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A CRISPR/Cas13a-powered catalytic electrochemical biosensor for successive and highly sensitive RNA diagnostics.
Sheng, Yan; Zhang, Tenghua; Zhang, Shihong; Johnston, Midori; Zheng, Xiaohe; Shan, Yuanyue; Liu, Tong; Huang, Zena; Qian, Feiyang; Xie, Zihui; Ai, Yiru; Zhong, Hankang; Kuang, Tairong; Dincer, Can; Urban, Gerald Anton; Hu, Jiaming.
Afiliación
  • Sheng Y; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Zhang T; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Zhang S; Department of Clinical Laboratory, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China.
  • Johnston M; Freiburg Center for Interactive Materials and Bioinspired Technologies - FIT, University of Freiburg, Georges-Koehler Allee 105, 79110, Freiburg, Germany; Department of Microsystems Engineering - IMTEK, Laboratory for Sensors, University of Freiburg, Georges-Koehler Allee 103, 79110, Freiburg, Germa
  • Zheng X; Department of Clinical Laboratory, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China.
  • Shan Y; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Liu T; College of Materials Science and Engineering, Zhejiang University of Technology, Hangzhou, 310014, China.
  • Huang Z; Department of General Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, China.
  • Qian F; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Xie Z; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Ai Y; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Zhong H; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China.
  • Kuang T; College of Materials Science and Engineering, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: kuangtr@zjut.edu.cn.
  • Dincer C; Freiburg Center for Interactive Materials and Bioinspired Technologies - FIT, University of Freiburg, Georges-Koehler Allee 105, 79110, Freiburg, Germany; Department of Microsystems Engineering - IMTEK, Laboratory for Sensors, University of Freiburg, Georges-Koehler Allee 103, 79110, Freiburg, Germa
  • Urban GA; Department of Microsystems Engineering - IMTEK, Laboratory for Sensors, University of Freiburg, Georges-Koehler Allee 103, 79110, Freiburg, Germany.
  • Hu J; MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, South China Normal University, Guangzhou, 510631, China; College of Biophotonics, South China Normal University, Guangzhou, 510631, China. Electronic address: jm
Biosens Bioelectron ; 178: 113027, 2021 Apr 15.
Article en En | MEDLINE | ID: mdl-33529861
Rapid and specific quantitation of a variety of RNAs with low expression levels in early-stage cancer is highly desirable but remains a challenge. Here, we present a dual signal amplification strategy consisting of the CRISPR/Cas13a system and a catalytic hairpin DNA circuit (CHDC), integrated on a reusable electrochemical biosensor for rapid and accurate detection of RNAs. Signal amplification is accomplished through the unique combination of the CRISPR/Cas13a system with CHDC, achieving a limit of detection of 50 aM within a readout time of 6 min and an overall process time of 36 min, using a measuring volume of 10 µL. Enzymatic regeneration of the sensor surface and ratiometric correction of background signal allow up to 37 sequential RNA quantifications by square-wave voltammetry on a single biosensor chip without loss of sensitivity. The reusable biosensor platform could selectively (specificity = 0.952) and sensitively (sensitivity = 0.900) identify low expression RNA targets in human serum, distinguishing early-stage patients (n = 20) suffering from non-small-cell lung carcinoma (NSCLC) from healthy subjects (n = 30) and patients with benign lung disease (n = 12). Measurement of six NSCLC-related RNAs (miR-17, miR-155, TTF-1 mRNA, miR-19b, miR-210 and EGFR mRNA) shows the ability of the electrochemical CRISPR/CHDC system to be a fast, low-cost and highly accurate tool for early cancer diagnostics.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Carcinoma de Pulmón de Células no Pequeñas / ADN Catalítico / Neoplasias Pulmonares Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2021 Tipo del documento: Article País de afiliación: China

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Carcinoma de Pulmón de Células no Pequeñas / ADN Catalítico / Neoplasias Pulmonares Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Humans Idioma: En Revista: Biosens Bioelectron Asunto de la revista: BIOTECNOLOGIA Año: 2021 Tipo del documento: Article País de afiliación: China
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