Microchimica Acta

, 186:109 | Cite as

Chronocoulometric aptamer based assay for staphylococcal enterotoxin B by target-triggered assembly of nanostructured dendritic nucleic acids on a gold electrode

  • Xiaoye Chen
  • Yun Liu
  • Yichen Lu
  • Xiong Xiong
  • Yi Li
  • Yuanjian LiuEmail author
  • Xiaohui Xiong
Original Paper


A rapid and ultrasensitive method is described for the detection of staphylococcal enterotoxin B (SEB). It is based on the formation of a dendritic DNA superstructure by integrating (a) target-induced triggering of DNA release with (b) signal amplification by a hybridization chain reaction. Partially complementary pairing of aptamer and trigger DNA forms a duplex structure. The capture DNA is then placed on the surface of a gold electrode through gold-thiol chemistry. In the presence of SEB, the aptamer-target conjugate is compelled to form. This causes the release of trigger DNA owing to a strong competition with SEB. The trigger DNA is subsequently hybridized with the partial complementary sequences of the capture DNA to trigger HCR with three auxiliary DNA sequances (referred to as H1, H2, H3). Finally, the dendritic DNA superstructure is bound to hexaammineruthenium(III) cation by electrostatic adsorption and assembled onto the modified gold electrode. This produces an amplified electrochemical signal that is measured by chronocoulometry. Under optimal conditions, the charge difference increases linearly with the logarithm of the SEB concentrations in the range from 5 pg·mL−1 to 100 ng·mL−1 with a detection limit as low as 3 pg·mL−1 (at S/N = 3).

Graphical abstract

An electrochemical switching strategy is presented for the sensitive detection of Staphylococcus enterotoxin B based on target-triggered assembly of dendritic nucleic acid nanostructures.


Staphylococcus aureus Dendritic nucleic acid nanostructure Hybridization chain reaction Electrochemical nanoprobe ELISA Contaminated food Toxic protein Ruthenium(III) hexammine Electrostatic adsorption 



The project was supported by the National Key Research and Development Program of China (No. 2018YFC1602800), the National Natural Science Foundation of China (No. 21804071), Natural Science Foundation of Jiangsu Province of China (No. BK20180688), and the University Science Research Project of Jiangsu Province in China (16KJB550005).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3236_MOESM1_ESM.docx (112 kb)
ESM 1 (DOCX 112 kb)


  1. 1.
    Singh PK, Agrawal R, Kamboj DV, Gupta G, Boopathi M, Goel AK, Singh L (2010) Construction of a single-chain variable-fragment antibody against the superantigen staphylococcal enterotoxin B. Appl Environ Microbiol 76:8184–8191. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Nunes MM, Caldas ED (2017) Preliminary quantitative microbial risk assessment for staphylococcus enterotoxins in fresh minas cheese, a popular food in Brazil. Food Control 73:524–531. CrossRefGoogle Scholar
  3. 3.
    Fisher EL, Otto M, Cheung GYC (2018) Basis of virulence in enterotoxin-mediated staphylococcal food poisoning. Front Microbiol 9:436. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Wu LY, Gao B, Zhang F, Sun XL, Zhang YZ, Li ZJ (2013) A novel electrochemical immunosensor based on magnetosomes for detection of staphylococcal enterotoxin B in milk. Talanta 106:360–366. CrossRefPubMedGoogle Scholar
  5. 5.
    Pinchuk IV, Beswick EJ, Reyes VE (2010) Staphylococcal enterotoxins. Toxins 2:2177–2197. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Wang WB, Wang WW, Liu LQ, Xu LG, Kuang H, Zhu JP, Xu CL (2016) Nanoshell-enhanced Raman spectroscopy on a microplate for staphylococcal enterotoxin B sensing. ACS Appl Mater Interfaces 8:15591–15597. CrossRefPubMedGoogle Scholar
  7. 7.
    Wu SJ, Duan N, Gu HJ, Hao LL, Ye H, Gong WH, Wang ZP (2016) A review of the methods for detection of staphylococcus aureus enterotoxins. Toxins 8(7).
  8. 8.
    Rodriguez A, Gordillo R, Andrade MJ, Cordoba JJ, Rodriguez M (2016) Development of an efficient real-time PCR assay to quantify enterotoxin-producing staphylococci in meat products. Food Control 60:302–308. CrossRefGoogle Scholar
  9. 9.
    Ludwig S, Jimenez-Bush I, Brigham E, Bose S, Diette G, McCormack MC, Matsui EC, Davis MF (2017) Analysis of home dust for staphylococcus aureus and staphylococcal enterotoxin genes using quantitative PCR. Sci Total Environ 581:750–755. CrossRefPubMedGoogle Scholar
  10. 10.
    Zhou DD, Xie GM, Cao XQ, Chen XP, Zhang X, Chen H (2016) Colorimetric determination of staphylococcal enterotoxin B via DNAzyme-guided growth of gold nanoparticles. Microchim Acta 183:2753–2760. CrossRefGoogle Scholar
  11. 11.
    Mondal B, Ramlal S, Lavu PS, Bhavanashri N, Kingston J (2018) Highly sensitive colorimetric biosensor for staphylococcal enterotoxin B by a label-free aptamer and gold nanoparticles. Front Microbiol 9:179. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Andjelkovic M, Tsilia V, Rajkovic A, De Cremer K, Van Loco J (2016) Application of LC-MS/MS MRM to determine staphylococcal enterotoxins (SEB and SEA) in milk. Toxins 8:118. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Huang YK, Zhang H, Chen XJ, Wang XL, Duan N, Wu SJ, Xu BCWZP (2015) A multicolor time-resolved fluorescence aptasensor for the simultaneous detection of multiplex staphylococcus aureus enterotoxins in the milk. Biosens Bioelectron 74:170–176. CrossRefPubMedGoogle Scholar
  14. 14.
    Wang XL, Huang YK, Wu SJ, Duan N, Xu BC, Wang ZP (2016) Simultaneous detection of staphylococcus aureus and salmonella typhimurium using multicolor time-resolved fluorescence nanoparticles as labels. Int J Food Microbiol 237:172–179. CrossRefPubMedGoogle Scholar
  15. 15.
    Hwang J, Lee S, Choo J (2016) Application of a SERS-based lateral flow immunoassay strip for the rapid and sensitive detection of staphylococcal enterotoxin B. Nanoscale 8:11418–11425. CrossRefPubMedGoogle Scholar
  16. 16.
    Farka Z, Juriik T, Kovaar D, Trnkova L, Sklaadal P (2017) Nanoparticle-based immunochemical biosensors and assays: recent advances and challenges. Chem Rev 117:9973–10042. CrossRefPubMedGoogle Scholar
  17. 17.
    Shahdordizadeh M, Taghdisi SM, Ansari N, Langroodi FA, Abnous K, Ramezani M (2017) Aptamer based biosensors for detection of Staphylococcus aureus. Sensors Actuators B Chem 241:619–635. CrossRefGoogle Scholar
  18. 18.
    Kumari S, Tiwari M, Das P (2019) Multi format compatible visual and fluorometric detection of SEB toxin in nanogram range by carbon dot-DNA and acriflavine nano-assembly. Sensors Actuators B Chem 279:393–399. CrossRefGoogle Scholar
  19. 19.
    Chen XY, Shi XP, Liu Y, Lu LX, Lu YC, Xiong X, Liu YJ, Xiong XH (2018) Impedimetric determination of staphylococcal enterotoxin B using electrochemical switching with DNA triangular pyramid frustum nanostructure. Microchim Acta 185:460. CrossRefGoogle Scholar
  20. 20.
    Hou T, Li W, Liu XJ, Li F (2015) Label-free and enzyme-free homogeneous electrochemical biosensing strategy based on hybridization chain reaction: a facile, sensitive, and highly specific microRNA assay. Anal Chem 87:11368–11374. CrossRefPubMedGoogle Scholar
  21. 21.
    Ge L, Wang WX, Sun XM, Hou T, Li F (2016) Affinity-mediated homogeneous electrochemical aptasensor on a graphene platform for ultrasensitive biomolecule detection via exonuclease-assisted target-analog recycling amplification. Anal Chem 88:2212–2219. CrossRefPubMedGoogle Scholar
  22. 22.
    Wang XZ, Jiang AW, Hou T, Li HY, Li F (2015) Enzyme-free and label-free fluorescence aptasensing strategy for highly sensitive detection of protein based on target-triggered hybridization chain reaction amplification. Biosens Bioelectron 70:324–329. CrossRefPubMedGoogle Scholar
  23. 23.
    Huang DJ, Huang ZM, Xiao HY, Wu ZK, Tang LJ, Jiang JH (2018) Protein scaffolded DNA tetrads enable efficient delivery and ultrasensitive imaging of miRNA through crosslinking hybridization chain reaction. Chem Sci 9:4892–4897. CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Yan MM, Bai WH, Zhu C, Huang YF, Yan J, Chen AL (2017) Design of nuclease-based target recycling signal amplification in aptasensors. Biosens Bioelectron 77:613–623. CrossRefGoogle Scholar
  25. 25.
    Shuai HL, Wu X, Huang KJ, Zhai ZB (2017) Ultrasensitive electrochemical biosensing platform based on spherical silicon dioxide/molybdenum selenide nanohybrids and triggered hybridization chain reaction. Biosens Bioelectron 94:616–625. CrossRefPubMedGoogle Scholar
  26. 26.
    Yang DW, Tang YG, Miao P (2017) Hybridization chain reaction directed DNA superstructures assembly for biosensing applications. TrAC Trends Anal Chem 94:1–13. CrossRefGoogle Scholar
  27. 27.
    Shimron S, Wang F, Orbach R, Willner I (2012) Amplified detection of DNA through the enzyme-free autonomous assembly of hemin/G-quadruplex DNAzyme nanowires. Anal Chem 84:1042–1048. CrossRefPubMedGoogle Scholar
  28. 28.
    Li CX, Wang HM, Shang JH, Liu XQ, Yuan BF, Wang F (2018) Highly sensitive assay of methyltransferase activity based on an autonomous concatenated DNA circuit. ACS Sens 3:2359–2366. CrossRefPubMedGoogle Scholar
  29. 29.
    Xu N, Lei JP, Wang QB, Yang QH, Ju HX (2016) Dendritic DNA-porphyrin as mimetic enzyme for amplified fluorescent detection of DNA. Talanta 150:661–665. CrossRefPubMedGoogle Scholar
  30. 30.
    Temur E, Zengin A, Boyaci IH, Dudak FC, Torul H, Tamer U (2012) Attomole sensitivity of staphylococcal enterotoxin B detection using an aptamer-modified surface-enhanced Raman scattering probe. Anal Chem 84:10600–10606. CrossRefPubMedGoogle Scholar
  31. 31.
    Han JH, Kim HJ, Sudheendra L, Gee SJ, Hammock BD, Kennedy IM (2013) Photonic crystal lab-on-a-chip for detecting staphylococcal enterotoxin B at low attomolar concentration. Anal Chem 85:3104–3109. CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Xiong XH, Shi XP, Liu YJ, Lu LX, You JJ (2018) An aptamer-based electrochemical biosensor for simple and sensitive detection of staphylococcal enterotoxin B in milk. Anal Methods 10:365–370. CrossRefGoogle Scholar
  33. 33.
    Mudili V, Makam SS, Sundararaj N, Siddaiah C, Gupta VK, Rao PVL (2015) A novel IgY-aptamer hybrid system for cost-effective detection of SEB and its evaluation on food and clinical samples. Sci Rep 5:15151. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Coll Food Sci & Light IndNanjing Tech UniversityNanjingPeople’s Republic of China

Personalised recommendations