Nucleic acid tests integrated into digital point-of-care (POC) diagnostic systems have great potential for the future of health care. However, current methods of DNA amplification and detection require bulky and expensive equipment, many steps, and long process times, which complicate their integration into POC devices. We have combined an isothermal DNA amplification method, recombinase polymerase amplification, with an electrochemical stem-loop (S-L) probe DNA detection technique. By combining these methods, we have created a system that is able to specifically amplify and detect as few as 10 copies/μL DNA with a total time to result of 70-75 min.
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http://dx.doi.org/10.1021/acsomega.0c00341 | DOI Listing |
Int J Mol Sci
December 2024
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Genetically modified (GM) herbicide-tolerant soybean 'Zhonghuang 6106', which introduces a glyphosate-resistant gene, ensures soybean yield while allowing farmers to reduce the use of other herbicides, thereby reducing weed management costs. To protect consumer rights and facilitate government supervision, we have established a simple and rapid on-site nucleic acid detection method for GM soybean 'Zhonghuang 6106'. This method leverages the isothermal amplification characteristics of RPA technology and the high specificity of CRISPR-Cas12a to achieve high sensitivity and accuracy in detecting GM soybean components.
View Article and Find Full Text PDFAnimals (Basel)
December 2024
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
Flamingos () are among the oldest birds worldwide and are loved by people for their bright red feathers. In addition, flamingos are sexually monomorphic birds, and distinguishing between males and females is difficult. The polymerase chain reaction (PCR) is widely used for sex identification.
View Article and Find Full Text PDFJ Agric Food Chem
January 2025
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
() is a major pathogenic bacterium responsible for bacterial foodborne diseases, making its rapid, specific, and accurate detection crucial. In this study, we develop a ratiometric biosensor based on the recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats/CRISPR associated protein 12a (RPA-CRISPR/Cas12a) system and Eu-metal-organic framework (Eu-MOF) fluorescent nanomaterials for the high-sensitivity detection of , combining with RPA for efficient isothermal amplification, this sensor enhances specificity and sensitivity by utilizing the target activation of CRISPR/Cas12a. The Eu-MOF serves a dual function, providing stable red fluorescence as a reference signal and adsorbing FAM-labeled probes for fluorescence quenching, forming a dual-signal system that significantly reduces background interference.
View Article and Find Full Text PDFAnal Chim Acta
January 2025
Department of Quality Control Material R&D, Shanghai Center for Clinical Laboratory, Shanghai, PR China; Department of Molecular Biology, Shanghai Center for Clinical Laboratory, Shanghai, PR China; Department of Molecular Diagnostic Innovation Technology, Shanghai Academy of Experimental Medicine, Shanghai, PR China. Electronic address:
Background: The current pandemic of 2022 global mpox (formerly known as monkeypox), caused by infection with monkeypox virus (MPXV), has now reached over 120 countries. This constitutes a critical public health issue requiring effective disease management and surveillance. Rapid and reliable diagnosis is conducive to the control of infection, early intervention, and timely treatment.
View Article and Find Full Text PDFInt J Biol Macromol
January 2025
NHC Key Laboratory of Tropical Disease Control, School of Tropical Medicine & The Second Affiliated Hospital, Hainan Medical University, Haikou 571199, PR China. Electronic address:
Nucleic acids detection is essential for diagnosing pathogens; however, traditional methods usually face challenges such as low sensitivity, lengthy reaction times, and strict temperature requirements. This study develops a novel photoelectrochemical (PEC) biosensor that integrates recombinase polymerase amplification (RPA) with a 3D-array titania (TiO) nanorods nanorod electrode, addressing the challenge of achieving sensitive detection of RPA-amplified nucleic acids products, thereby enabling earlier and more reliable pathogen detection. The biosensor utilizes a triple-binding mode involving FITC antibodies, target nucleic acids, and an HRP-streptavidin sandwich structure, significantly improving the bio-functionalization of the electrode surface.
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