ACS Sensors
·
2023
·
IF 8.9
Chenglong Lin,Zhenzhen Liu,Fanghao Fang,Shuai Zhao,Yanyan Li,Meimei Xu,Yusi Peng,Hongyou Chen,Fang Yuan,Wanju Zhang,Xi Zhang,Zheng Teng,Rui Xiao,Yong Yang
The coronavirus disease 2019 (COVID-19) pandemic highlighted the need for rapid and accurate viral detection at the point-of-care testing (POCT). Compared with nucleic acid detection, lateral flow immunoassay (LFIA) is a rapid and flexible method for POCT detection. However, the sensitivity of LFIA limits its use for early identification of patients with COVID-19. Here, an innovative surface-enhanced Raman scattering (SERS)-LFIA platform based on two-dimensional black phosphorus decorated with Ag nanoparticles as important antigen-capturing and Raman-signal-amplification unit was developed for detection of SARS-CoV-2 variants within 5–20 min. The novel SERS-LFIA platform realized a limit of detection of 0.5 pg/mL and 100 copies/mL for N protein and SARS-CoV-2, demonstrating 1000 times more sensitivity than the commercial LFIA strips. It could reliably detect seven different SARS-CoV-2 variants with cycle threshold (Ct) < 38, with sensitivity and specificity of 97 and 100%, respectively, exhibiting the same sensitivity with q-PCR. Furthermore, the detection results for 48 SARS-CoV-2-positive nasopharyngeal swabs (Ct = 19.8–38.95) and 96 negative nasopharyngeal swabs proved the reliability of the strips in clinical application. The method also had good specificity in double-blind experiments involving several other coronaviruses, respiratory viruses, and respiratory medications. The results showed that the innovative SERS-LFIA platform is expected to be the next-generation antigen detection technology. The inexpensive amplification-free assay combines the advantages of rapid low-cost POCT and highly sensitive nucleic acid detection, and it is suitable for rapid detection of SARS-CoV-2 variants and other pathogens. Thus, it could replace existing antigens and nucleic acids to some extent.
研究领域:
医学诊断
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病毒学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
APPLIED PHYSICS LETTERS
·
2023
·Vol. 123
·期 8
·
IF 4
Zhang Qianwen,Wu Huaxiong,Du Yuxiang,Zhang Minghui,Xu Bangqiang,Chen Genqiang,He Shi,Zhang Dan,Li Qi,Wang Hong-Xing
In this study, Au nanoparticles were introduced to functionalize hydrogen-terminated diamond (H-diamond) to optimize surface modification and improve the performance of diamond sensor for trace detection of SARS-CoV-2 N-protein. The Au nanoparticles were obtained by an electron beam evaporation system and annealing. Atom force microscope and scanning electron microscope were utilized to characterize the structure, morphology, and distribution of fabricated Au nanoparticles. The concentration of SARS-CoV-2 N-protein was determined by the investigation of transfer characteristics of a diamond solution gate field effect transistor. Sensitivity of this sensor was obtained as 27.43 mV/lg (N-protein concentration), which was higher than a regular H-diamond sensor. In addition, it had a wider linear detection range of 10 −15 –10 −5 mg/ml and a lower limit of detection of 10 −15 mg/ml. Accordingly, the detection performance of diamond sensor has been greatly improved due to the functionalization of Au nanoparticles.
研究领域:
生物传感器
·
材料科学
·
病毒学
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DOI
高影响因子
ANALYST
·
2025
·
IF 3.3
Yan Liu,Xue Kong,Yi Zhang,Xiumei Zhou,Zheng-Zhi Yin
A novel electrochemical microsensor is constructed on a traditional acupuncture needle (AN) and used to monitor a biomarker of the SARS-CoV-2-N protein. The reversible interaction of the borate bond between the cis-diol in this glycoprotein and the phenylboronic acid in 4-mercaptophenylboronic acid (4-MPBA) is effectively exerted. This interaction was applied to anchor the SARS-CoV-2-N protein onto 4-MPBA, which is covalently self-assemblied onto electrodeposited AuNPs by the S-Au bond. Meldola blue was then electropolymerized around the protein template. After the template was eluting, three-dimensional nanocavities complementary with the protein were generated within the polymelola blue (pMB) layer. Interestingly, nanocavities could play a channel role for the electron-transfer of outer [Fe(CN)6]3−/4−, and the signal of electrochemical probe could be hindered after recombining the SARS-CoV-2-N protein, which laid a platform for the detection of this biomarker. After optimizing the influencing factors, the prepared microsensor exhibits a linear range of 0.1 ~ 1000 ng mL−1 with a low detection limit of 0.01 ng mL−1 (S/N = 3). In particular, the sensing ability was dramatically affected by the thickness correlative factors for the polymer matrix. The suitable thickness is effective for the sensing signals, which corresponds to the behavior of surface-imprinted polymer. The microsensor showed comparatively high sensitivity and selectivity and practically detected the SARS-CoV-2-N protein in the serum sample, which is of scientific significance for the development of the electrochemical microsensor and acupuncture.
研究领域:
生物医学工程
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分析化学
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病毒学
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DOI
PubMed
高影响因子
Biosensors-Basel
·
2023
·Vol. 13
·期 7
·753
·
IF 5.4
Chonghui Yang,Yujing Yang,Gaozhen Zhao,Huan Wang,Yang Dai,Xiaowen Huang
The sensitive and rapid detection of microsamples is crucial for early diagnosis of diseases. The short response times and low sample volume requirements of microfluidic chips have shown great potential in early diagnosis, but there are still shortcomings such as complex preparation processes and high costs. We developed a low-cost smartphone-based fluorescence detection device (Smartphone-BFDD) without precision equipment for rapid identification and quantification of biomarkers on glass capillary. The device combines microfluidic technology with RGB image analysis, effectively reducing the sample volume to 20 μL and detection time to only 30 min. For the sensitivity of the device, we constructed a standard sandwich immunoassay (antibody–antigen–antibody) in a glass capillary using the N-protein of SARS-CoV-2 as a biological model, realizing a low limit of detection (LOD, 40 ng mL−1). This device provides potential applications for different biomarkers and offers wide use for rapid biochemical analysis in biomedical research.Keywords:smartphone;fluorescence;glass capillary;biomarkers;RGB
研究领域:
生物医学工程
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分析化学
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微流控
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即时诊断
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DOI
PubMed
高影响因子
Molecular Medicine Reports
·
2024
·Vol. 29
·期 3
·1-12
·
IF 3.4
Mi-Hua Liu,Xiao-Long Lin,Le-Le Xiao
Hyperphosphatemia or severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) infection can promote cardiovascular adverse events in patients with chronic kidney disease. Hyperphosphatemia is associated with elevated inflammation and sterol regulatory element binding protein 2 (SREBP2) activation, but the underlying mechanisms in SARS‑CoV‑2 that are related to cardiovascular disease remain unclear. The present study aimed to elucidate the role of excess inorganic phosphate (PI) in SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and the underlying mechanisms in vascular smooth muscle cells (VSMCs). The expression levels of SARS‑CoV‑2 N protein, SREBP cleavage‑activating protein (SCAP), mature N‑terminal SREBP2, NLRP3, procaspase‑1, cleaved caspase‑1, IL‑1β and IL‑18 were examined by western blotting. The expression levels of SREBP2, HMG‑CoA reductase, HMGCS1, low density lipoprotein receptor, proprotein convertase subtilisin/kexin type 9 (PCSK9), SREBP1c, fatty acid synthase, stearyl coenzyme A desaturase 1, acetyl‑CoA carboxylase α and ATP‑citrate lyase were determined by reverse transcription‑quantitative PCR. The translocation of SCAP or NLRP3 from the endoplasmic reticulum to the Golgi was detected by confocal microscopy. The results showed that excess PI promoted SCAP‑SREBP and NLRP3 complex translocation to the Golgi, potentially leading to NLRP3 inflammasome activation and lipogenic gene expression. Furthermore, PI amplified SARS‑CoV‑2 N protein‑induced inflammation via the SCAP‑SREBP pathway, which facilitates NLRP3 inflammasome assembly and activation. Inhibition of phosphate uptake with phosphonoformate sodium alleviated NLRP3 inflammasome activation and reduced SREBP‑mediated lipogenic gene expression in VSMCs stimulated with PI and with SARS‑CoV‑2 N protein overexpression. Inhibition of SREBP2 or small interfering RNA‑induced silencing of SREBP2 effectively suppressed the effect of PI and SARS‑CoV‑2 N protein on NLRP3 inflammasome activation and lipogenic gene expression. In conclusion, the present study identified that PI amplified SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and lipogenic gene expression via the SCAP‑SREBP signaling pathway.
研究领域:
心血管疾病
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传染病
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Junru Sun,Ruiqin Zhu,Mengxiang Wang,Jinxing Song,Lei Zhou,Zhuoya Sun,Yanze Li,Liuyang Jiao,Lu Xia,Hua He,Gaiping Zhang,Yanan Wu
Porcine epidemic diarrhea (PED) causes significant economic losses to the pig farming industry worldwide and currently lacks an effective vaccine. Multiple detection assays and protein purification methods use magnetic nanoparticles due to their biocompatibility, high specific surface area, and solution suspension properties. In this study, a two-probe competitive ELISA based on magnetic nanoparticles for detecting PEDV N protein was developed. MNPs-N and McAb-HRP probes were prepared and the procedure was optimized to identify the ideal reaction conditions. Compared to other methods, the developed method shortens the detection time to 50 min. The coefficient of variation (CV) for intra- and inter-lot replicates was less than 10 %, with reproducibility. The coincidence rate with commercial kits is 93.07 %, making this method reliable and suitable for PED immune monitoring and diagnostics.
研究领域:
兽医学
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免疫学
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分子生物学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2024
·Vol. 15
·期 1
·1-23
·
IF 14.7
Luo Yu-Wei,Zhou Jiang-Peng,Ji Hongyu,Xu Doudou,Zheng Anqi,Wang Xin,Dai Zhizheng,Luo Zhicheng,Cao Fang,Wang Xing-Yue,Bai Yunfang,Chen Di,Chen Yueming,Wang Qi,Yang Yaying,Zhang Xinghai,Chiu Sandra,Peng Xiaozhong,Huang Ai-Long,Tang Kai-Fu
Though RNAi and RNA-splicing machineries are involved in regulating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication, their precise roles in coronavirus disease 2019 (COVID-19) pathogenesis remain unclear. Herein, we show that decreased RNAi component (Dicer and XPO5) and splicing factor (SRSF3 and hnRNPA3) expression correlate with increased COVID-19 severity. SARS-CoV-2 N protein induces the autophagic degradation of Dicer, XPO5, SRSF3, and hnRNPA3, inhibiting miRNA biogenesis and RNA splicing and triggering DNA damage, proteotoxic stress, and pneumonia. Dicer, XPO5, SRSF3, and hnRNPA3 knockdown increases, while their overexpression decreases, N protein-induced pneumonia’s severity. Older mice show lower expression of Dicer, XPO5, SRSF3, and hnRNPA3 in their lung tissues and exhibit more severe N protein-induced pneumonia than younger mice. PJ34, a poly(ADP-ribose) polymerase inhibitor, or anastrozole, an aromatase inhibitor, ameliorates N protein- or SARS-CoV-2-induced pneumonia by restoring Dicer, XPO5, SRSF3, and hnRNPA3 expression. These findings will aid in developing improved treatments for SARS-CoV-2-associated pneumonia.
研究领域:
分子生物学
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病毒学
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免疫学
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药理学
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病理学
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产品关联
DOI
PubMed
高影响因子
Sensors and Actuators Reports
·
2025
·
IF 7.6
Dinglin Fan,Zhen Ren,Yongli Li,Xukui Li,Zhipiao Tian,Wei Huang,Lin Zhan,Shijia Ding,Yongjie Xu,Fuxun Yu
Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract infections in infants and children, and early and accurate diagnosis is crucial for its effective treatment and disease management. Here, we developed a nanozyme-enhanced lateral flow immunoassay (LFIA) that employed a newly generated monoclonal antibody against the conserved RSV nucleocapsid (N) protein. The platform was based on a ternary metal–organic framework nanozyme composed of iron, cobalt, and nickel ions, which exhibited synergistically enhanced peroxidase-like activity. This nanozyme efficiently catalyzed the oxidation of 3,3′ -diaminobenzidine, producing a colorimetric signal on the LFIA strip for visual detection. Due to the catalytic enhancement, the assay achieved a detection limit of 5 pg/mL for recombinant RSV N protein. For live RSV, the biosensor demonstrated a detection threshold of 10 0.875 TCID50/mL, representing a 1000-fold sensitivity improvement over AuNP-based LFIA. Moreover, the strip demonstrated high specificity and no cross-reactivity with other viruses. Owing to its simplicity, speed, and efficiency, the developed LFIA offers a robust point-of-care solution for RSV diagnosis. This platform holds great potential for clinical screening and early detection of viral infections, underscoring its value in future pandemic preparedness and respiratory disease surveillance.
研究领域:
病毒学
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免疫学
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纳米医学
·
分子诊断
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转化医学
·
临床检验
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产品关联
DOI
高影响因子
Photonics
·
2024
·Vol. 11
·期 8
·708
·
IF 2.1
Mingzhong Zhang,Shanshan Xu,Peng-Cheng Guan,Yue-Jiao Zhang,Jian-Feng Li
In surface-enhanced Raman scattering (SERS) detection research, the shape, structure, surface modification, and material selection of nanoparticles can significantly impact the SERS intensity. Petal-like gap-enhanced Raman tags (P-GERTs) possess numerous sharp tips and edges, which generate localized electric field enhancements, further amplifying the electric field enhancement effect on neighboring molecules and enhancing the SERS signal. Additionally, the surface of P-GERTs can be modified with functional molecules, enabling their application in the detection of disease biomarkers. Using COVID-19 as an example, the performance of P-GERTs in disease biomarker detection was validated, demonstrating that the signal intensity of this probe can reach 55 times that of regular gold nanoparticles and 36.7 times that of smooth shell gap-enhanced Raman tags (S-GERTs). Furthermore, in combination with magnetically retrievable magnetic bead substrates, the N-protein antigen was specifically detected in a one-step process. N-protein was detected within 15 min using a portable Raman spectrometer. The limit of detection (LOD) for the standard sample was 4.28 pg/mL, and the LOD for the actual throat swab sample system was 25.4 pg/mL. This workflow can be extended to the detection of other biomarkers, making it widely applicable.
研究领域:
分析化学
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生物医学工程
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纳米技术
·
分子生物学
·
诊断医学
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产品关联
DOI
高影响因子
ACS Sensors
·
2023
·
IF 8.9
Shixian Liu,Yuting Hou,Zirui Li,Chenyu Yang,Guozhen Liu
A fully integrated device for salivary detection with a sample-in-answer-out fashion is critical for noninvasive point-of-care testing (POCT), especially for the screening of contagious disease infection. Microfluidic paper-based analytical devices (μPADs) have demonstrated their huge potential in POCT due to their low cost and easy adaptation with other components. This study developed a generic POCT platform by integrating a centrifugal microfluidic disc with μPADs to realize sample-to-answer salivary diagnostics. Specifically, a custom centrifugal microfluidic disc integrated with μPADs is fabricated, which demonstrated a high efficiency in saliva treatment. To demonstrate the capability of the integrated device for salivary analysis, the SARS-CoV-2 Nucleocapsid (N) protein, a reliable biomarker for SARS-CoV-2 acute infection, is used as the model analyte. By the chemical treatment of the μPAD surface, and by optimizing the protein immobilization conditions, the on-disc μPADs were able to detect the SARS-CoV-2 N protein down to 10 pg mL–1 with a dynamic range of 10–1000 pg mL–1 and an assay time of 8 min. The integrated device was successfully used for the quantification of the N protein of pseudovirus in saliva with high specificity and demonstrated a comparable performance to the commercial paper lateral flow assay test strips.
研究领域:
微流控技术
·
即时检测
·
诊断学
·
生物医学工程
·
病毒学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Yi Liu,Huanjiao Weng,Zhiwei Chen,Ming Zong,Shubin Fang,Zili Wang,Shaohua He,Yangming Wu,Jizhen Lin,Shangyuan Feng,Duo Lin
The Omicron variants of SARS-CoV-2 have been spreading globally and has never disappeared from our sight, indicating that its coexistence with humans has become a fact, and monitoring its evolution and spread remains a current task. Although polymerase chain reaction (PCR) is the most commonly used virus detection method, it requires labor-intensive and time-consuming procedures in a laboratory setting. Herein, a multichannel nanoplasmonic sensing chip based on surface enhanced Raman spectroscopy (SERS) was developed for detecting N and S proteins, as well as IgG and IgM, related to SARS-CoV-2 Omicron variants. Through a self-screening process, specific antibodies for on-site and rapid identification of important variants of concerns (VoCs) were obtained, and their binding was confirmed by protein structure analysis. The use of these S protein specific antibodies can accurately identify Omicron VoCs (BA. 5, BF.7，XBB.1.5) with the detection limit (LoD) at 0.16 pg/mL. Then, the proposed SERS array chip was integrated with a hand-held Raman spectrometer to successfully detect the Omicron subvariants in real saliva samples within only 20 minutes, greatly reducing the detection time of PCR. This sensing technology will provide a powerful and rapid point-of-care testing (POCT) method for virus diagnosis, subtype identification, and post infection antibody level monitoring.
研究领域:
生物医学工程
·
分析化学
·
病毒学
·
医学诊断
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF GENERAL VIROLOGY
·
2024
·Vol. 105
·期 5
·001996
·
IF 3.6
Yuting Duan,Haichuan Li,Shuai Huang,Yaoming Li,Shuyi Chen,Lilan Xie
Transmissible gastroenteritis virus (TGEV), an enteropathogenic coronavirus, has caused huge economic losses to the pig industry, with 100% mortality in piglets aged 2 weeks and intestinal injury in pigs of other ages. However, there is still a shortage of safe and effective anti-TGEV drugs in clinics. In this study, phloretin, a naturally occurring dihydrochalcone glycoside, was identified as a potent antagonist of TGEV. Specifically, we found phloretin effectively inhibited TGEV proliferation in PK-15 cells, dose-dependently reducing the expression of TGEV N protein, mRNA, and virus titer. The anti-TGEV activity of phloretin was furthermore refined to target the internalization and replication stages. Moreover, we also found that phloretin could decrease the expression levels of proinflammatory cytokines induced by TGEV infection. In addition, we expanded the potential key targets associated with the anti-TGEV effect of phloretin to AR, CDK2, INS, ESR1, ESR2, EGFR, PGR, PPARG, PRKACA, and MAPK14 with the help of network pharmacology and molecular docking techniques. Furthermore, resistant viruses have been selected by culturing TGEV with increasing concentrations of phloretin. Resistance mutations were reproducibly mapped to the residue (S242) of main protease (Mpro). Molecular docking analysis showed that the mutation (S242F) significantly disrupted phloretin binding to Mpro, suggesting Mpromight be a potent target of phloretin. In summary, our findings indicate that phloretin is a promising drug candidate for combating TGEV, which may be helpful for developing pharmacotherapies for TGEV and other coronavirus infections.
研究领域:
病毒学
·
药理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）