Microchimica Acta

, 186:155 | Cite as

Sensitive fluorometric determination of platelet-derived growth factor BB and avian influenza A virus DNA via dual signal amplification using the hybridization chain reaction and glucose oxidase assisted recycling

  • Yubin LiEmail author
  • Jing Shao
  • Wanting Guo
  • Minting Wang
Original Paper


A method is described for fluorometric determination of platelet-derived growth factor BB (PDGF-BB) and avian influenza A (H1N1) virus DNA. It is based on the use of the hybridization chain reaction (HCR) and of glucose oxidase (GOx) assisted dual-recycling amplification. A silver coated glass slide (SCGS) serves as an ideal material for separation. A signal DNA/initiator triggers the HCR and generates a cascade of hybridization to form a nicked double-helix polymer. Upon addition of the analytes (PDGF-BB or H1N1 DNA) and capture DNA immobilized on the SCGS, the nicked double-helix polymer binds on the surface of the SCGS through formation of a [capture DNA/analyte/signal DNA] sandwich structure. The GOx-biotin-streptavidin (SA) complexes were then attached to the nicked double-helix polymer through SA-biotin interaction. After cleavage by DNase I, the bound GOx is transferred into the buffer. Glucose is added and enzymatically oxidized to produce H2O2. The H2O2 formed oxidizes the substrate 3-(p-hydroxyphenyl)-propanoic acid to give a blue fluorescent product (with excitation/emission maxima at 320/416 nm) under the catalysis of horseradish peroxidase. Under optimal conditions, fluorescence increases linearly in the 0.5 to 70 pmol·L−1 PDGF-BB concentration range, and the detection limit is 191 fmol·L−1. For the H1N1 virus DNA, the respective data are 2.5 to 300 pmol·L−1 and 826 fmol·L−1.

Graphical abstract

Schematic presentation for detection of analytes (PDGF-BB or H1N1 virus DNA) based on the dual-signal amplification of Hybridization Chain Reaction (HCR) and glucose oxidase (GOx) using silver coated glass slide (SCGS) as separation material.


Biomolecules detection Silver coated glass slide 3-(p-Hydroxyphenyl)-propanoic acid Horseradish peroxidase DNase I 



This work is supported by the Innovation Strong School Project of Guangdong education department (No. Q18291), the Non-funded Scientific and Technological Research Projects in Zhanjiang City (No. 2018B01005) and the program for scientific research start-up funds of Guangdong Ocean University (No. R17013).

Compliance with ethical standards

All the experiments are carried out following the relevant laws and institutional guidelines, and ethical standards.

Supplementary material

604_2019_3285_MOESM1_ESM.docx (20.9 mb)
ESM 1 (DOCX 21371 kb)


  1. 1.
    Becker-André M, Hahlbrock K (1989) Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by a PCR aided transcipt titration assay (PATTY). Nucleic Acids Res 17(17):9437–9446CrossRefGoogle Scholar
  2. 2.
    Xiang Y, Xie M, Bash R, Chen JL, Wang J (2007) Ultrasensitive label-free aptamer-based electronic detection. Angew Chem Int Ed Eng 46:9054–9056CrossRefGoogle Scholar
  3. 3.
    Li YB, Li RM, Zou L, Zhang MJ, Ling LL (2017) Fluorometric determination of simian virus 40 based on strand displacement amplification and triplex DNA using a molecular beacon probe with a guanine-rich fragment of the stem region. Microchim Acta 184:557–562CrossRefGoogle Scholar
  4. 4.
    Li YB, Liu S, Zhao ZK, Zheng YE, Wang ZR (2017) Binding induced strand displacement amplification for homogeneous protein assay. Talanta 164:196–200CrossRefGoogle Scholar
  5. 5.
    Li MMA, Li F, Zhang Z, Zhang K, Kang DK, Ankrum JA, Le XC, Zhao W (2014) Rolling circle amplification: a versatile tool for chemical biology, materials science and medicine. Chem Soc Rev 43:3324–3341CrossRefGoogle Scholar
  6. 6.
    He P, Liu L, Qiao W, Zhang S (2014) Ultrasensitive detection of thrombin using surface plasmon resonance and quartz crystal microbalance sensors by aptamer-based rolling circle amplification and nanoparticle signal enhancement. Chem Commun 50(12):1481–1484CrossRefGoogle Scholar
  7. 7.
    Liu S, Wang C, Zhang C, Wang Y, Tang B (2013) Label-free and ultrasensitive electrochemical detection of nucleic acids based on autocatalytic and exonuclease III-assisted target recycling strategy. Anal Chem 85:2282–2288CrossRefGoogle Scholar
  8. 8.
    Jiang W, Tian D, Zhang L, Guo Q, Cui Y, Yang M (2017) Dual signal amplification strategy for amperometric aptasensing using hydroxyapatite nanoparticles. Application to the sensitive detection of the cancer biomarker platelet-derived growth factor BB. Microchim Acta 184(11):4375–4381CrossRefGoogle Scholar
  9. 9.
    Dirks RM, Pierce NA (2004) Triggered amplification by hybridization chain reaction. Proc Natl Acad Sci 101:15275–15278CrossRefGoogle Scholar
  10. 10.
    Venkataraman S, Dirks RM, Rothemund PWK, Winfree E (2007) An autonomous polymerization motor powered by DNA hybridization. Pierce NA Nat Nanotechnol 2:490–494CrossRefGoogle Scholar
  11. 11.
    Yang L, Liu CH, Ren W, Li ZP (2012) Graphene surface-anchored fluorescence sensor for sensitive detection of MicroRNA coupled with enzyme-free signal amplification of hybridization chain reaction. ACS Appl Mater Interfaces 4:6450–6453CrossRefGoogle Scholar
  12. 12.
    Chen L, Sha L, Qiu Y, Wang G, Jiang H, Zhang X (2015) An amplified electrochemical aptasensor based on hybridization chain reactions and catalysis of silver nanoclusters. Nanoscale 7:3300–3308CrossRefGoogle Scholar
  13. 13.
    Guo Q, Han JJ, Shan S, Liu DF, Wu SS, Xiong YH, Lai WH (2016) DNA-based hybridization chain reaction and biotin–streptavidin signal amplification for sensitive detection of Escherichia coli O157:H7 through ELISA. Biosens Bioelectron 86:990–995CrossRefGoogle Scholar
  14. 14.
    Li ZB, Miao XM, Xing K, Zhu AH, Ling LL (2015) Enhanced electrochemical recognition of double-stranded DNA by using hybridization chain reaction and positively charged gold nanoparticles. Biosens Bioelectron 74:687–690CrossRefGoogle Scholar
  15. 15.
    Wang WJ, Li JJ, Rui K, Gai PP, Zhang JR, Zhu JJ (2015) Sensitive electrochemical detection of telomerase activity using spherical nucleic acids gold nanoparticles triggered mimic-hybridization chain reaction enzyme-free dual signal amplification. Anal Chem 87:3019–3026CrossRefGoogle Scholar
  16. 16.
    Li BL, Jiang Y, Chen X, Ellington AD (2012) Probing spatial organization of DNA strands using enzyme-free hairpin assembly circuits. J Am Chem Soc 134:13918–13921CrossRefGoogle Scholar
  17. 17.
    Lu WT, Arumugam R, Senapati D (2010) Multifunctional oval-shaped gold-nanoparticle-based selective detection of breast cancer cells using simple colorimetric and highly sensitive two-photon scattering assay. ACS Nano 4:1739–1749CrossRefGoogle Scholar
  18. 18.
    Duan N, Wu S, Chen X (2013) Selection and characterization of aptamers against Salmonella typhimurium using whole-bacterium systemic evolution of ligands by exponential enrichment (SELEX). Agric Food Chem 61:3229–2234CrossRefGoogle Scholar
  19. 19.
    Wang L, Tan W (2006) Multicolor FRET silica nanoparticles by single wavelength excitation. Nano Lett 6:84–88CrossRefGoogle Scholar
  20. 20.
    Ma X, Jiang Y, Jia F, Yu Y, Chen J, Wang Z (2014) An aptamer-based electrochemical biosensor for the detection of Salmonella. J Microbiol Methods 98:94–98CrossRefGoogle Scholar
  21. 21.
    Lian S, Zhang P, Gong P (2012) A universal quantum dots-aptamer probe for efficient cancer detection and targeted imaging. J Nanosci Nanotechnol 12:7703–7708CrossRefGoogle Scholar
  22. 22.
    Li YB, Liu S, Ling LL (2018) Sensitive fluorescent sensor for recognition of HIV-1 dsDNA by using glucose oxidase and triplex DNA. J Anal Methods Chem 2018:1–8. Article ID 8298365Google Scholar
  23. 23.
    Li YB, Liu S, Deng QJ, Ling LL (2018) A sensitive colorimetric DNA biosensor for specific detection of the HBV gene based on silver-coated glass slide and G-quadruplex-hemin DNAzyme. J Med Virol 90:699–705CrossRefGoogle Scholar
  24. 24.
    Li YB, Ling LL (2015) Aptamer-based fluorescent solid-phase thrombin assay using a silver-coated glass substrate and signal amplification by glucose oxidase. Microchim Acta 182:1849–1854CrossRefGoogle Scholar
  25. 25.
    Li YB, Zhang H, Zhu HY, Ling LL (2015) A sensitive fluorescence method for sequence specific recognition of single-stranded DNA by using glucose oxidase. Anal Methods 7:5436–5440CrossRefGoogle Scholar
  26. 26.
    Braun G, Lee SL, Dante M, Nguyen T, Moskovits M, Reich N (2007) Surface-enhanced Raman spectroscopy for DNA detection by nanoparticle assembly onto smooth metal films. J Am Chem Soc 129:6378–6379CrossRefGoogle Scholar
  27. 27.
    Lee J, Jean AKR, Hurst SJ, Mirkin CA (2007) Silver nanoparticle-oligonucleotide conjugates based on DNA with triple cyclic disulfide moieties. Nano Lett 7:2112–2115CrossRefGoogle Scholar
  28. 28.
    Thompson DG, Enright A, Faulds K, Smith WE, Graham D (2008) Ultrasensitive DNA detection using oligonucleotide-silver nanoparticle conjugates. Anal Chem 80:2805–2810CrossRefGoogle Scholar
  29. 29.
    Qu L, Dai L (2005) Novel silver nanostructures from silver mirror reaction on reactive substrates. J Phys Chem B 109:13985–13990CrossRefGoogle Scholar
  30. 30.
    Zhou Y, Li M, Su B, Lu Q (2009) Superhydrophobic surface created by the silver mirror reaction and its drag-reduction effect on water. J Mater Chem 19:3301–3306CrossRefGoogle Scholar
  31. 31.
    Lau OW, Shao B (2000) Determination of glucose using a piezoelectric quartz crystal and the silver mirror reaction. Anal Chim Acta 407:17–21CrossRefGoogle Scholar
  32. 32.
    Fang M, Grant P, McShane M, Sukhorukov G, Golub V, Lvov Y (2002) Magnetic bio/nanoreactor with multilayer shells of glucose oxidase and inorganic nanoparticles. Langmuir 18:6338–6344CrossRefGoogle Scholar
  33. 33.
    Lorenzo L, Rica R, Álvarez-Puebla R, Liz-Marzán L, Stevens M (2012) Plasmonic nanosensors with inverse sensitivity by means of enzyme-guided crystal growth. Nat Mater 11:604–607CrossRefGoogle Scholar
  34. 34.
    Baur J, Gondran C, Holzinger M, Defrancq E, Perrot H, Cosnier S (2010) Label-free femtomolar detection of target DNA by impedimetric DNA sensor based on poly(pyrrole-nitrilotriacetic acid) film. Anal Chem 82:1066–1072CrossRefGoogle Scholar
  35. 35.
    Wang P, Song YH, Zhao YJ, Fan AP (2013) Hydroxylamine amplified gold nanoparticle-based aptameric system for the highly selective and sensitive detection of platelet-derived growth factor. Talanta 103:392–397CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.School of Chemistry and EnvironmentGuangdong Ocean UniversityZhanjiangPeople’s Republic of China

Personalised recommendations