Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 10, pp 2101–2109 | Cite as

Facilely prepared low-density DNA monolayer–based electrochemical biosensor with high detection performance in human serum

  • Jinyuan Chen
  • Chenliu Ye
  • Zhoujie Liu
  • Liangyong Yang
  • Ailin Liu
  • Guangxian ZhongEmail author
  • Huaping PengEmail author
  • Xinhua LinEmail author
Research Paper
  • 113 Downloads

Abstract

Presently, most reported electrochemical biosensors, for highly sensitive and selective detection of nucleic acid, still require multiple, time-consuming assembly steps and high-consumption DNA probes as well as lack good performance in human serum, which greatly limit their applicability. Herein, an easy-to-fabricate electrochemical DNA biosensor constructed by assembly of bovine serum albumin (BSA) followed with direct incubation of amplified products has been proposed. This method combined terminal deoxynucleoside transferase (TdTase)–mediated isothermal amplification and polyHRP catalysis to achieve dual-signal enhancement, and was featured with low-density DNA monolayer for its employment of only 2 nM capture probes. Surprisingly, based on the low-density DNA monolayer, the steric hindrance effect of polyHRP could effectively restrain the background compared with HRP, which further pushes the signal-to-noise (S/N) ratio to 70 than that of most currently available methods. Additionally, this strategy also showed favorable specificity and powerful anti-interference in human serum, and thus potentially attractive for diagnosis of diseases.

Keywords

Electrochemical DNA biosensor Isothermal amplification Steric hindrance effect Low-density DNA monolayer 

Notes

Funding information

This work was financially supported by the National Natural Science Foundation of China (21775023); the Medical Elite Cultivation Program of Fujian Province (2018-ZQN-49); the Open Program for the Key Lab/Research Platform of the First Affiliated Hospital of Fujian Medical University (FYKFKT-201707); the Outstanding Youth Scientific Research Personnel Training Plan of Colleges and Universities in Fujian Province (2015B027); Joint Funds for the Innovation of Science and Technology in Fujian Province (2016Y9019); and the Special Fund of Youth Top-Notch Innovative Talents of Fujian Province (SQNBJ201601).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical standards and informed consent

This study conformed to the ethical guidelines of the Declaration of Helsinki and was approved by The Ethics Committee for Human Research, The First Affiliated Hospital of Fujian Medical University. Human serum samples used in this study do not have any identifying information about all the participants that provided written informed consent.

Supplementary material

216_2019_1637_MOESM1_ESM.pdf (389 kb)
ESM 1 (PDF 388 kb)

References

  1. 1.
    Zhang HL, Liu XH, Liu MH, Gao T, Huang YZ, Liu Y, et al. Gene detection: an essential process to precision medicine. Biosens Bioelectron. 2018;99:625–36.CrossRefGoogle Scholar
  2. 2.
    Choi JR, Hu J, Tang RH, Gong Y, Feng SS, Ren H, et al. An integrated paper-based sample-to-answer biosensor for nucleic acid testing at the point of care. Lab Chip. 2016;16:611–21.CrossRefGoogle Scholar
  3. 3.
    Wang YQ, Qu JX, Ba Q, Dong JH, Zhang L, Zhang H, et al. Detection and typing of human-infecting influenza viruses in China by using a multiplex DNA biochip assay. J Virol Methods. 2016;234:178–85.CrossRefGoogle Scholar
  4. 4.
    Dong J, Chen GY, Wang W, Huang X, Peng HP, Pu QL, et al. Colorimetric PCR-based microRNA detection method based on small organic dye and single enzyme. Anal Chem. 2018;90:7107–11.CrossRefGoogle Scholar
  5. 5.
    Jae-Ho K, Jeong-Eun P, Mouhong L, Sungi K, Gyeong-Hwan K, SungJun P, et al. Sensitive, quantitative naked-eye biodetection with polyhedral Cu nanoshells. Adv Mater. 2017;29:1702945.CrossRefGoogle Scholar
  6. 6.
    Ma Q, Gao ZQ. A simple and ultrasensitive fluorescence assay for single-nucleotide polymorphism. Anal Bioanal Chem. 2018;410:3093–100.CrossRefGoogle Scholar
  7. 7.
    Nano A, Boynton AN, Barton JK. A rhodium-cyanine fluorescent probe: detection and signaling of mismatches in DNA. J Am Chem Soc. 2017;139:17301–4.CrossRefGoogle Scholar
  8. 8.
    Park S, Jeong J-E, Le VS, Seo J, Yu B, Kim D-Y, et al. Enhanced electron transfer mediated by conjugated polyelectrolyte and its application to washing-free DNA detection. J Am Chem Soc. 2018;140:2409–12.CrossRefGoogle Scholar
  9. 9.
    Wang S, Zhang LQ, Wan S, Cansiz S, Cui C, Liu Y, et al. Aptasensor with expanded nucleotide using DNA nanotetrahedra for electrochemical detection of cancerous exosomes. ACS Nano. 2017;11:3943–9.CrossRefGoogle Scholar
  10. 10.
    Hou T, Liu XJ, Wang XZ, Jiang AW, Liu SF, Li F. DNAzyme-guided polymerization of aniline for ultrasensitive electrochemical detection of nucleic acid with bio-bar codes-initiated rolling circle amplification. Sensors Actuators B Chem. 2014;190:384–8.CrossRefGoogle Scholar
  11. 11.
    Chen AY, Ma SY, Zhuo Y, Chai YQ, Yuan R. In situ electrochemical generation of electrochemiluminescent silver naonoclusters on target-cycling synchronized rolling circle amplification platform for microRNA detection. Anal Chem. 2016;88:3203–10.CrossRefGoogle Scholar
  12. 12.
    Xie SB, Yuan YL, Chai YQ, Yuan R. Tracing phosphate ions generated during loop-mediated isothermal amplification for electrochemical detection of nosema bombycis genomic DNA PTP1. Anal Chem. 2015;87:10268–74.CrossRefGoogle Scholar
  13. 13.
    Hsieh K, Patterson AS, Scott FB, Plaxco KW, Soh HT. Rapid, sensitive, and quantitative detection of pathogenic DNA at the point of care through microfluidic electrochemical quantitative loop-mediated isothermal amplification. Angew Chem Int Ed. 2012;51:4896–900.CrossRefGoogle Scholar
  14. 14.
    Del Río JS, Svobodova M, Bustos P, Conejeros P, O'Sullivan CK. Electrochemical detection of Piscirickettsia salmonis genomic DNA from salmon samples using solid-phase recombinase polymerase amplification. Anal Bioanal Chem. 2016;408:8611–20.CrossRefGoogle Scholar
  15. 15.
    Fang CS, Kim K-S, Ha DT, Kim M-S, Yang H. Washing-free electrochemical detection of amplified double-stranded DNAs using a zinc finger protein. Anal Chem. 2018;90:4776–82.CrossRefGoogle Scholar
  16. 16.
    Yu YY, Chen ZG, Jian WS, Sun DP, Zhang BB, Li XC, et al. Ultrasensitive electrochemical detection of avian influenza A (H7N9) virus DNA based on isothermal exponential amplification coupled with hybridization chain reaction of DNAzyme nanowires. Biosens Bioelectron. 2015;64:566–71.CrossRefGoogle Scholar
  17. 17.
    Wang D, Chai YQ, Yuan YL, Yuan R. A peptide cleavage-based ultrasensitive electrochemical biosensor with an ingenious two-stage DNA template for highly efficient DNA exponential amplification. Anal Chem. 2017;89:8951–6.CrossRefGoogle Scholar
  18. 18.
    Chow DC, Lee W-K, Zauscher S, Chilkoti A. Enzymatic fabrication of DNA nanostructures: extension of a self-assembled oligonucleotide monolayer on gold arrays. J Am Chem Soc. 2005;127:14122–3.CrossRefGoogle Scholar
  19. 19.
    Yang F, Yang X, Wang YZ, Qin Y, Liu X, Yan XQ, et al. Template-independent, in situ grown DNA nanotail enabling label-free femtomolar chronocoulometric detection of nucleic acids. Anal Chem. 2014;86:11905–12.CrossRefGoogle Scholar
  20. 20.
    Chen JY, Liu ZJ, Peng HP, Zheng YJ, Lin Z, Liu AL, et al. Electrochemical DNA biosensor based on grafting-to mode of terminal deoxynucleoside transferase-mediated extension. Biosens Bioelectron. 2017;98:345–9.CrossRefGoogle Scholar
  21. 21.
    Liu YH, Li HN, Chen W, Liu AL, Lin XH, Chen YZ. Bovine serum albumin-based probe carrier platform for electrochemical DNA biosensing. Anal Chem. 2013;85:273–7.CrossRefGoogle Scholar
  22. 22.
    Wan Y, Xu H, Su Y, Zhu XH, Song SP, Fan CH. A surface-initiated enzymatic polymerization strategy for electrochemical DNA sensors. Biosens Bioelectron. 2013;41:526–31.CrossRefGoogle Scholar
  23. 23.
    Ge ZL, Lin MH, Wang P, Pei H, Yan J, Shi JY, et al. Hybridization chain reaction amplification of microRNA detection with a tetrahedral DNA nanostructure-based electrochemical biosensor. Anal Chem. 2014;86:2124–30.CrossRefGoogle Scholar
  24. 24.
    Wan Y, Wang PJ, Su Y, Wang LH, Pan D, Aldalbahi A, et al. Nanoprobe-initiated enzymatic polymerization for highly sensitive electrochemical DNA detection. ACS Appl Mat Interfaces. 2015;7:25618–23.CrossRefGoogle Scholar
  25. 25.
    Li C, Wu D, Hu XL, Xiang Y, Shu YQ, Li GX. One-step modification of electrode surface for ultrasensitive and highly selective detection of nucleic acids with practical applications. Anal Chem. 2016;88:7583–90.CrossRefGoogle Scholar
  26. 26.
    Song P, Li M, Shen JW, Pei H, Chao J, Su S, et al. Dynamic modulation of DNA hybridization using allosteric DNA tetrahedral nanostructures. Anal Chem. 2016;88:8043–9.CrossRefGoogle Scholar
  27. 27.
    Li LY, Wang LL, Xu Q, Xu L, Liang W, Li Y, et al. Bacterial analysis using an electrochemical DNA biosensor with poly-adenine-mediated DNA self-assembly. ACS Appl Mat Interfaces. 2018;10:6895–903.CrossRefGoogle Scholar
  28. 28.
    Wu J, Campuzano S, Halford C, Haake DA, Wang J. Ternary surface monolayers for ultrasensitive (zeptomole) amperometric detection of nucleic acid hybridization without signal amplification. Anal Chem. 2010;82:8830–7.CrossRefGoogle Scholar
  29. 29.
    Campuzano S, Kuralay F, Lobo-Castañón MJ, Bartošík M, Vyavahare K, Paleček E, et al. Ternary monolayers as DNA recognition interfaces for direct and sensitive electrochemical detection in untreated clinical samples. Biosens Bioelectron. 2011;26:3577–83.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The CentralabThe First Affiliated Hospital of Fujian Medical UniversityFuzhouChina
  2. 2.Department of Pharmaceutical Analysis, Faculty of PharmacyFujian Medical UniversityFuzhouChina
  3. 3.Department of OrthopaedicsThe First Affiliated Hospital of Fujian Medical UniversityFuzhouChina

Personalised recommendations