JOURNAL OF HAZARDOUS MATERIALS
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2023
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IF 13.6
Jihua Wei,Zichun Song,Jiuying Cui,Yuanxun Gong,Qianli Tang,Kai Zhang,Xinlei Song,Xianjiu Liao
In this study, we introduce an electrochemiluminescence (ECL) sensing platform based on the "Entropy-driven triggered T7 amplification-CRISPR/Cas13a system" (EDT-Cas). This platform combines a programmable entropy-driven cycling strategy, T7 RNA polymerase, and the CRISPR/Cas13a system to amplify the determination of the SARS-CoV-2 RdRp gene. The Ti 3 C 2 T x -compliant ECL signaling molecule offers unique benefits when used with the ECL sensing platform to increase the assay sensitivity and the electrode surface modifiability. To obtain the T7 promoter, the SARS-CoV-2 RdRp gene may first initiate an entropy-driven cyclic amplification response. Then, after recognizing the T7 promoter sequence on the newly created dsDNA, T7 RNA polymerase starts transcription, resulting in the production of many single-stranded RNAs (ssRNAs), which in turn trigger the action of CRISPR/Cas13a. Finally, Cas13a/crRNA identifies the transcribed ssRNA. When it cleaves the ssRNA, many DNA reporter probes carrying -U-U- are cleaved on the electrode surface, increasing the ECL signal and allowing for the rapid and highly sensitive detection of SARS-CoV-2. With a detection limit of 7.39 aM, our method enables us to locate the SARS-CoV-2 RdRp gene in clinical samples. The detection method also demonstrates excellent repeatability and stability. The SARS-CoV-2 RdRp gene was discovered using the "Entropy-driven triggered T7 amplification-CRISPR/Cas13a system" (EDT-Cas). The developed ECL test had excellent recoveries in pharyngeal swabs and environmental samples. It is anticipated to offer an early clinical diagnosis of SARS-CoV-2 and further control the spread of the pandemic.
研究领域:
生物技术
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分子生物学
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病毒学
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诊断技术
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纳米技术
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2023
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IF 4.8
Jiaying Wang,Bin Li,Bo Yao,Yi Zhao,Kai Zhang
In this work, we propose a Ti 3 C 2 T x -based electrochemiluminescence (ECL) sensing platform, combined with a programmable entropy-driven cycling strategy and inverted DNA tetrahedral nanomachines to detect the SARS-CoV-2 RdRp gene. The use of Ti 3 C 2 T x in the ECL sensing platform provides unique advantages that improve the assay's sensitivity. Firstly, the SARS-CoV-2 RdRp gene triggers an entropy-driven cyclic amplification reaction to obtain a large number of Output strands. Subsequently, the Output strand reacts with an inverted DNA tetrahedral swing arm strand to start the self-driven process. Finally, the ferrocene (Fc)-labeled hairpin DNA strand of the inverted DNA tetrahedron is severed to achieve the signal change. Under optimal conditions, the ECL platform integrated with entropy-driven cycle amplification reaction and inverted DNA tetrahedral nanomachines obtained a wide range of 100 aM to 10 nM and a limit of detection (LOD) of 48.3 aM for the SARS-CoV-2 RdRp gene. Moreover, the recovery rates of 97% to 103.3% and 98.34% to 103% were obtained in real environments and human pharyngeal swabs, respectively. This ECL sensing platform, which combines an entropy-driven cycle amplification reaction and inverted DNA tetrahedral nanomachines, opens up new opportunities for the detection of other substances.
研究领域:
生物化学
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纳米技术
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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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DOI
高影响因子
TALANTA
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2024
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IF 5.6
Jing-bo Jiao,Qing Kang,Shu-xin Cui,Jiang-li Cao,Tong Lin,Chen-jing Ma,Ze-hui Xiao,Ting Du,Nan Wang,Xin-jun Du,Shuo Wang
Coronavirus disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused secondary pandemic, which still poses a serious threat to physical health and economic development. Herein, the target-driven functionalized DNA hydrogel capillary sensor based on cascade signal amplification and carbon coated cobalt manganese modified by prussian blue and platinum nanoparticles (MnCo@C-Pt-PB NPs) has been successfully developed for dual-mode detection of SARS-CoV-2. The cascade signal amplification triggered by target RNA causes the permeability of the DNA hydrogel loaded in the capillary to be destroyed, thereby releasing the embedded MnCo@C-Pt-PB NPs as signal molecules into 3,3′,5,5′-tetramethylbenzidine/hydrogen peroxide (TMB/H 2 O 2 ) solution under the driving of capillarity. The colorless TMB is then catalyzed to blue oxidation products (oxTMB) due to peroxidase-like activity of MnCo@C-Pt-PB NPs, and MnCo@C-Pt-PB NPs and oxTMB with photothermal properties synergistically increase the system temperature under near-infrared irradiation, which are recorded by portable devices to achieve dual-mode detection. Signals intensity are proportional to the logarithm of T-RNA concentration in a wide detection range (100 aM-100 pM), with a detection limit of 100 aM. Moreover, the reliability of the developed method in oropharyngeal swabs samples has also been validated. The signal conversion and amplification function of functionalized DNA hydrogel enhances the convenience, sensitivity and versatility of the developed method, which is promising to be applied in environmental safety, molecular diagnostic assays and disease prevention.
研究领域:
生物技术
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分析化学
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纳米技术
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医学诊断
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公共卫生
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DOI
PubMed
高影响因子
Small
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2023
·Vol. 19
·期 20
·2208167
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IF 13.3
Mengya He,Xican Xu,Hongyu Wang,Qingyuan Wu,Linghao Zhang,Dongsheng Zhou,Yigang Tong,Xin Su,Huiyu Liu
Fluorescence-based PCR and other amplification methods have been used for SARS-CoV-2 diagnostics, however, it requires costly fluorescence detectors and probes limiting deploying large-scale screening. Here, a cut-price colorimetric method for SARS-CoV-2 RNA detection by iron manganese silicate nanozyme (IMSN) is established. IMSN catalyzes the oxidation of chromogenic substrates by its peroxidase (POD)-like activity, which is effectively inhibited by pyrophosphate ions (PPi). Due to the large number of PPi generated by amplification processes, SARS-CoV-2 RNA can be detected by a colorimetric readout visible to the naked eye, with the detection limit of 240 copies mL −1 . This conceptually new method has been successfully applied to correctly distinguish positive and negative oropharyngeal swab samples of COVID-19. Colorimetric assay provides a low-cost and instrumental-free solution for nucleic acid detection, which holds great potential for facilitating virus surveillance.
研究领域:
生物技术
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分子生物学
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病毒学
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诊断方法
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2022
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IF 16.744
Qizhi Liang,Yue Huang,Minghui Wang,Deqi Kuang,Jiahua Yang,Yongxiang Yi,Hai Shi,Jinlong Li,Jie Yang,Genxi Li
The persistent coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is still infecting hundreds of thousands of people every day. Enriching the kits for SARS-CoV-2 detection and developing the drugs for patient treatments are still urgently needed for combating the spreading virus, especially after the emergence of various mutants. Herein, an electrochemical biosensor has been fabricated in this work for the detection of SARS-CoV-2 via its papain-like cysteine protease (PLpro) and the screening of protease inhibitor against SARS-CoV-2 by using our designed chimeric peptide-DNA (pDNA) nanoprobes. Utilizing this biosensor, the sensitive and specific detection of SARS-CoV-2 PLpro can be conducted in complex real environments including blood and saliva. Five positive and five negative patient throat swab samples have also been tested to verify the practical application capability of the biosensor. Moreover, we have obtained a detection limit of 27.18 fM and a linear detection range from 1 pg mL −1 to 10 μg mL −1 (I = 1.63 + 4.44 lgC). Meanwhile, rapid inhibitor screening against SARS-CoV-2 PLpro can be also obtained. Therefore, this electrochemical biosensor has the great potential for COVID-19 combating and drug development.
研究领域:
生物技术
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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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药物开发
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）