,LbCas12a,LwaCas13a,and RfxCas13d combined with isothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen *** data indicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as *** this tool,we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streaked dwarf virus in crude extracts prepared in the *** method,from sample preparation to result readout,could be rapidly and easily deployed in the *** system could be extended to other crop pathogens,including those that currently lack a detection method and have metabolite profiles that make detection *** nucleic acid detection system could also be used for single-nucleotide polymorphism genotyping,transgene detection,and qualitative detection of gene expression in the field.

关键词： nucleic acid detection AsCas12a LbCas12a LwaCas13a RfxCas13d maize ear rot Fusarium head blight rice black-streaked dwarf virus(RBSDV)

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