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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
  • 14 Downloads

Abstract

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.

Keywords

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

Notes

Acknowledgments

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)

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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

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