Citation: WANG Hai-Fei, TANG Ying, YAN Xiao-Mei, CHEN Zeng-Ping, CHEN Yao. Exonuclease Ⅲ and Gold Substrate Capturing Cyclic Strategy for Detection of Manganese Superoxide Dismutase DNA[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(5): 720-727. doi: 10.19756/j.issn.0253-3820.210740 shu

Exonuclease Ⅲ and Gold Substrate Capturing Cyclic Strategy for Detection of Manganese Superoxide Dismutase DNA

WANG Hai-Fei ¹ ,
TANG Ying ^1,2,, ,
YAN Xiao-Mei ² ,
CHEN Zeng-Ping ² ,
CHEN Yao ^1,2,,

1.
Hunan Key Lab of Biomedical Materials and Devices, College of Life Sciences and Chemistry, Hunan University of Technology, Zhuzhou 412007, China;
2.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China

Corresponding author: TANG Ying, CHEN Yao,
Received Date: 13 September 2021
Revised Date: 1 December 2021

Fund Project: Supported by the National Natural Science Foundation of China (Nos.21705044, 21804040), the Natural Science Foundation of Hunan Province, China (Nos.2018JJ3114, 2019JJ50134), the Scientific Research Project of the Education Department of Hunan Province, China (No.19C0554) and the Open Project of the State Key Laboratory of Chemical and Biological Sensing and Metrology (No.2018015).

A sensitive and simple method was developed for detection of manganese superoxide dismutase (MnSOD) DNA using exonuclease Ⅲ assisted signal amplification technique and gold substrate capture cycle strategy. This strategy relied on three major steps in the experiment. First, the hairpin probe labeled with fluorophore was opened by MnSOD DNA. Second, with the help of exonuclease Ⅲ, the hairpin probe was partially hydrolyzed to release MnSOD DNA and residual hairpin probe fragments labeled with fluorophore. The released MnSOD DNA could be recycled and continued to hybridize with the complete hairpin probe. And third, the reaction solution contained a large number of residual hairpin probe fragments labeled with fluorophore, which could be hybridized with capture probe modified on gold substrate, and then transferred to a new tube solution for fluorescence detection. Based on this principle, the method for MnSOD DNA detection was constructed, with detection limit of about 0.42 pmol/L. The method was also applied to the detection of MnSOD DNA in lysates of HeLa, Jurkat and Ramos cells, with recoveries ranging from 90.2% to 111.2%. This method could effectively eliminate the influence of background interference and matrix effect, and provide a new promising paradigm in developing universal methods for detection of short DNA.
- Exonuclease Ⅲ,
- Gold substrate,
- Manganese superoxide dismutase DNA,
- Quantitative detection
1. [1]
  MA S, FU A, LIM S, GIAP G, CHIEW Y, LUO K Q. Free Radical Bio. Med., 2018, 129:46-58.
2. [2]
  OBERLEY L W. Biomed. Pharmacother., 2005, 59(4):143-148.
3. [3]
  MAO Y, QIAO J D, CHEN S, ZHOU X, WANG Z, CAI S, LI L, LUO Y. Reprod. Sci., 2021, 28(4):1012-1019.
4. [4]
  MARTINS D, MCKAY G, SAMPATHKUMAR G, KHAKIMOVA M, ENGLISH A M, NGUYEN D. Proc. Natl. Acad. Sci. U. S. A., 2018, 115(39):9797-9802.
5. [5]
  WARD M B, SCHEITLER A, YU M, SENFT L, ZILLMANN A S, GORDEN J D, SCHWARTZ D D, IVANOVIC-BURMAZOVIC I, GOLDSMITH C R. Nat. Chem., 2018, 10(12):1207-1212.
6. [6]
  MACMILLAN-CROW L A, CRUTHIRDS D L. Free Radical Res., 2001, 34(4):325-336.
7. [7]
  TEOH-FITZGERALD M L T, FITZGERALD M P, JENSEN T J, FUTSCHER B W, DOMANN F E. Mol. Cancer Res., 2012, 10(1):40-51.
8. [8]
  GRIESS B, TOM E, DOMANN F, TEOH-FITZGERALD M. Free Radical Bio. Med., 2017, 112:464-479.
9. [9]
  LIU Y D, YU L, YING L, BALIC J, GAO H, DENG N T, WEST A, YAN F, JI C B, GOUGH D, TAN P, JENKINS B J, LI J K. Int. J. Cancer, 2019, 144(12):3056-3069.
10. [10]
  LI Y, ZHAO Q, WANG Y, MAN T, ZHOU L, FANG X, PEI H, CHI L, LIU J. Anal. Chem., 2016, 88(23):11684-11690.
11. [11]
  ZHENG X, LI L, ZHANG L, XIE L, SONG S, YU J. Biosens. Bioelectron., 2020, 147(1):111769.
12. [12]
  LU C H, LI J, LIN M H, WANG Y W, YANG H H, CHEN X, CHEN G N. Angew. Chem., Int. Ed., 2010, 122(45):8632-8635.
13. [13]
  ZUO X, XIA F, XIAO Y, PLAXCO K W. J. Am. Chem. Soc., 2010, 132(6):1816-1818.
14. [14]
  ZHANG K, YANG L, LU F, WU X, ZHU J J. Small, 2018, 14(10):1703858.
15. [15]
  WANG L, DONG L, LIU G, SHEN X, WANG J, ZHU C, DING M, WEN Y. Microchim. Acta, 2019, 186(5):1-6.
16. [16]
  LI Y, ZHAO Q, WANG Y, MAN T, ZHOU L, FANG X, PEI H, CHI L, LIU J. Anal. Chem., 2016, 88(23):11684-11690.
17. [17]
  BAO M, JENSEN E, CHANG Y, KORENKY G, DU K. ACS Appl. Mater. Interfaces, 2020, 12(39):43435-43443.
18. [18]
19. [19]
  ALI Z, WANG J, TANG Y, LIU B, HE N, LI Z. Biomater. Sci., 2017, 5(1):57-66.
20. [20]
  GARG N, CARRASQUILLO-MOLINA E, LEE T R. Langmuir, 2002, 18(7):2717-2726.
21. [21]
  CUI L, KE G, ZHANG W Y, YANG C J. Biosens. Bioelectron., 2011, 26(5):2796-2800.
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