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Guide DNA dephosphorylation-modulated Pyrococcus furiosus Argonaute fluorescence biosensor for the detection of alkaline phosphatase and aflatoxins B1.
Huang, Zhengzhang; Wei, Luyu; Zhou, Yanan; Li, Yingjun; Chen, Yiping.
Afiliación
  • Huang Z; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
  • Wei L; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
  • Zhou Y; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
  • Li Y; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China. Electronic address: yingjun@mail.hzau.edu.cn.
  • Chen Y; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Academy of Food Interdisciplinary Science, School of Food Science and Technology, Dalian Polytechnic University, Dalian, 116034, Liaoning, China; State Key Laboratory of Marine Food Processing and
Biosens Bioelectron ; 265: 116692, 2024 Dec 01.
Article en En | MEDLINE | ID: mdl-39208510
ABSTRACT
Foodborne hazardous factors pose a significant risk to public health, emphasizing the need for the development of sensitive and user-friendly detection strategies to effectively manage and control these risks in the food supply chain. Pyrococcus furiosus argonaute (PfAgo)-based biosensing approaches have been extensively explored due to its built-in signal amplification. However, the property that PfAgo is a DNA-guided DNA endonuclease has enabled almost all the existing PfAgo-based reports to be used for the detection of nucleic acids. To lend PfAgo toolbox to extended non-nucleic acid detection, we systematically investigated the mechanism characteristic of PfAgo' preference for guide DNA (gDNA) and proposed a gDNA dephosphorylation-modulated PfAgo sensor for the detection of non-nucleic acid targets. Our results indicated that PfAgo exhibits preference for 5'-phosphorylated gDNA at a specific ratio of PfAgo to gDNA concentration. Leveraging this PfAgo' preference and the dephosphorylation activity of alkaline phosphatase (ALP), ALP could be detected as low as 2.7 U/L. Furthermore, the PfAgo was coupled with immunolabelled ALP to develop a PfAgo-based fluorescence immunosensor, which achieves aflatoxins B1 detection with a detection limit of 29.89 pg/mL and exhibits satisfactory recoveries in wheat and maize samples. The developed method broadens the application scope of PfAgo toolbox, and provides a simple, sensitive, and universal detection platform for a variety targets.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Aflatoxina B1 / Pyrococcus furiosus / Fosfatasa Alcalina Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Aflatoxina B1 / Pyrococcus furiosus / Fosfatasa Alcalina Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido