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author:

Xu, Huo (Xu, Huo.) [1] | Wu, Dong (Wu, Dong.) [2] | Li, Chen-Qiao (Li, Chen-Qiao.) [3] | Lu, Zheng (Lu, Zheng.) [4] | Liao, Xiao-Yun (Liao, Xiao-Yun.) [5] | Huang, Jie (Huang, Jie.) [6] | Wu, Zai-Sheng (Wu, Zai-Sheng.) [7] (Scholars:吴再生)

Indexed by:

EI Scopus SCIE

Abstract:

Highly sensitive detection of K-ras gene is of great significance in biomedical research and clinical diagnosis. Here, we developed a colorimetric biosensing system for the detection of proto-oncogene K-ras based on enhanced amplification effect of DNA molecular machine, where dual isothermal circular strand-displacement amplification (D-SDA) occurs on two arms in one-to-one correspondence. Specifically, we designed a primer locked hairpin probe (HP) and a primer-contained linear polymerization template (PPT). In the presence of target gene, HP can hybridize with PPT, forming a DNA molecular machine with dual functional arms (called DFA-machine). Each of the two probes in this machine is able to be extended by polymerase on its counterpart species. Moreover, with the help of nicking endonuclease, the dual isothermal polymerization is converted into dual circular strand-displacement amplification, generating a large amount of anti-hemin aptamer-contained products. After binding to hemins, the aptamer/hemin duplex, horseradish peroxidase (HRP)-mimicking DNAzyme, was formed and catalyzed the oxidation of colorless ABTS by H2O2, producing a visible green color. The proposed colorimetric assay exhibits a wide linear range from 0.01 to 150 nM with a low detection limit of 10 pM. More interestingly, the mutations existing in target gene are easily observed by the naked eye. It should be noted that this calorimetric system was proved by the analysis of K-ras gene of SW620 cell lines. The simple and powerful DFA-machine is expected to provide promising potential in the sensitive detection of biomarkers for cancer diagnosis, prognosis and therapy.

Keyword:

Colorimetric detection DNA molecular machine with dual functional arms (DFA-machine) DNAzyme Dual isothermal circular strand-displacement amplification (D-SDA) K-ras gene

Community:

  • [ 1 ] [Wu, Zai-Sheng]Fuzhou Univ, Canc Metastasis Alert & Prevent Ctr, Pharmaceut Photocatalysis State Key Lab Photocata, Coll Chem, Fuzhou 350002, Peoples R China
  • [ 2 ] [Wu, Zai-Sheng]Fuzhou Univ, Fujian Prov Key Lab Canc Metastasis Chemoprevent, Coll Chem, Fuzhou 350002, Peoples R China

Reprint 's Address:

  • 吴再生

    [Wu, Zai-Sheng]Fuzhou Univ, Canc Metastasis Alert & Prevent Ctr, Pharmaceut Photocatalysis State Key Lab Photocata, Coll Chem, Fuzhou 350002, Peoples R China

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Source :

BIOSENSORS & BIOELECTRONICS

ISSN: 0956-5663

Year: 2017

Volume: 90

Page: 314-320

8 . 1 7 3

JCR@2017

1 0 . 7 0 0

JCR@2023

ESI Discipline: CHEMISTRY;

ESI HC Threshold:226

JCR Journal Grade:1

CAS Journal Grade:1

Cited Count:

WoS CC Cited Count: 44

SCOPUS Cited Count: 49

ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 1

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