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author:

Zhu, Shengting (Zhu, Shengting.) [1] | Chen, Jianlin (Chen, Jianlin.) [2] | Yu, Lili (Yu, Lili.) [3] | Li, Jiawen (Li, Jiawen.) [4] | You, Shumin (You, Shumin.) [5] | Zheng, Yue (Zheng, Yue.) [6] | Zhuang, Wanzhen (Zhuang, Wanzhen.) [7] | Qiu, Bin (Qiu, Bin.) [8] (Scholars:邱彬) | Huang, Yi (Huang, Yi.) [9]

Indexed by:

Scopus SCIE

Abstract:

Our preliminary investigation has identified the potential of serum fucosylated extracellular vesicles (EVs) miR-4732-5p in the early diagnosis of lung adenocarcinoma (LUAD) by a fucose-captured strategy utilizing lentil lectin (LCA)-magnetic beads and subsequent screening of high throughput sequencing and validation of real-time quantitative polymerase chain reaction (RT-qPCR). Considering the relatively complicated procedure, expensive equipment, and stringent laboratory condition, we have constructed an electrochemical biosensor assay for the detection of miR-4732-5p. miR-4732-5p is extremely low in serum, down to the fM level, so it needs to be detected by highly sensitive electrochemical methods based on the Mg2+-dependent DNAzyme splitting nucleic acid lock (NAL) cycle and hybridization chain reaction (HCR) signal amplification. In this study, signal amplification is achieved through the dual amplification reactions using NAL cycle in combination with HCR. In addition, hybridized DNA strands bind to a large number of methylene blue (MB) molecules to enhance signaling. Based on the above strategy, we further enhance our signal amplification strategies to improve detection sensitivity and accuracy. The implementation of this assay proceeded as follows: initially, miR-4732-5p was combined with NAL, and then Mg2+-dependent DNAzyme splitted NAL to release auxiliary DNA (S1) strands, which were subsequently captured by the immobilized capture probe DNA (C1) strands on the electrode surface. Following this, abundant quantities of DNA1 (H1) and DNA2 (H2) tandems were generated by HCR, and S1 strands then hybridized with the H1 and H2 tandems through base complementary pairing. Finally, MB was bonded to the H1 and H2 tandems through pi-pi stacking interaction, leading to the generation of a signal current upon the detection of a potential capable of inducing a redox change of MB by the electrode. Furthermore, we evaluated the performance of our developed electrochemical biosensor assay. The results demonstrated that our assay is a reliable approach, characterized by its high sensitivity (with a detection limit of 2.6x10(-17) M), excellent specificity, good accuracy, reproducibility, and stability. Additionally, it is cost-effective, requires simple operation, and is portable, making it suitable for the detection of serum fucosylated extracellular vesicles miR-4732-5p. Ultimately, this development has the potential to enhance the diagnostic efficiency for patients with early-stage LUAD.

Keyword:

Biomarker Diagnostic Electrochemical biosensor Lung adenocarcinoma miR-4732-5p

Community:

  • [ 1 ] [Zhu, Shengting]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 2 ] [Chen, Jianlin]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 3 ] [Yu, Lili]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 4 ] [Zheng, Yue]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 5 ] [Zhuang, Wanzhen]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 6 ] [Huang, Yi]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China
  • [ 7 ] [Zhu, Shengting]Fujian Prov Hosp, Dept Blood Transfus, Fuzhou 350001, Peoples R China
  • [ 8 ] [Li, Jiawen]Fuzhou Univ, Fujian Prov Key Lab Anal & Detect Food Safety, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou 350108, Peoples R China
  • [ 9 ] [You, Shumin]Fuzhou Univ, Fujian Prov Key Lab Anal & Detect Food Safety, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou 350108, Peoples R China
  • [ 10 ] [Qiu, Bin]Fuzhou Univ, Fujian Prov Key Lab Anal & Detect Food Safety, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou 350108, Peoples R China
  • [ 11 ] [Chen, Jianlin]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China
  • [ 12 ] [Yu, Lili]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China
  • [ 13 ] [Zheng, Yue]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China
  • [ 14 ] [Zhuang, Wanzhen]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China
  • [ 15 ] [Huang, Yi]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China
  • [ 16 ] [Huang, Yi]Fujian Prov Hosp, Cent Lab, Ctr Expt Res Clin Med, Fuzhou 350001, Peoples R China
  • [ 17 ] [Huang, Yi]Fujian Prov Key Lab Cardiovasc Dis, Fujian Prov Key Lab Crit Care Med, Fuzhou 350001, Peoples R China

Reprint 's Address:

  • [Huang, Yi]Fujian Med Univ, Shengli Clin Med Coll, Fuzhou 350001, Peoples R China;;[Qiu, Bin]Fuzhou Univ, Fujian Prov Key Lab Anal & Detect Food Safety, Key Lab Analyt Sci Food Safety & Biol, Minist Educ, Fuzhou 350108, Peoples R China;;[Huang, Yi]Fujian Prov Hosp, Dept Clin Lab, Fuzhou 350001, Peoples R China;;[Huang, Yi]Fujian Prov Hosp, Cent Lab, Ctr Expt Res Clin Med, Fuzhou 350001, Peoples R China;;[Huang, Yi]Fujian Prov Key Lab Cardiovasc Dis, Fujian Prov Key Lab Crit Care Med, Fuzhou 350001, Peoples R China;;

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Source :

SCIENTIFIC REPORTS

ISSN: 2045-2322

Year: 2024

Issue: 1

Volume: 14

3 . 8 0 0

JCR@2023

Cited Count:

WoS CC Cited Count:

SCOPUS Cited Count:

ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 3

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