Early antibiotic resistance determinants (ARDs) detection in humans or animals is crucial to counteract their propagation. The ARDs quantification is fundamental to understand the perturbation caused by disruptors, such as antibiotics, during therapies. Forty-three qPCRs on the most diffused ARDs and integrons among human and animal Enterobacterales, and one on the 16S rDNA for bacteria quantification, were developed. The qPCRs, using hydrolysis probes, operated with a unique amplification condition and were tested analytically and diagnostically performing 435 reactions on five positive and negative controls for each qPCR. Diagnostic sensitivity and specificity were confirmed by PCR and genome sequencing of control isolates, demonstrating 100% performance for all qPCRs. An easy and rapid discrimination method for the epidemiologically relevant bla is provided. This large, noncommercial qPCRs inventory could serve for precise quantification of ARDs, but also as a rapid screening tool for surveillance purposes, providing the basis for further high-throughput developments.
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http://dx.doi.org/10.1016/j.diagmicrobio.2021.115328 | DOI Listing |
Biosensors (Basel)
January 2025
Furong Labratory, Changsha 410083, China.
A fluorescence probe for "switch-on" detection of alkaline phosphatase (ALP) was developed based on Au nanoclusters anchored MnO nanosheets (Au NCs-MnO NSs), which were synthesized using bovine serum albumin (BSA) as template through a simple one-pot approach. In the sensing system, MnO NSs function as both energy acceptors and target identifiers, effectively quenches the fluorescence of Au NCs via fluorescence resonance energy transfer (FRET). The presence of ALP catalyzes the hydrolysis of L-ascorbic acid-2-phosphate (AAP) to ascorbic acid (AA), reducing MnO NSs to Mn and facilitate the fluorescence recovery of Au NCs.
View Article and Find Full Text PDFChem Sci
January 2025
Key Laboratory for Advanced Materials, Institute of Fine Chemicals, Feringa Nobel Prize Scientist Joint Research Center, Frontiers Science Center for Materiobiology and Dynamic Chemistry, School of Chemistry and Molecular Engineering, Center of Photosensitive Chemicals Engineering, East China University of Science and Technology Shanghai 200237 China
Given that proteins with minor variations in amino acid sequences cause distinct functional outcomes, identifying and quantifying similar proteins is crucial, but remains a long-standing challenge. Herein, we present a two-dimensional orthogonal fluorescence and chemiluminescence design strategy for the probe DCM-SA, which is sequentially activated by albumin-mediated hydrolysis, exhibiting light-up fluorescence and photo-induced cycloaddition generating chemiluminescence, enabling orthogonal signal amplification for discrimination of subtle differences between similar proteins. By orthogonalizing these dual-mode signals, a two-dimensional work curve of fluorescence and chemiluminescence is established to distinguish and quantify similar proteins HSA and BSA.
View Article and Find Full Text PDFAnal Chim Acta
February 2025
Shenzhen Key Laboratory of Synthetic Genomics, Guangdong Provincial Key Laboratory of Synthetic Genomics, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China. Electronic address:
Background: High-precision and broad-range pH detection is critical for health status assessment, such as signal transduction, enzyme activity, endocytosis, and cell proliferation and apoptosis. Although pH-responsive ratiometric fluorescent probes offer an effective pH monitoring strategy, their preparation often requires multi-step modification and decreases fluorescence efficiency and stability. Herein, we developed a simple method to prepare fluorescent Si dots with dual emission centers for high-precision and broad-range pH monitoring, and the detection of urease based on pH-responsive Si dots and pH monitoring in living cell was further explored.
View Article and Find Full Text PDFTalanta
January 2025
The Higher Educational Key Laboratory for Nano Biomedical Technology of Fujian Province, Department of Pharmaceutical Analysis, School of Pharmacy, Fujian Medical University, Fuzhou, 350122, China. Electronic address:
Electrochemical bioassays that rely on sensor interfaces based on immobilized DNA probes often encounter challenges such as complex fabrication processes and limited binding efficiency. In this study, we developed a novel electrochemical bioassay that bypasses the need for probe immobilization by employing a solution-phase nucleic acid reaction to create interfacial barriers on unmodified electrodes, enabling rapid, just-in-time sensor interface formation. Specifically, a 3'-phosphorylated recognition probe was used to identify the target microRNA-21 (miR-21), followed by target recycling facilitated by duplex-specific nuclease (DSN), which resulted in extensive hydrolysis of the recognition probe into DNA fragments with 3'-hydroxyl ends.
View Article and Find Full Text PDFJ Am Chem Soc
January 2025
Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Site-selective cleavage of the peptide backbone in proteins is an important class of post-translational modification (PTM) in nature. However, the organic chemistry for such site-selective peptide bond cleavages has yet to be fully explored. Herein, we report cysteine -formylation as a means of selective protein backbone cleavage.
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