ANALYTICA CHIMICA ACTA
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2019
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IF 5.256
Yongkang Ye,Wuwen Yan,Yaqian Liu,Shudong He,Xiaodong Cao,Xuan Xu,Haisong Zheng,Sundaram Gunasekaran
A rapid and sensitive electrochemical biosensor was constructed to detect Salmonella using invA gene biosensor. The biosensing was based on polyrrole-reduced graphene oxide (PPy-rGO) nanocomposite modified glassy carbon electrode (GCE) and signal amplification with horseradish peroxidase-streptavidin biofunctionalized gold nanoparticles (AuNPs-HRP-SA). PPy-rGO was prepared at 60 °C by chemical reduction of PPy-functionalized graphene oxide (PPy-GO) that was synthesized by in situ polymerization at room temperature . The detection signal was amplified via enzymatic reduction of H 2 O 2 in the presence of hydroquinone (HQ) using AuNPs-HRP-SA as nanotag. Under optimal conditions, the differential pulse voltametric (DPV) signal from the biosensor was linearly related to the logarithm of target invA gene concentrations from 1.0 × 10 −16 to 1.0 × 10 −10 M, and the limit of detection (LOD) was 4.7 × 10 −17 M. The biosensor can also detect Salmonella in the range of 9.6 to 9.6 × 10 4 CFU mL −1 , with LOD of 8.07 CFU mL −1 . The biosensor showed good regeneration ability , acceptable selectivity , repeatability and stability, which bode well as an alternative method for Salmonella screening.
研究领域:
生物技术
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食品科学
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微生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2023
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IF 7.4
Liping Xia,Juxin Yin,Jianjian Zhuang,Weihong Yin,Zheyu Zou,Ying Mu
Rapid and sensitive pathogen detection methods are critical for disease diagnosis and treatment. RPA-CRISPR/Cas12 systems have displayed remarkable potential in pathogen detection. A self-priming digital PCR chip is a powerful and attractive tool for nucleic detection. However, the application of the RPA-CRISPR/Cas12 system to the self-priming chip still has great challenges due to the problems of protein adsorption and two-step detection mode of RPA-CRISPR/Cas12. In this study, an adsorption-free self-priming digital chip was developed and a direct digital dual-crRNAs (3D) assay was established based on the chip for ultrasensitive detection of pathogens. This 3D assay combined the advantages of rapid amplification of RPA, specific cleavage of Cas12a, accurate quantification of digital PCR, and point-of-care testing (POCT) of microfluidics, enabling accurate and reliable digital absolute quantification of Salmonella in POCT. Our method can provide a good linear relationship of Salmonella detection in the range from 2.58 × 101 to 2.58 × 104 cells/mL with a limit of detection ∼0.2 cells/mL within 30 min in a digital chip by targeting the invA gene of Salmonella. Moreover, the assay could directly detect Salmonella in milk without nucleic acid extraction. Therefore, the 3D assay has the significant potential to provide accurate and rapid pathogen detection in POCT. This study provides a powerful nucleic detection platform and facilitates the application of CRISPR/Cas-assisted detection and microfluidic chips.
研究领域:
分子生物学
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生物技术
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微生物学
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分析化学
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即时诊断
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2018
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IF 5.667
Yongkang Ye,Yaqian Liu,Shudong He,Xuan Xu,Xiaodong Cao,Yingwang Ye,Haisong Zheng
This work described a sensitive electrochemical sensing method for detection of invA gene sequence of Salmonella by integrating direct signal inducer of silver nanoclusters (AgNCs) and cystine-rich (C-rich) signal probe DNA assembled gold nanoparticls (sDNA-AuNPs). Thiolated capture probe was covalently immobilized onto the glassy carbon electrode, and further hybridized with target DNA sequence (tDNA) and sDNA-AuNPs onto form a sandwich-type structure. The C-rich sDNA in the structure was then used as template for the formation of AgNCs, which can produce the direct voltametric signal during the detection of tDNA. Under optimal conditions, the fabricated biosensor showed good linearity from 1 fM to 0.1 nM with a detection limit of 0.162 fM ( S/N = 3). It exhibited good selectivity, stability and reproducibility and can be applied to detect Salmonella effectively by detecting the polymerase chain reaction products of invA gene.
研究领域:
生物传感器
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电化学
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纳米技术
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微生物学
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分子生物学
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
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2023
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IF 6.2
Yongkang Ye,Wuwen Yan,Tingting Wang,Chenlu Zhang,Kaicheng Wang,Yuexi Lu,Haisong Zheng,Yunlai Tao,Xiaodong Cao,Shudong He,Yunfei Li
Simultaneous detection of multiple foodborne pathogens is of great importance for ensuring food safety. Herein, we present a sensitive dual-channel electrochemical biosensor based on copper metal organic frameworks (CuMOF) and lead metal organic framework (PbMOF) for simultaneous detection of Salmonella typhimurium ( S. typhimurium ) and Listeria monocytogenes ( L. monocytogenes ). The MOF-based nanotags were prepared by functionalizing gold nanoparticles loaded CuMOF (Au@CuMOF) and PbMOF (Au@PbMOF) with signal DNA sequences 1 (sDNA1) and sDNA2, respectively. By selecting invA of S. typhimurium and inlA gene of L. monocytogenes as targe sequences, a sandwich-typed dual-channel biosensor was developed on glassy carbon electrodes (GCE) through hybridization reactions. The sensitive detection of S. typhimurium and L. monocytogenes was achieved by the direct differential pulse voltametric (DPV) signals of Cu 2+ and Pb 2+ . Under optimal conditions, channel 1 of the biosensor showed linear range for invA gene of S. typhimurium in 1 × 10 −14 −1 × 10 −8 M with low detection limit (LOD) of 3.42 × 10 −16 M ( S/N = 3), and channel 2 of the biosensor showed linear range for inlA gene of L. monocytogenes in 1 × 10 −13 −1 × 10 −8 M with LOD of 6.11 × 10 −15 M (S/N = 3). The dual-channel biosensor showed good selectivity which were used to detect S. typhimurium with linear range of 5−1.0 × 10 4 CFU mL −1 (LOD of 2.33 CFU mL −1 ), and L. monocytogenes with linear range of 10 − 1.0 × 10 4 CFU mL −1 (LOD of 6.61 CFU mL −1 ).
研究领域:
食品安全
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生物传感器
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纳米技术
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电化学
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病原体检测
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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）