ANALYTICA CHIMICA ACTA
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2024
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IF 5.7
Tianyi Zhang,Xiaoqin Wang,Yingtao Jiang,Yunyun Zhang,Shuhao Zhao,Jian Hu,Fei Hu,Niancai Peng
Background Nucleic acid testing is the most effective detection method currently available for the diagnosis of respiratory infectious diseases. However, the conventional real-time fluorescent quantitative PCR technique, which is regarded as the gold standard method for nucleic acid detection, presents significant challenges for implementation in home self-testing and popularization in underdeveloped regions due to its rigorous experimental standards. It is therefore clear that an easy-to-use, miniaturized nucleic acid testing technology and products for nonprofessionals are of great necessity to define the pathogens and assist in controlling disease transmission. (87) Results In this study, we propose a strategy for self-testing of respiratory pathogen nucleic acid that is oriented towards the public and user-friendly. The proposed system integrates the processes of extraction-free nucleic acid release, RPA isothermal amplification, and CRISPR fluorescence detection into a compact configuration. A microfluidic testing chip actuated by air pouches and a battery/USB-powered reusable device has been developed to enable simultaneous detection of internal reference genes and viral targets in a fully enclosed condition. The system allows for sample-in, and result-out testing in less than 30 minutes with a detection limit of 2 copies/μL. Additionally, a straightforward signal-light-based result display method has been developed to make it easy and intuitive for users to access the results. Furthermore, freeze-drying reagent is introduced to guarantee the storage and transportation of testing chips in ambient conditions. (135) Significance This work presents a miniaturized, portable, and highly sensitive nucleic acid detection system, where simple operating procedures have been designed for unskilled users. It is our belief that the testing system developed in this work is well suited for home-based self-testing and infection diagnosis in resource-limited areas, due to the above-mentioned advantages. (52)
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Biosensors-Basel
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2024
·Vol. 14
·期 3
·135
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IF 4.9
Hui Chen,Zhiyuan Zhuang,Naihan Xu,Ying Feng,Kaixin Fang,Chunyan Tan,Ying Tan
The ongoing COVID-19 pandemic, driven by persistent SARS-CoV-2 transmission, threatens human health worldwide, underscoring the urgent need for an efficient, low-cost, rapid SARS-CoV-2 detection method. Herein, we developed a point-of-care SARS-CoV-2 detection method incorporating recombinase polymerase amplification (RPA) and DNA–protein crosslinking chemiluminescence (DPCL) (RPADPCL). RPADPCL involves the crosslinking of biotinylated double-stranded RPA DNA products with horseradish peroxidase (HRP)-labeled streptavidin (SA-HRP). Modified products are captured using SA-labeled magnetic beads, and then analyzed using a chemiluminescence detector and smartphone after the addition of a chemiluminescent substrate. Under optimal conditions, the RPADPCL limit of detection (LOD) was observed to be 6 copies (within the linear detection range of 1–300 copies) for a plasmid containing the SARS-CoV-2 N gene and 15 copies (within the linear range of 10–500 copies) for in vitro transcribed (IVT) SARS-CoV-2 RNA. The proposed method is convenient, specific, visually intuitive, easy to use, and does not require external excitation. The effective RPADPCL detection of SARS-CoV-2 in complex matrix systems was verified by testing simulated clinical samples containing 10% human saliva or a virus transfer medium (VTM) spiked with a plasmid containing a SARS-CoV-2 N gene sequence or SARS-CoV-2 IVT RNA. Consequently, this method has great potential for detecting targets in clinical samples.
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TALANTA
·
2024
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IF 5.6
Bing Zhao,Chan-Ru Xiong,Yao Liu,Qing-Cai Yu,Xing Chen
The outbreak of novel coronavirus pneumonia (COVID-19) in 2019 has garnered widespread attention. The virus exhibits high contagiousness, and in certain cases, it can lead to recurrent infections. Therefore, it is imperative to develop portable, sensitive, and accurate sensors to promptly detect infected individuals, control the virus's transmission, and determine suitable treatment strategies. In this study, we proposed a magnetically-assisted method employing CFO@CS-Au MNP as the substrate material, which was functionalized with human angiotensin-converting enzyme (ACE2) for efficient capture of SARS-CoV-2 spike protein in solution. Subsequently, the captured protein was sensitively detected through differential pulse voltammetry (DPV) electrical analysis. The linear detection range of the labeled GCE/MNP/GA/ACE2/BSA electrochemical sensor is from 1 pg/mL to 10 μg/mL, with a minimum detection limit of 0.15 pg/mL. Furthermore, the fabricated GCE/MNP/GA/ACE2/BSA sensor achieved satisfactory recoveries of SARS-CoV-2 spike protein in saliva and nasal swab samples within 10 min. These results indicate that this magnetically-assisted biosensor has established a solid foundation for the swift on-site detection of COVID-19.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Aihua Lyu,Tengchuan Jin,Shanshan Wang,Xiaoxue Huang,Weihong Zeng,Rui Yang,Hua Cui
Accurate and efficient early diagnosis is crucial for the control of COVID-19 pandemic. However, methods that can balance sensitivity, high throughput, detection speed and automation simultaneously are still scarce. Here, we report an automatic label-free chemiluminescence immunoassay (CLIA) for rapid SARS-CoV-2 nucleocapsid protein (NP) detection with high sensitivity and throughput. N-(4-aminobutyl)-N-ethylisoluminol and Co 2+ dual-functionalized chemiluminescent magnetic beads (dfCL-MB) were first applied to the detection of protein by a novel and simple strategy. Sulphydryl polyethylene glycol was coated on the surface of dfCL-MB so as to assemble dfCL-MB and antibody conjugated gold nanoparticles through Au-S bond. Considering the high-risk application scenarios, the immunosensor was integrated with an automatic chemiluminescence analyzer so that the whole testing procedure could be carried out automatically without manual operation. A linear correlation between CL intensities and the logarithm of NP concentration was obtained in the range of 0.1–10,000 pg/mL with a detection limit of 21 fg/mL. The whole process cost 25 min and the sample compartment can bear 24 samples simultaneously. The spiked human serum samples and serum samples from COVID-19 patients were determined with satisfactory recoveries of 91.1–109.4%, suggesting that the proposed label-free CLIA is of great potential for SARS-CoV-2 NP detection in practice.
研究领域:
生物医学工程
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分析化学
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分子生物学
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病毒学
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临床诊断
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Frontiers in Immunology
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2026
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IF 7
Anran Shen,Suyue Zhu,Min Li,Yu Zhao,Yandan Wu,Yue Zhang,Jiejie Zhang,Xuelian Han,Yuan Wang,Guangyu Zhao,Linli Lv,Qi Yin,Taotao Tang
Objective To evaluate the synergistic immunological protection of exosomal T-cell epitope vaccine and antibody-inducing vaccine against SARS-CoV-2 in highly humanized mice. Methods Red blood cell-derived exosomes were loaded with 27 CD8 + T-cell epitope peptides and 19 CD4 + T-cell epitope peptides followed by combined immunization with S1 protein vaccine of SARS-CoV-2 and adjuvant poly I:C in HLA-A2/DR1 double transgenic mice. After immunizations, splenocytes were assessed for epitope-specific T cell responses by intracellular cytokine staining and ELISPOT, and for functional T cell subset analysis through flow cytometry. Meanwhile, serum anti-S1 protein IgG and neutralizing antibodies were quantified via ELISA and BA.5 pseudovirus neutralization assay, respectively. Furthermore, viral challenge was performed after the combined immunization in HLA-A2/DR1/hACE2 triple transgenic mice, followed by viral load quantification, viral protein detection, and H&E staining in lungs. Results The combined immunization i) increased the titers of S1 protein-specific IgG antibodies and neutralizing antibodies as well as Tfh cell frequency as compared to the S1 protein vaccine alone; ii) induced significantly more S1 protein-specific T cells and effector memory CD4 + T cells, and inhibited T cell exhaustion and regulatory T cell differentiation, compared to the exosomal T-cell epitope vaccine alone; iii) achieved the lowest pulmonary viral loads, inflammatory cell infiltration, and histopathological damage after SARS-CoV-2 infection. Conclusion This study, for the first time, demonstrates the synergistic humoral and cellular immune responses and protective efficacy induced by the combined immunization of exosomal T-cell epitope vaccine and antibody-inducing vaccine, and provides preclinical evidence from highly humanized mice for optimizing next-generation SARS-CoV-2 vaccine protocols.
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ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
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2026
·
IF 3.5
Margarita A. Arbuzova,Olga Yu Selyutina,Nikolay E. Polyakov
Cholesterol drives E-protein depth redistribution via a matrix-mediated helix tilt adjustment. • Escin-induced peptide localization shifts strictly require the presence of charged DMPS lipids. • Escin is hypothesized to modulate the viroporin conformation via localized helix kinking rather than matrix disruption. • Escin is able to extract cholesterol from lipid membrane. β-Escin is a well-known saponin derived from the horse chestnut tree. Saponins, including escin, exhibit a wide range of biological activities, among which membrane-modifying activity is particularly notable. The latter is key to understanding the molecular mechanisms underlying escin's biological effects, including its antiviral properties. For example, altering membrane properties can strongly influence the life cycle of enveloped viruses such as SARS-CoV-2 by modulating the activity of its envelope protein (E protein). In the present paper, we studied the localization of both escin and the E protein transmembrane domain (ETM) in model membranes with different lipid compositions. We demonstrated that escin can alter ETM localization in membranes containing the negatively charged lipid DMPS, and that escin is also capable of extracting cholesterol from model lipid membranes. Download: Download high-res image (423KB) Download: Download full-size image
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Frontiers in Cellular and Infection Microbiology
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2021
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IF 5.293
Zhang Qi,Chen Zhiqiang,Huang Chenxiao,Sun Jiuyuan,Xue Minfei,Feng Tingting,Pan Wen,Wang Kezhen,Dai Jianfeng
Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread worldwide and has infected more than 250 million people. A typical feature of COVID-19 is the lack of type I interferon (IFN-I)-mediated antiviral immunity in patients. However, the detailed molecular mechanisms by which SARS-CoV-2 evades the IFN-I-mediated antiviral response remain elusive. Here, we performed a comprehensive screening and identified a set of SARS-CoV-2 proteins that antagonize the IFN-I response. Subsequently, we characterized the mechanisms of two viral proteins antagonize IFN-I production and downstream signaling. SARS-CoV-2 membrane protein binds to importin karyopherin subunit alpha-6 (KPNA6) to inhibit interferon regulatory factor 3(IRF3) nuclear translocation. Further, the spike protein interacts with signal transducer and activator of transcription 1 (STAT1) to block its association with Janus kinase 1 (JAK1). This study increases our understanding of SARS-CoV-2 pathogenesis and suggests novel therapeutic targets for the treatment of COVID-19.
研究领域:
病毒学
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免疫学
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分子生物学
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NEW JOURNAL OF CHEMISTRY
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2023
·Vol. 48
·期 4
·1502-1509
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IF 3.3
Yansheng Liu,Junpeng Deng,Xiaobo Jia,Jin Zhou,Hongli Li,Xiaohong Wang,Yating Chen,Zhenle Qin,Zhichen Jin,Guofu Wang
Quick and highly sensitive detection of the SARS-CoV-2 virus can efficiently reduce the possibility of viral spreading. In this study, a differential-phase surface plasmon resonance (SPR) biosensor is constructed to directly detect the SARS-CoV-2 virus. The SARS-CoV-2 spike protein antibodies are applied as bioprobes and functionalized on Au film, forming a sensing surface. A numerical analysis of the phase shifts of the SPR interferogram, termed the index of phase shifts (IoPS), is presented, which does not depend on a reference beam or an s-component beam. The IoPS can sensitively reflect small changes in the refractive index (as low as 3.75 × 10−10 RIU) when NaCl solution is used as an analyte. The limit of detection (LOD) of the proposed SPR biosensor in detecting the SARS-CoV-2 S protein is 1 ag mL−1. In comparison with a SARS-CoV-2 negative control (NC) sample collected from healthy people, which has an IoPS of ΔϕNC = 39°, the SPR biosensor shows a very strong response to the SARS-CoV-2 virus collected from infected people with a phase shift of ΔϕIoPS = 566°. To further improve the detection sensitivity, a sandwich structure is designed by applying SARS-CoV-2 spike protein antibody functionalized gold nanoparticles (anti-S@AuNPs). This reduces the LOD for the detection of the SARS-CoV-2 S protein to 0.1 ag mL−1.
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BIOSENSORS & BIOELECTRONICS
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2023
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IF 12.6
Hong Zhang,Chenggang Zhang,Zhaotong Wang,Wenwu Cao,Miao Yu,Ye Sun
Sensitive and anti-interference detection of targeted signal(s) in body fluids is one of the paramount tasks in biosensing. Overcoming the complication and high cost of antibody/aptamer-modification, surface-enhanced Raman spectroscopy (SERS) based on antibody/aptamer-free (AAF) substrates has shown great promise, yet with rather limited detection sensitivity. Herein, we report ultrasensitive and anti-interference detection of SARS-CoV-2 spike protein in untreated saliva by an AAF SERS substrate, applying the evanescent field induced by the high-order waveguide modes of well-defined nanorods for SERS for the first time. A detection limit of 3.6 × 10 −17 M and 1.6 × 10 −16 M are obtained in phosphate buffered saline and untreated saliva, respectively; the detection limits are three orders of magnitude improved than the best records from AAF substrates. This work unlocks an exciting path to design AAF SERS substrates for ultrasensitive biosensing, not limited to detection of viral antigens.
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MICROCHEMICAL JOURNAL
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2022
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IF 5.304
Huiqin Chang,Meng Jiang,Qiying Zhu,Anqi Liu,Yuyin Wu,Canguo Li,Xiangyue Ji,Li Gong,Shanshan Li,Zhiwei Chen,Ling Kong,Lei Han
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-COV-2) is a cluster of β coronaviruses. The 2019 coronavirus disease (COVID-19) caused by SARS-COV-2 is emerging as a global pandemic. Thus, early diagnosis of SARS-COV-2 is essential to prevent severe outbreaks of the disease. In this experiment, a novel label-free photoelectrochemical (PEC) immunosensor was obtained based on silver sulfide (Ag 2 S) sensitized titanium dioxide@bismuth tungstate (TiO 2 @Bi 2 WO 6 ) nanocomposite for quantitative detection of SARS-COV-2 nucleocapsid protein. The constructed TiO 2 @Bi 2 WO 6 hollow microspheres had large specific surface area and could produce high photocurrent intensity under visible light illumination. Ag 2 S was in-situ grown on the surface of thioglycolic acid (TGA) modified TiO 2 @Bi 2 WO 6 . In particular, TiO 2 @Bi 2 WO 6 and Ag 2 S formed a good energy level match, which could effectively enhance the photocurrent conversion efficiency and strength the photocurrent response. Ascorbic acid (AA) acted as an effective electron donor to effectively eliminate photogenerated holes. Under optimal experimental conditions, the constructed immunosensor presented a supersensitive response to SARS-COV-2 nucleocapsid protein, with a desirable linear relationship ranged from 0.001 to 50 ng/mL for nucleocapsid protein and a lower detection limit of 0.38 pg/mL. The fabricated sensor exhibited a wide linear range, excellent selectivity, specificity and stability, which provided a valuable referential idea for the detection of SARS-COV-2.
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Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Ju Jian,Zhang Xinyu,Li Lin,Regmi Sagar,Yang Guoqiang,Tang Shixing
Understanding the dynamic changes in antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential for evaluating the effectiveness of the vaccine and the stage for the recovery of the COVID-19 disease. A rapid and accurate method for the detection of SARS-CoV-2-specific antibodies is still urgently needed. Here, we developed a novel fluorescent lateral flow immunoassay (LFA) platform for the detection of SARS-CoV-2-specific IgM and IgG by the aggregation-induced emission carbon dots conjugated with the SARS-CoV-2 spike protein (SSP). The aggregation-induced emission carbon dots (AIE-CDs) are one of the best prospect fluorescent probe materials for exhibiting high emission efficiency in both aggregate and solid states. The AIE-CDs were synthesized and displayed dual fluorescence emission, which provides a new perspective for the design of a high sensitivity testing system. In this work, the novel LFA platform adopted the AIE carbon dots, which are used to detect SARS-CoV-2-specific IgM and IgG conveniently. Furthermore, this sensor had a low LOD of 100 pg/ml. Therefore, this newly developed strategy has potential applications in the areas of public health for the advancement of clinical research.
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ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Zihao Xie,Shasha Feng,Fubin Pei,Mingzhu Xia,Qingli Hao,Bing Liu,Zhaoyang Tong,Jiang Wang,Wu Lei,Xihui Mu
The SARS-CoV-2 pandemic has posed a huge challenge to rapid and accurate diagnosis of SARS-CoV-2 in the early stage of infection. In this work, we developed a novel magnetic/fluorescent dual-modal lateral flow immunoassay (LFIA) based on multifunctional nanobeads for rapid and accurate determination of SARS-CoV-2 nucleocapsid protein (NP). The multifunctional nanobeads were fabricated by using polyethyleneimine (PEI) as a mediate shell to combine superparamagnetic Fe 3 O 4 core with dual quantum dot shells (MagDQD). The core-shell structure of MagDQD label with high loading density of quantum dots (QDs) and superior magnetic content realized LFIA with dual quantitative analysis modal from the assemblies of individual single nanoparticles. The LFIA integrated the advantages of magnetic signal and fluorescent signal, resulting excellent accuracy for quantitative analysis and high elasticity of the overall detection. In addition, magnetic signal and fluorescent signal both had high sensitivity with the limit of detection (LOD) as 0.235 ng mL −1 and 0.012 ng mL −1 , respectively. The recovery rates of the methods in simulated saliva samples were 91.36%–103.60% (magnetic signal) and 94.39%–104.38% (fluorescent signal). The results indicate the method has a considerable potential to be an effective tool for diagnose SARS-CoV-2 in the early stage of infection.
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