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RdRp基因
ADVANCED FUNCTIONAL MATERIALS · 2022 ·Vol. 32 ·期 48 ·2209743 · IF 19.924

2D Zn-Porphyrin-Based Co(II)-MOF with 2-Methylimidazole Sitting Axially on the Paddle–Wheel Units: An Efficient Electrochemiluminescence Bioassay for SARS-CoV-2

Yi-Xuan Li,Jing Li,Dunru Zhu,Ju-Zheng Wang,Guo-Fang Shu,Junji Li,Sheng-Li Zhang,Xue-Ji Zhang,Serge Cosnier,Hai-Bo Zeng,Dan Shan

High electrocatalytic activity with tunable luminescence is crucial for the development of electrochemiluminescence (ECL) luminophores. In this study, a porphyrin-based heterobimetallic 2D metal organic framework (MOF), [(ZnTCPP)Co 2 (MeIm)] (1), is successfully self-assembled from the zinc(II) tetrakis(4-carboxyphenyl)porphine (ZnTCPP) linker and cobalt(II) ions in the presence of 2-methylimidazole (MeIm) by a facile one-pot reaction in methanol at room temperature. On the basis of the experimental results and the theoretical calculations, the MOF 1 contains paddle–wheel [Co 2 (-CO 2 ) 4 ] secondary building units (SBUs) axially coordinated by a MeIm ligand, which is very beneficial to the electron transfer between the Co(II) ions and oxygen. Combining the photosensitizers ZnTCPP and the electroactive [Co 2 (-CO 2 ) 4 ] SBUs, the 2D MOF 1 possesses an excellent ECL performance, and can be used as a novel ECL probe for rapid nonamplified detection of the RdRp gene of SARS-CoV-2 with an extremely low limit of detection (≈30 aM).

研究领域: 材料科学 · 分析化学 · 病毒学
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TALANTA · 2022 · IF 6.556

Hybridization chain reaction circuit-based electrochemiluminescent biosensor for SARS-cov-2 RdRp gene assay

Kai Zhang,Zhenqiang Fan,Yue Huang,Yuedi Ding,Minhao Xie,Minghe Wang

In this work, we designed an ECL ratiometric biosensor with a three-stranded Y-type DNA (Y-DNA) probe and induced a hybridization chain reaction (HCR) for the highly sensitive detection of SARS-CoV-2 nucleic acid. The important component of this system is the self-assembled Y-Shaped probe based on three nucleic acids. Y1, Y2, and Y3 can be linked by complementary base pairing to Hairpin1 (H1), Hairpin2 (H2), and Ru modified DNA (Ru1), respectively. H1 and H2 can trigger the HCR reaction when activated by the SARS-CoV-2 RdRp gene and the 5′ end of Ru1. The 5′ end of Ru1 is modified with the Ru complex, which can produce a strong electrochemiluminescence luminescence signal at 620 nm under an applied voltage. Through the amplification of Y-DNA-induced HCR reaction, Ru1 on the electrode surface gradually increased, the ECL signal at 460 nm was gradually quenched, and the signal at 620 nm was steadily generated. The SARS-CoV-2 RdRp gene can be quantified according to the degree of decrease of ECL signal at 460 nm and the increase of ECL signal at 620 nm. Combining the two signal amplification strategies, this ratiometric ECL biosensor can accurately and efficiently detect the target gene with a detection limit of 59 aM.

研究领域: 生物医学工程 · 分析化学 · 分子诊断学 · 病毒学
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JOURNAL OF HAZARDOUS MATERIALS · 2023 · IF 13.6

Entropy-driven assisted T7 RNA polymerase amplification-activated CRISPR/Cas13a activity for SARS-CoV-2 detection in human pharyngeal swabs and environment by an electrochemiluminescence biosensor

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 · 分子生物学 · 病毒学 · 诊断技术 · 纳米技术
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ACS Applied Materials & Interfaces · 2021 · IF 9.229

Rational Engineering of the DNA Walker Amplification Strategy by Using a Au@Ti3C2@PEI-Ru(dcbpy)32+ Nanocomposite Biosensor for Detection of the SARS-CoV-2 RdRp Gene

Bo Yao,Jing Zhang,Zhenqiang Fan,Yuedi Ding,Bin Zhou,Runlin Yang,Jianfeng Zhao,Kai Zhang

The detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial for preventing and controlling infectious diseases and disease treatment. In this work, a Au@Ti3C2@PEI-Ru(dcbpy)32+ nanocomposite-based electrochemiluminescence (ECL) biosensor was rationally designed, which realized sensitive detection of the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2. In addition, a DNA walker was also used to excise the hairpin DNAs under the action of Nb.BbvCI endonuclease. Furthermore, model DNA–Ag nanoclusters (model DNA–AgNCs) were used to quench the initial ECL signal. As a result, the ECL biosensor was used to sensitively detect the SARS-CoV-2 RdRp gene with a detection range of 1 fM to 100 pM and a limit of detection of 0.21 fM. It was indicated that the ECL biosensor had a great application potential for clinical medical detection. Furthermore, the DNA walker amplification also played a reliable candidate strategy for other detection methods.

研究领域: 生物传感器 · 纳米技术 · 病毒学 · 分析化学
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CHEMICAL ENGINEERING JOURNAL · 2021 · IF 13.273

A pH-engineering regenerative DNA tetrahedron ECL biosensor for the assay of SARS-CoV-2 RdRp gene based on CRISPR/Cas12a trans-activity

Kai Zhang,Zhenqiang Fan,Yuedi Ding,Minhao Xie

In this work, we constructed an exonuclease III cleavage reaction-based isothermal amplification of nucleic acids with CRISPR/Cas12a-mediated pH-induced regenerative Electrochemiluminescence (ECL) biosensor for ultrasensitive and specific detection of SARS-CoV-2 nucleic acids for SARS-CoV-2 diagnosis. The triple-stranded nucleic acid in this biosensor has an extreme dependence on pH, which makes our constructed biosensor reproducible. This is essential for effective large-scale screening of SARS-CoV-2 in areas where resources are currently relatively scarce. Using this pH-induced regenerative biosensor, we detected the SARS-CoV-2 RdRp gene with a detection limit of 43.70 aM. In addition, the detection system has good stability and reproducibility, and we expect that this method may provide a potential platform for the diagnosis of COVID-19.

研究领域: 生物技术 · 纳米技术 · 分子生物学 · 分析化学 · 病毒学
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MICROCHEMICAL JOURNAL · 2023 · IF 4.8

An electrochemiluminescence sensing platform combining entropy-driven amplification strategy and inverted DNA tetrahedral nanomachines for detecting SARS-CoV-2 specific gene

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 · 纳米技术 · 分析化学 · 分子生物学 · 病毒学
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