A light-up fluorescence platform based DNA: RNA hybrid G-quadruplet for detecting single nucleotide variant of ctDNA and miRNA-21

  • Talanta. 2023 May 15:257:124373. doi: 10.1016/j.talanta.2023.124373.
Gaihua Cao  1 ,  Keyi Long  1 ,  Yue Qiu  1 ,  Lan Peng  2 ,  Haiyan Han  3 ,  Yongzhong Wang  4 ,  Danqun Huo  5 ,  Changjun Hou  6
Affiliations
  • 1. Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China.
  • 2. Chongqing Medical and Pharmaceutical College Basic Department, Chongqing, 401331, PR China.
  • 3. Chongqing Institute for Food and Drug Control, Chongqing, 401121, PR China.
  • 4. Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China. Electronic address: [email protected].
  • 5. Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China; Chongqing Key Laboratory of Bio-perception & Intelligent Information Processing, School of Microelectronics and Communication Engineering, Chongqing University, Chongqing, 400044, PR China. Electronic address: [email protected].
  • 6. Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China. Electronic address: [email protected].
PMID: 36801760 DOI: 10.1016/j.talanta.2023.124373
Abstract

The nucleic acid assay is an area of great concern in the diagnosis and treatment of breast Cancer. Here, we developed a DNA: RNA hybrid G-quadruplet (HQ) detection platform based on strand displacement amplification (SDA) and Baby Spinach RNA aptamer for single nucleotide variant (SNV) of circulating tumor DNA (ctDNA) and miRNA-21. This was the first in vitro construction of HQ for the biosensor. It found that HQ had much stronger ability to switch on fluorescence of DFHBI-1T than Baby Spinach RNA alone. Taking advantage of the platform and the FspI enzyme with high specificity, the biosensor achieved ultra-sensitive detection of SNV of the ctDNA (PIK3CA H1047R gene) and miRNA-21. The light-up biosensor had high anti-interference ability in complex actual samples. Hence, the label-free biosensor provided a sensitive and accurate method for early diagnosis of breast Cancer. Moreover, it opened a new application model for RNA Aptamers.

Keywords
Circulating tumor DNA (ctDNA); Fluorescence; G-quadruplet; Strand displacement amplification (SDA); miRNA.
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