Frontiers in Cellular and Infection Microbiology
·
2025
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IF 4.8
Yizhuo Zhang,Luqin Lv,Su Xu,Yijian Chen
Introduction: Clostridioides difficile (C. difficile), a human pathogen that causes diarrhea and colon lesions, has garnered widespread attention. Rapid and accurate detection of bacterial virulence factors is essential for the diagnosis of C. difficile infection (CDI). To date, numerous laboratory tests have been developed; however, none fully meet the combined requirements of speed, cost-effectiveness, portability, sensitivity, and specificity. Molecular diagnostic technologies based on CRISPR-Cas systems have provided a promising solution to this challenge. Nonetheless, the limited compatibility between pre-amplification and CRISPR cleavage, coupled with the inherent selectivity of CRISPR systems for protospacer adjacent motif (PAM) sequences near the target site, poses additional constraints on the broader adoption of this approach.Methods: Here, we developed PAM-unconventional, One-step LAMP/CRISPR-Cas12b (POLC) detection platforms for the toxin-encoding genes tcdA and tcdB of C. difficile.Results: The POLC platforms operated at 60 °C, enabling result interpretation either through fluorescence intensity measurements or direct visualization under UV light. The limits of detection (LoDs) ranged from 3 to 14 copies/μL using a fluorescence reader and from 6 to 18 copies/μL via direct observation. Compared to qPCR, which typically requires over an hour, the POLC platforms reduced the detection time to approximately 40 minutes. Each reaction cost approximately USD 6.5, offering a substantial cost saving compared to qPCR-based commercial kits (over USD 10 per test). In clinical validation with 55 fecal samples, the tcdA POLC assay achieved 86.4% sensitivity and 84.8% specificity, while the tcdB POLC assay demonstrated 96.6% sensitivity and 100% specificity, using qPCR as the reference standard.Discussion: Our research presents innovative CRISPR-based one-step nucleic acid detection platforms that eliminate canonical PAM sequence requirements. These platforms exhibit high sensitivity and specificity while achieving rapid detection under simple conditions, making them promising candidates for clinical diagnostics and point-of-care testing (POCT).
研究领域:
医学诊断
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2024
·
IF 12.2
Le Liang,Su-qian Cai,Yan-li Leng,Chan Huang,Yong-qing Liu,Ye Wang,Li Luo,Mei Han,Xiao-hong Li,Xiao-hua Cai
Ciprofloxacin (CIP) and gallium (Ga) are widely used in many fields and their excessive presences will pose serious threats to the environment and life health. Therefore, developing an intelligent detection method for rapidly tracking and determination of CIP and Ga 3 ⁺ concentration in environments and organisms is of great significance. In this work, a europium-doped carbon dots (Eu-CDs) with unique structure and performances was prepared by a one-pot hydrothermal method. Eu-CDs can efficiently overcome the problem of aggregation-induced fluorescence quenching in complex environments to track CIP and Ga 3⁺ with highly sensitivity. The "antenna effect" of non-radiative energy transfer is introduced to explain the pathway on fluorescence transition of Eu-CDs through DFT theoretical calculations and mechanism investigations, which provide a theoretical basis for the design and development of novel carbon dots with specific functions. Eu-CDs can successfully be applied to fluorescence imaging in organisms, moreover, it also exhibits excellent application potentials in the field of anti-tumor and antibacterial. In addition, a portable intelligent detection platform based on Eu-CDs-hydrogel device have been developed, which provides a new strategy for rapidly detection of CIP and Ga 3⁺ in real samples.
研究领域:
分析化学
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环境科学
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生物医学工程
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY
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2022
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IF 2.731
Xuechun Yan,Mingbin Liu,Hongbo Li,Suqin Wang
Silver is widely used, but the pollution caused by silver ions is also a problem that cannot be ignored. Detection of heavy metal ions based on bulky and costly analytical instruments had been developed very maturely in the laboratory. At present, the main detection methods of silver ion concentration include inductively coupled plasma mass spectrometry, atomic absorption spectrometry and inductively coupled plasma/atomic emission spectrometry. However, these testing methods have many problems to be solved, such as the high of instruments, high operating cost, numerous operational requirements, long testing time and complex processing technology. Nowadays, with the rapid and powerful development of smartphone technology, the new analytical methods based on smartphones equipped with a camera are being carried in the certain applied ideas. Here, the paper introduces a hand-held detection platform smartphone-based that allows the quantification of silver(I) ions with the detection limit of 0.659 mM. The Colour Grab software of android smartphone was applied to obtain R, G, B values of the photo of Rhodamine B solution remaining fluorescence, and then calculated the corresponding grey values by grey formula. The detection platform based on smart phones has the characteristics of cost-effective design and simple operation, as well as high sensitivity and selectivity. Therefore, compared with other detection methods, this method can be widely and conveniently applied to the detection of silver ions in the environment.
研究领域:
环境科学
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分析化学
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产品关联
DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2025
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IF 11.3
Wenjing Wang,Rui Feng,Kaiyue Wei,Jiashuo Xu,Wenhui Dong,Jiao Li,Jianan Sun,Sai Wang,Xiangzhao Mao
Excessive histamine in spoiled seafood poses considerable health hazards to consumers, yet its detection is challenging due to complicated sample preparation and detection methodologies. Herein, an integrated colorimetric platform containing Poly (vinyl alcohol) (PVA)/ hyaluronic acid (HA) microneedle patchs-assisted extraction and aptasensor-based detection was reported. The developed PVA/HA microneedle patches facilitated on-site histamine extraction from seafood through a two-minute press-and-peel procedure. To enhance detection efficacy, strategies for generating high-affinity aptamers with specific terminal-fixed structures and constructing AuNPs@FeP-chitosan oligosaccharide (COS) nanozyme boosting catalytic efficiency were proposed. Utilizing the aptamer HIS3-T2 in conjunction with the nanozyme, a colorimetric aptasensor was developed. Integrated with the patches, the aptasensor achieved high sensitivity and selectivity, detecting histamine within a range of 2~800 nM with a limit of detection (LOD) of 1.89 nM. Validated on real-world salmon and shrimp samples, this integrated system promises rapid and accurate histamine monitoring, offering great reference for similar applications in food quality control.
研究领域:
生物传感技术
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食品安全
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分析化学
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产品关联
DOI
PubMed
高影响因子
TALANTA
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2025
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IF 6.1
Hao Gong,Yang Li,Yaru Liu,Yuting Shan,Yijie Wang,Cuiping Ma,Chao Shi
Pathogenic bacteria represent a significant threat to public health and the global economy due to their high infectiousness and lethality. Consequently, the rapid and accurate identification of these bacteria remains a major challenge. Electrochemical amplification-free detection methods have shown several advantages, including rapidity, simplicity and high sensitivity. However, these methods also possess certain drawbacks, such as the inefficient solid-phase hybridization of long-chain nucleic acids on the electrode surface, it is challenging to directly detect long-chain nucleic acids. In response, we have established a novel sensor platform for efficient and rapid detection of long chain nucleic acids based on the principle of HRP signal amplification. This platform incorporates macromolecular crowding agents and electric field assistance to enhance pathogenic bacteria detection efficiency in clinical samples without amplification. The detection process begins by introducing the target into a strand displacement reaction system containing a crowding agent. The crowding effect of the crowding agent is such that the long chain target rapidly replaces the biotinylated probe. Subsequently, an electric field is applied to the surface of the screen-printed electrodes for rapid hybridization, which has been modified with a capture probe. This assay is straightforward and expeditious, reducing hybridization time from 150 min to 90 s. The sensor platform showed a detection linear range of 10 2 –10 7 CFU mL −1 with the limit of detection of 10 CFU mL −1 within 25min. This platform showcases the significant potential for on-site detection of pathogenic bacteria in clinical settings and is expected to be integrated into existing diagnostic equipment.
研究领域:
生物传感
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医学诊断
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2020
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IF 3.209
Shuyu Mei,Bingru Liu,Xiaoli Xiong,Xu Hun
PtCuCo trimetallic alloys (PtCuCo-TAs) are synthesized by one-step reduction. The chemiluminescence (CL) properties of PtCuCo-TAs are studied systemically. PtCuCo-TAs show good catalyzing for luminol-H 2 O 2 system. A CL platform is developed for the detection of miRNA-21 using PtCuCo-TAs as nanozyme catalyzer. In the CL detection platform, H1 (Hairpin DNA1) is immobilized onto magnetic beads (MBs) firstly. In the presence of miRNA-21, H1 is opened. H2 (Hairpin DNA2) then hybridizes with H1. Meanwhile, a “cleat” in the end of miRNA-21 with a fewer bases complementary is formed to prevent miRNA-21 dissociating from H1. This miRNA-21 hybridizes to another H1. When cpDNA-PtCuCo-TAs which consisted with cDNA (Complementary strand of probe DNA) and pDNA-PtCuCo-TAs (PtCuCo-TAs labeled with probe DNA) are added, the ssDNA region of H1 reacts with the toehold domain of probe DNA and cDNA is released resulting pDNA-PtCuCo-TAs being captured. With this process repeatedly, a lot of pDNA-PtCuCo-TAs are captured onto MBs. After separation and washing, the precipitate and H 2 O 2 are put into the 96-well and luminol solution is injected. The CL signal is produced by PtCuCo-TAs catalyzing luminol-H 2 O 2 system. The amount of miRNA-21 is detected with CL signal. This CL platform performs with limit of detection 0.167 fM and has good selectivity over other RNA.
研究领域:
生物化学
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分析化学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
LANGMUIR
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2018
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IF 3.789
Lele Zhou,Qiongzheng Hu,Qi Kang,Ming Fang,Li Yu
The detection of l-phenylalanine (l-Phe) has become one of the most pressing issues concerning diagnosis and treatment of phenylketonuria in neonates; however, a simple and robust methodology is yet to be developed. Here, the application of novel liquid crystals (LCs)-sensing platform for sensitive, selective, and label-free detection of l-Phe was reported at the first time. We devised a strategy to fabricate the sodium monododecyl phosphate (SMP)-decorated LC sensing platform with the appearance of dark. Then, a dark to bright (D–B) optical images alteration of LCs was observed after transferring alkaline phosphatase (ALP) to the interface, owing to cleavage of SMP induced by ALP. LCs remained dark images after the SMP-decorated interface in contact with the pre-incubated ALP and l-Phe. Such optical appearance resulted from the inhibition of ALP by l-Phe, which was further verified by the isothermal titration calorimetry (ITC). The strategy was applied to sensing l-Phe, which have been proven to allow for sensitively and selectively differentiation of l-Phe from interfering compounds with similar aromatic groups, as well as seven other essential amino acids. More importantly, the detection limit of l-Phe reached 1 pg/mL in urine samples, further demonstrating its value in the practical applications. Results obtained in this study clearly demonstrated the superiority of LCs toward the l-Phe detection, which can pave a way for the development of high performance and robust probes for l-Phe detection in clinical applications.
研究领域:
生物化学
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医学诊断
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材料科学
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产品关联
DOI
PubMed
高影响因子
LUMINESCENCE
·
2024
·Vol. 39
·期 2
·e4689
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IF 3.2
Mingyu Gan,Yingqi Wang,Fei Wang,Jie Tan,Yuheng Pei,Jianhua Wang,Martin M. F. Choi,Wei Bian
A new type of polyethyleneimine-protected copper nanoclusters (PEI-CuNCs) is favorably developed by a one-pot method under mild conditions. The obtained PEI-CuNCs is characterized by X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, Fourier-transform infrared (FTIR) spectroscopy and other techniques. It is worth noting that the proposed PEI-CuNCs demonstrate a selective response to chromium(VI) over other competitive species. Fluorescence quenching of PEI-CuNCs is determined to be chromium(VI) concentrations dependence with a low limit of detection of 8.9 nM. What is more, the as-developed PEI-CuNCs is further employed in building a detection platform for portable recognition of chromium(VI) in real samples with good accuracy. These findings may offer a distinctive strategy for the development of methods for analyzing and monitoring chromium(VI) and expand their application in real sample monitoring.
研究领域:
分析化学
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环境科学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
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IF 12.6
Hua Wang,Yang Sun,Yuan Zhou,Yujie Liu,Shuo Chen,Wenbo Sun,Zidong Zhang,Junqing Guo,Cheng Yang,Zhengping Li,Lei Chen
At present, the 100% case fatality and the cross-infection of virus strains make the ASFV 's harm to society continue to expand. The absence of an effective commercial vaccine poses early detection remains the most effective means of curbing ASFV infection. Here, we report a cascaded detection platform based on the CRISPR-Cas12a system combined with graphene field-effect transistor sensors. The cascade platform could detect ASFV as low as 0.5 aM within 30 min and achieve typing of wild and vaccine strains of ASFV in a single detection system. The evaluation of 16 clinical samples proved that, compared with the gold standard Real-time PCR method, this platform has outstanding advantages in sensitivity, specificity and typing. Combining CRISPR-Cas12a's high specificity with the bipolar electric field effect of graphene field-effect transistor, the cascade platform is expected to achieve clinical application in the field of DNA disease detection, and provides a new direction for multi-strain disease typing.
研究领域:
分子生物学
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病毒学
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生物传感器
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DOI
PubMed
高影响因子
Journal of Materials Chemistry A
·
2015
·Vol. 3
·期 38
·19455-19460
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IF 7.443
Zhigang Geng,Haimin Zhang,Qizhong Xiong,Yunxia Zhang,Huijun Zhao,Guozhong Wang
In this work, a three-dimensional (3D) chitosan hydrogel with superior fluorescence properties was successfully fabricated by modifying chitosan fibers with glutaric dialdehyde (GD) via a simple cross-linking approach. The resulting three-dimensional fluorescent chitosan hydrogel (3D-FCH) with hydrophilic properties exhibited a strong blue fluorescence emission at an excitation wavelength of 337 nm. The fluorescence mechanism of the as-synthesized 3D-FCH was investigated and proposed in detail using X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FT-IR) techniques. As a solid-phase fluorescent probe, the 3D-FCH was used to selectively and sensitively determine mercury(II) (Hg2+) ions in aqueous media. The results demonstrated that a prominent fluorescence quenching at 401 nm was observed in the presence of Hg2+ with a linear response range of 5.0–50 nM and an estimated limit of detection of 0.9 nM. The fluorescence quenching mechanism could be ascribed to the strong complexation between Hg2+ and the GD fluorophore with a conjugate structure. Moreover, the porous structure of the chitosan hydrogel and the high adsorption capacity of the chitosan fibers in the hydrogel could be very favorable for the rapid fluorescence determination of Hg2+. This work may pave a new way to develop low-cost fluorescent chitosan hydrogels as solid-phase fluorescence determination platforms to replace traditional liquid-phase fluorophores for application in the fluorescence detection of heavy metal ions.
研究领域:
分析化学
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环境科学
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材料科学
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产品关联
DOI
高影响因子
Animal Nutrition
·
2022
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IF 5.285
Chizhu Ding,Xiang Chen,Xiaoyu Chen,Yue Liu,Menglin Xia,Ziyi He,Qinshu Kang,Xianghua Yan
Lysine is one of the essential amino acids and plays a vital role in the growth, development and health of pigs. Blood lysine concentration is a direct indication of lysine status; however, current methods can not satisfy the demands for rapid and on-site lysine concentration measurement of swine serum. Here, we developed blue-emissive nitrogen-doped carbon dots as a fluorescence probe for the determination of lysine with high fluorescence quantum yield, stability, sensitivity and specificity. The carbon dots were entrapped within hydrogel microstructures to fabricate microfluidic chips for rapid assay for lysine quantification. We further developed an imaging attachment to integrate the microfluidic chip and a smartphone into a portable point-of-care testing platform. This platform requires only 3 μL sample and has a linear detection range of 25 to 300 μmol/L with a limit of detection less than 16 μmol/L, which covers the normal range of lysine concentration in swine serum. We tested lysine concentration in swine serum using this platform with high accuracy, low sample consumption, and within 3 min. Together, these results may provide a rapid and portable platform for dynamic monitoring of swine lysine status and contribute to precise feed formula modulation with low-protein diet strategy.
研究领域:
生物化学
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分析化学
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畜牧学
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DOI
PubMed
高影响因子
Journal of Analysis and Testing
·
2021
·Vol. 6
·期 3
·327-334
·
IF 0
Qi Wen-Jing,He Hong-Kun,Tian Xue,Song Yi,Li Xue-Ni,Li Rong,Hu Ping-Ping,Huang Xiao-Mei
We established a fluorescence “turn-on” sensing platform for glutathione (GSH) detection utilizing chitosan-based glutaraldehyde non-conjugated polymers (GCPF) as nanomaterials via fluorescence resonance energy transfer (FRET) principle. Owing to the overlapping property of absorption spectrum of MnO 2 nanosheets with fluorescence spectrum of GCPF, fluorescent intensity of GCPF was quenched in the presence of MnO 2 nanosheets because of FRET principle. When GSH was added, MnO 2 nanosheets were reduced and decomposed into large amounts of Mn 2+ ions owing to reducing property of GSH. Accordingly, the quenched fluorescent intensity was turned on again. Therefore, this platform based on MnO 2 nanosheet has been first applied to fluorescence “turn-on” detection of GSH from 0.5 to 50 μmol/L with sdetection limit of 84 nmol/L. It showed high selectivity in GSH detection towards other ions and biomolecules such as L-lysine, L-threonine, L-valine, L-glutamic acid and DL-aspartic acid. When it was utilized in detecting GSH in serum samples, satisfactory recoveries ranged from 101.5% to 103.2%, indicating the accuracy of this fluorescence “turn-on” platform in bioanalysis.
研究领域:
生物化学
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分析化学
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DOI
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