The sensitive detection and quantification of DNA targets in the food industry and in environmental and clinical settings are issues of utmost importance in ensuring contamination-free food, monitoring the environment, and battling disease. Selective probes coupled with powerful amplification techniques are therefore of major interest. In this study, we set out to create an integrated microchemical chip that benefits from microfluidic chip technology in terms of sensitivity and a strong detection methodology provided jointly by padlock probes and rolling circle amplification (RCA). Here, we have integrated padlock probes and RCA into a microchip. The chip uses solid phase capture in a microchannel to enable washing cycles and decrease analytical area, and employs on-bead RCA for single-molecule amplification and detection. We investigated the effects of reagent concentration and amount of padlock probes, and demonstrated the feasibility of detecting Salmonella.
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http://dx.doi.org/10.1039/b927460j | DOI Listing |
Water Res
January 2025
Department of Aquatic Sciences and Assessment, Swedish University of Agricultural Sciences, Box 7050, 75007, Uppsala, Sweden. Electronic address:
Wastewater-based surveillance (WBS) allows the analysis of pathogens, chemicals or other biomarkers in wastewater to derive unbiased epidemiological information at population scale. After re-gaining attention during the SARS-CoV-2 pandemic, the field holds promise as a surveillance and early warning system by tracking emerging pathogens with pandemic potential. Expanding the current toolbox of analytical techniques for wastewater analysis, we explored the use of Hyperplex PCR (hpPCR) to analyse SARS-CoV-2 mutations in wastewater samples collected weekly in up to 22 sites across Sweden between October 2022 and December 2023.
View Article and Find Full Text PDFACS Omega
December 2024
Department of Gastroenterology, Xiamen University Affiliated Chenggong Hospital, Xiamen City, Fujian Province 361003, China.
MicroRNAs (miRNAs), which play critical roles in regulating gene expression and cell functions, are recognized as potential biomarkers for various human diseases, including gastric ulcers. The reliable, specific, and sensitive detection of miRNA is highly recommended for the clinical diagnosis and therapy of different diseases. Herein, we depict a label-free and low-background fluorescent assay for the highly sensitive detection of miRNAs by coupling target miRNA-triggered cyclization of a padlock, circular padlock-mediated catalytic hairpin assembly (CHA), and primer exchange reaction (PER)-assisted signal generation.
View Article and Find Full Text PDFBiosens Bioelectron
March 2025
State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha, 410082, PR China.
Accurate and sensitive detection of Pax-5a gene is the basis of early diagnosis and prediction of acute leukemia. This research aims to develop a universal dual-mode sensing method enables ultrasensitive gene detection based on smart control of DNA amplification by nucleic acid beacons e to form programmed dendrimer. The Pax-5a target gene triggers the opening of smart gate hairpin probe (Hp), exposing the stem sequence as the primer to bind with padlock probe for rolling circle amplification (RCA).
View Article and Find Full Text PDFAnal Methods
January 2025
Department of Colorectal Surgery, College of Clinical Medicine for Oncology, Fujian Medical University, Fuzhou, Fujian, China.
MicroRNA (miRNA) is a promising biomarker for the early diagnosis of pancreatic cancer. To enable sensitive and reliable miRNA detection, we have developed a one-pot isothermal CRISPR/Dx detection system by combining rolling circle amplification (RCA) and CRISPR/Cas12a. RCA and CRISPR/Cas12a reactions are carried out in a single closed tube, bypassing the transferring step.
View Article and Find Full Text PDFAdv Neurobiol
November 2024
Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
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