A novel cartridge for nucleic acid extraction,amplification and detection of infectious disease pathogens with the help of magnetic nanoparticles |
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| Institution: | 1. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China;2. Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province, Hunan Key Laboratory of Green Chemistry and Application of Biological Nanotechnology, Hunan University of Technology, Zhuzhou 412007, China;1. Key Laboratory of Agricultural Information Acquisition Technology, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing 100083, China;2. Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville 72701, United States;3. College of Veterinary medicine, South China Agricultural University, Guangzhou 510642, China;1. Key Laboratory of Luminescence Analysis and Molecular Sensing, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China;2. College of Food Science, Southwest University, Chongqing 400715, China;1. Medical College, Guizhou University, Guiyang 550000, China;2. Department of Orthopedics, Guizhou Provincial People''s Hospital, Guiyang 550000, China;3. Department of Orthopaedics, The Affiliated Hospital of Guizhou Medical University, Guiyang 550000, China;1. Institute of Chemical Industry of Forest Products, Chinese Academy of Forestry; Key Lab. of Biomass Energy and Material, Jiangsu Province; National Engineering Lab for Biomass Chemical Utilization, Nanjing 210042, China;2. Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources; International Innovation Center for Forest Chemicals and Materials College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China;3. Institute of Nanochemistry and Nanobiology School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444, China;1. CAS Key Laboratory of Environmental and Applied Microbiology, Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China;2. College of Resources and Environmental Engineering, Mianyang Normal University, Mianyang 621000, China;3. Department of Materials and Chemical Engineering, Yibin University, Yibin 644000, China;1. School of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou 225002, China;2. College of Environmental Science and Engineering, Yangzhou University, Yangzhou 225002, China |
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| Abstract: | Nucleic acid detection (NAD) based on real-time polymerase chain reaction (real-time PCR) is gold standard for infectious disease detection. Magnetic nanoparticles (MNPs) are widely used for nucleic acid extraction (NAE) because of their excellent properties. Microfluidic technology makes automated NAD possible. However, most of the NAD microfluidic chips are too complex to be applied to point-of-care (POC) testing. In this paper, a simple-structure cartridge was developed for POC detection of infectious diseases. This 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 500 µL of liquid samples by MNPs and finish the detection process from “sample in” to “answer out” automatically. Performance tests of the cartridges show that it met the demands of automated NAD. Results 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/mL. Multiplex 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. |
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