Your browser doesn't support javascript.
loading
SATCAS: A CRISPR/Cas13a-based simultaneous amplification and testing platform for one-pot RNA detection and SNPs distinguish in clinical diagnosis.
Wang, Ting; Bai, Linlin; Wang, Guoling; Han, Jingli; Wu, Lixin; Chen, Xuanzhong; Zhang, Hongyu; Feng, Jia; Wang, Yongming; Wang, Rui; Zhang, Xiaohui.
Afiliación
  • Wang T; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China.
  • Bai L; Human Phenome Institute, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, PR China.
  • Wang G; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China.
  • Han J; Peking University People's Hospital, Peking University Institute of Hematology, Beijing, PR China; National Clinical Research Center for Hematologic Disease, Beijing, PR China.
  • Wu L; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China.
  • Chen X; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China.
  • Zhang H; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China. Electronic address: zyiqu@163.com.
  • Feng J; Department of Hematology, Peking University Shenzhen Hospital, Shenzhen Peking University, The Hong Kong University of Science and Technology Medical Center, Shenzhen, 518036, PR China. Electronic address: doray12@hotmail.com.
  • Wang Y; Human Phenome Institute, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, PR China; Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, 200438, PR China. Electronic address: y
  • Wang R; Human Phenome Institute, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, 200438, PR China; Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, 200438, PR China; International Human P
  • Zhang X; Peking University People's Hospital, Peking University Institute of Hematology, Beijing, PR China; National Clinical Research Center for Hematologic Disease, Beijing, PR China. Electronic address: zhangxh@bjmu.edu.cn.
Biosens Bioelectron ; 263: 116636, 2024 Nov 01.
Article en En | MEDLINE | ID: mdl-39116631
ABSTRACT
The clinical diagnosis of pathogen infectious diseases increasingly requires sensitive and rapid RNA detection technologies. The RNA-guided clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system has shown immense potential in molecular diagnostics due to its trans-cleavage activity. However, most Cas13a-based detection methods require an amplicon transcription step, and the multi-step open-tube operations are prone to contamination, limiting their widespread application. Here, we propose an ultrasensitive (single-copy range, ∼aM) and rapid (within 40 min) isothermal one-pot RNA detection platform, termed SATCAS (Simultaneous Amplification and Testing platform based on Cas13a). This method effectively distinguishes viable bacteria (0%-100%) under constant total bacterial conditions, demonstrating its robustness and universality. SATCAS excels in identifying single nucleotide polymorphisms (SNPs), particularly detecting 0.5% drug-resistant mutations. We validated SATCAS by detecting infections in biological samples from 68 HBV, 23 EBV, and 48 SARS-CoV-2 patients, achieving 100% sensitivity, 92.86% specificity, and 97.06% accuracy in HBV infection testing. We anticipate that SATCAS has broad application potential in the early diagnosis, subtyping, drug resistance detection, and point-of-care monitoring of pathogen infectious diseases.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Polimorfismo de Nucleótido Simple / Técnicas de Amplificación de Ácido Nucleico / Sistemas CRISPR-Cas / SARS-CoV-2 Límite: Humans Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Polimorfismo de Nucleótido Simple / Técnicas de Amplificación de Ácido Nucleico / Sistemas CRISPR-Cas / SARS-CoV-2 Límite: Humans Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido