TALANTA
·
2023
·
IF 6.1
Haodong Cui,Wenxing Song,Xiaoling Ru,Wen Fu,Ling Ji,Wenhua Zhou,Zhen Zhao,Guangbo Qu,Xue-Feng Yu,Guibin Jiang
The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based methods are being widely applied for clinical diagnosis, timely and accurate diagnosis towards COVID-19 causing virus, the SARS-CoV-2, is still limited due to labor-intensive and time-consuming operations. Herein, we report a new viral RNA extraction method based on poly-(amino ester) with carboxyl group (PC)-coated magnetic nanoparticles (pcMNPs) for the sensitive detection of SARS-CoV-2. This method combines the lysis and binding steps into one step, and refines multiple washing steps into one step, giving a turnaround time of less than 9 min. Furthermore, the extracted pcMNP-RNA complexes can be directly introduced into subsequent RT-PCR reactions without elution. This simplified viral RNA method could be well adapted in fast manual and automated high-throughput nucleic acids extraction protocols suitable for different scenarios. A high sensitivity down to 100 copies/mL and a linear correlation between 100 and 10 6 copies/mL of SARS-CoV-2 pseudovirus particles are achieved in both protocols. Benefitting from the simplicity and excellent performances, this new method can dramatically improve the efficiency and reduce operational requirements for the early clinical diagnosis and large-scale SARS-CoV-2 nucleic acid screening.
研究领域:
分子生物学
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医学诊断
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纳米技术
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传染病研究
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Xueliang Lin,Youliang Weng,Yi Liu,Duo Lin,Huishan Yang,Zhiwei Chen,Shangyuan Feng
The rapid and high precision analysis of SARS-CoV-2 RNA in saliva is crucial for the efficient prevention and control of COVID-19. Herein, we developed a novel ratiometric surface-enhanced Raman spectroscopy (SERS) sensing chip based on a two-dimensional core-molecule-shell-molecule nanostructure for quantitative detection of SARS-CoV-2 RNA. Owing to the use of base-complementary pairing mechanism, this SERS sensing chip has better selectivity in discriminating SARS-CoV-2 RNA against other RNA. Besides, an internal standard molecule (4MBN) in the SERS sensing chip can used to calibrate the real-time changing Raman signals of the probe molecule during quantitative assay. Using this cleverly designed SERS sensing chip, the dynamic ranges from 10 –6 M to 10 –12 M with a low of detection limit (LOD = 7.61 ×10 –14 M) can be acquired for detection of SARS-CoV-2 RNA. Meanwhile, this SERS sensing chip was well adapted for point-of-care testing (POCT) in combination with portable Raman spectrometer, which is expected to achieve effective detection of early SARS-CoV-2 infection.
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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高影响因子
JOURNAL OF LIGHTWAVE TECHNOLOGY
·
2024
·Vol. 42
·期 16
·5768-5774
·
IF 4.1
Xuegang Li,Huiying Sun,Yanan Zhang,Yong Zhao,Xue Zhou
In this paper, a groundbreaking probe-type fiber optic biosensor, harnessing the surface plasmon resonance (SPR) effect enhanced by barium titanate glass microspheres, is unveiled for virus RNA detection. The multi-physics simulation was used to verify the sensitivity improvement achieved by introducing barium titanate microspheres on the surface of the optical fiber microspheres probe. Experiments confirmed this, pushing the sensor's limit to an impressive 6802 nm/RIU. Biomarker capture on the microsphere surface subtly alters the local refractive index, triggering a detectable SPR spectral shift. To showcase its prowess, the sensor tackled the reverse-transcribed complementary DNA (cDNA) of COVID-19 RNA. Chemical modification of the gold film and DNA hybridization technology ensured specific cDNA detection. The limit of detection for this target reached a remarkable 1.58 nmol/L. This sensor not only excels in virus RNA detection but also holds immense potential for diagnosing major diseases by pinpointing the concentrations of key biomarkers.
研究领域:
生物医学工程
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光学传感
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分子诊断
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DOI
高影响因子
Frontiers in Microbiology
·
2020
·
IF 4.236
Hu Ao,Li Jing,Tang Wei,Liu Ge,Zhang Haiwei,Liu Chunlan,Chen Xulin
Influenza virus RNA-dependent RNA polymerase (vRdRp) does not have capping activity and relies on the capped RNAs produced by the host RNA polymerase II (RNAPII). The viral polymerases process the capped RNAs to produce short capped RNA fragments that are used as primers to initiate the transcription of viral mRNAs. This process, known as cap-snatching, can be targeted by antiviral therapeutics. Here, anthralin was identified as an inhibitor against influenza a virus (IAV) infection by targeting the cap-snatching activity of the viral polymerase. Anthralin, an FDA-approved drug used in the treatment of psoriasis, shows antiviral activity against IAV infection in vitro and in vivo. Importantly, anthralin significantly reduces weight loss, lung injury, and mortality caused by IAV infection in mice. The mechanism of action study revealed that anthralin inhibits the cap-binding function of PB2 subunit and endonuclease activity of PA. As a result, viral mRNA transcription is blocked, leading to the decreases in viral RNA replication and viral protein expression. In conclusion, anthralin has been demonstrated to have the potential of an alternative antiviral against influenza virus infection. Also, targeting the captive pocket structure that includes the N-terminus of PA endonuclease domain and the C-terminal of PB2 cap-binding domain of IAV RdRp may be an excellent strategy for developing anti-influenza drugs.
研究领域:
病毒学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Yifei Li,Jingwen Li,Ya Cao,Ping Jiang,Yuguo Tang,Zutao Chen,Kun Han
Sensitive and specific assay of hepatitis C virus (HCV) RNA has a great clinical significance for the diagnosis of hepatitis C in window period. A colorimetric assay is described for determination of the RNA of hepatitis C virus. The method is based on the in situ synthesis of graphene quantum dot-silver nano-composites (GQD/Ag NCs), magnetic separation and catalyzed hairpin assembly reaction (CHA). The silver component of GQD/Ag NCs react with H 2 O 2 generated by glucose oxidase , which are induced via CHA. Then the light-yellow solution turns into colorless transparent in the presence of the target RNAs. The method has a linear response in the 25–500 pM RNA concentration range, and the detection limit is 24.84 pM. The method is simple, stable and sensitive and might be extend to the assay of other RNA viruses . The retro-transcription free and visual distinguishable properties endow this strategy a promising application in the screening of HCV.
研究领域:
医学诊断
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分子生物学
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DOI
PubMed
高影响因子
SENSORS
·
2018
·Vol. 18
·期 9
·2980
·
IF 2.475
Zhizhong Han,Qinghua Weng,Chaofan Lin,Jinquan Yi,Jie Kang
In this work, the CdSe–ZnO flower-rod core-shell structure (CSZFRs) was prepared by ion-exchange method. The surface of CSZFRs was modified by 3-mercaptopropionic acid (MPA), and then the DNA probe was immobilized on the surface via chemical bond between -NH2of DNA probe and -COOH of MPA. Finally, the target norovirous (NV) RNA was combined with the probe according to the principle of complementary base pairing, resulting in a decrease of the photocurrent. The results show that the absorbance spectrum of visible light is enhanced for CSZFRs compared with pure ZnO. Under visible light irradiation, the photocurrent of CSZFRs is up to 0.1 mA, which can improve the sensitivity of the photoelectrochemical (PEC) biosensor. In the measurement range of 0–5.10 nM, the measured concentrations (c) have a good linear relationship with the output photocurrent of the biosensor. The linear regression equation is expressed asI= 0.03256 − 0.0033c(R2= 0.99, S/N = 3) with a detection limit of 0.50 nM. Therefore, this work realizes a rapid and sensitive method for the detection of NV RNA.Keywords:CdSe–ZnO flower-rod;photoelectrochemical biosensor;norovirous RNA;ion-exchange method
研究领域:
材料科学
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生物化学
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分析化学
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病毒学
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Jiwei Wang,Chuankun Yang,Ran Xu,Shuo Ma,Ai Chen,Yuming Yao,Fengfeng Zhao,Guoqiu Wu
This method pioneers a synergistic strategy combining ‘extended-strand unmasking with freeze-drying pre-packaging’. By designing unequal-length double-strand replacement strands that competitively bind to regions flanking the target site, it forces the unfolding of hairpin/cloverleaf conformations, thereby physically exposing concealed targets. Concurrently, a five-step lyophilization process (pre-folding-anchoring-sealing-encapsulation-freezing) employs graphene oxide (GO) π-π stacking to suppress >90 % of DNA breathing effects. Combining 0.02 % (w/v) low-melting-point agarose achieves in situ encapsulation and probe conformation fixation, forming microchannels for water molecules. The final lyophilized product forms a porous solid matrix (integrable into nitrocellulose test strips or 2-mm diameter lyophilized spheres), enabling stable probe storage at ambient temperature. This technology overcomes the ‘impossible triangle’ challenge in molecular diagnostics: achieving high room-temperature stability (>6 months, compared to <7 days for traditional liquid-phase CHA), ultra-high sensitivity (0.01 pM), and a simple, rapid testing process. This solution provides a “sample-in, result-out” point-of-care testing approach for home health management (no cold chain required), primary care clinics (no equipment dependency), and pandemic emergency response.
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ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Benyu He,Lingfeng Wang,Xinyu Jin,Xing Zhang,Rui Sha,Yong Liang,Yawei Wang,Wenjing Xie,Jianbo Shi,Hanyong Peng
The detection of virus RNA in wastewater has been established as a valuable method for monitoring Coronavirus disease 2019. Carbon nanomaterials hold potential application in separating virus RNA owing to their effective adsorption and extraction capabilities. However, carbon nanomaterials have limited separability under homogeneous aqueous conditions. Due to the stabilities in their nanostructure, it is a challenge to efficiently immobilize them onto magnetic beads for separation. Here, we develop a porous agarose layered magnetic graphene oxide (GO) nanocomposite that is prepared by agglutinating ferroferric oxide (Fe3O4) beads and GO with agarose into a cohesive whole. With an average porous size of approximately 500 nm, the porous structure enables the unhindered entry of virus RNA, facilitating its interaction with the surface of GO. Upon the application of a magnetic field, the nucleic acid can be separated from the solution within a few minutes, achieving adsorption efficiency and recovery rate exceeding 90% under optimized conditions. The adsorbed nucleic acid can then be preserved against complex sample matrix for 3 days, and quantitatively released for subsequent quantitative reverse transcription polymerase chain reaction (RT-qPCR) detection. The developed method was successfully utilized to analyze wastewater samples obtained from a wastewater treatment plant, detecting as few as 10 copies of RNA molecules per sample. The developed aMGO-RT-qPCR provides an efficient approach for monitoring viruses and will contribute to wastewater-based surveillance of community infections.
研究领域:
环境科学
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纳米技术
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病毒学
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分析化学
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DOI
PubMed
高影响因子
ACS Sensors
·
2022
·
IF 9.618
Jiahao Li,Lina Tang,Tingxian Li,Kun Li,Yulin Zhang,Wei Ni,Meng-Meng Xiao,Youyun Zhao,Zhi-Yong Zhang,Guo-Jun Zhang
The path toward field-effect transistor (FET) application from laboratory to clinic has delivered a compelling push in the biomedical domain, yet ultrasensitive and timely pathogen identification without PCR remains a long-lasting challenge. Herein, we create a generic check station termed “CRISPR-FET”, first incorporating the CRISPR/Cas13a system within the FET modality, for accelerated and unamplified detection of viral RNA. Unlike conventional FETs bearing target-specific receptors, this sensor holds three unique advancements: (i) an ingenious sensing mechanism is used, which converts the signal of a large-sized analyte into an on-chip cleavage response of an immobilized CRISPR reporter, enabling signal generation events to occur all within the Debye length; (ii) the multipurpose inspection of the CoV ORF1ab, CoV N gene, and HCV RNA unveils the potential for “one-for-all” scalable FET-based molecular diagnostics; and (iii) it is shown that Cas13a-crRNAs targeting different sites of the viral genome can be deployed in tandem to amplify the FET response, empowering the detection limit down to 1.56 aM, which is a world-record level of sensitivity in the FET for direct viral gene sensing. Notably, a brilliant clinical applicability was made in distinguishing HCV-infected patients from normal controls. Overall, this study sheds new insights into FET-based nucleic acid sensing technology and invokes a vision for its possible future roles in diagnosis of various viral diseases.
研究领域:
生物医学工程
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分子诊断
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纳米技术
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病毒学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Xin Gao,Yun Yin,Junyu Xie,Shaohua Gong,Xuemei Li
CRISPR/Cas13a system has been combined with different isothermal amplification strategies for RNA analysis. Nevertheless, expensive enzymes are usually indispensable in these isothermal amplification methods, which increases the detection cost. In this study, a CRISPR/Cas13a-mediated triple signal amplification strategy has been designed for selective, sensitive and inexpensive detection of viral RNA. SARS-CoV-2 S RNA activated CRISPR/Cas13a to cut hairpin DNA, triggering catalytic hairpin assembly (CHA) and subsequent hybridization chain reaction (HCR) to form a long-stranded Y-type DNA. G-quadruplexes were distributed on the branch of the long-stranded DNA and bound with N-methyl mesoporphyrin IX (NMM) to enhance the fluorescence signal. SARS-CoV-2 S RNA was specifically detected with a limit of detection (LOD) of 285 fM. Notably, no additional expensive enzymes were required in the entire detection process. The CRISPR/Cas13a-mediated triple signal amplification strategy holds great potential for specific and sensitive viral RNA detection with low cost.
研究领域:
生物技术
·
分子生物学
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病毒学
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产品关联
DOI
高影响因子
LANGMUIR
·
2024
·
IF 3.7
Yaqin Zhang,Jianai Chen,Fange Kong,Chunxia Wang,Hangyu Guo,Yingchun Li,Jiahui Lu,Jicheng Zhang,Jiasi Wang,Yulin Zhou
Viral infections, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are some of the most dangerous threats to humans. SARS-CoV-2 has caused a global pandemic, highlighting the unprecedented demand for rapid and portable diagnostic methods. To meet these requirements, we designed a label-free colorimetric platform that combines the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) 12a system for naked-eye detection (named LFP). This method utilizes reverse transcription loop-mediated isothermal amplification (RT-LAMP) and the trans-cleavage activity of the CRISPR/Cas12a system to increase the sensitivity and specificity of the reaction. This platform can detect as few as 4 copies/μL of RNA and produces no false positive results when tested against the influenza virus. To better meet the requirements of point-of-care (POC) detection, we developed a portable device that can be applied in resource-poor and densely populated regions. The LFP assay holds great potential for application in resource-limited settings, and the label-free gold nanoparticle (AuNPs) probe can reduce costs, making it suitable for large-scale screening. We expect that the LFP assay will be promising for the POC screening of COVID-19.
研究领域:
分子生物学
·
生物技术
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病毒学
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纳米技术
·
诊断方法
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOLOGICAL CHEMISTRY
·
2021
·Vol. 297
·期 1
·
IF 5.157
Bochuan Yuan,Ziwei Wu,Wei Ji,Dong Liu,Xiang Guo,Donghui Yang,Aili Fan,Hongli Jia,Ming Ma,Wenhan Lin
The fungal cyclohexadepsipeptides destruxins (DTXs), isaridins (ISDs), and isariins (ISRs) are nonribosomal peptides whose structures include a 19-membered ring composed of five amino acid residues and one α- or β-hydroxy acid residue. These cyclohexadepsipeptides contain unusual nonproteinogenic amino acid–building blocks and possess a range of antiviral, antibacterial, and other activities. The biosynthetic gene clusters for ISDs and ISRs have not been identified, and the biosynthesis of the nonproteinogenic (3S)-methyl-l-proline residue, which is found in DTXs, ISDs, and many other natural products, lacks full characterization. In an ongoing effort to identify compounds that can inhibit the Zika virus (ZIKV), we examined the extract of marine-derived fungus Beauveria felina SX-6-22 and discovered 30 DTXs, ISDs, and ISRs (1–30) including seven new compounds (1–7). The anti-ZIKV assays showed that 9–12 and 16–18 possess inhibitory activities against ZIKV RNA replication and NS5 (nonstructural protein 5) production in ZIKV-infected A549 cells. We sequenced the genome of B. felina SX-6-22 and identified three biosynthetic gene clusters detx, isd and isr, which are responsible for the biosynthesis of DTXs, ISDs, and ISRs, respectively. Comparative analyses of the three gene clusters clarified the biosynthetic relationships among these cyclohexadepsipeptides. Finally, we characterized the entire biosynthesis of nonproteinogenic building block (3S)-methyl-l-proline. The Δ1-pyrroline-5-carboxylate reductases (P5CRs), also used in the biosynthesis of l-proline, were demonstrated to catalyze the final reduction step in (3S)-methyl-l-proline formation, suggesting potential cross talk between primary and secondary metabolisms. These results provide opportunities for biosynthetic pathway engineering to generate new anti-ZIKV cyclohexadepsipeptides.
研究领域:
药理学
·
微生物学
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分子生物学
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生物化学
·
基因组学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）