tetano
Editor, Senior Moderator
Mikrochim Acta. 2019 Feb 2;186(3):155. doi: 10.1007/s00604-019-3285-0.
[h=1]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.[/h] Li Y[SUP]1[/SUP], Shao J[SUP]2[/SUP], Guo W[SUP]2[/SUP], Wang M[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 H[SUB]2[/SUB]O[SUB]2[/SUB]. The H[SUB]2[/SUB]O[SUB]2[/SUB] 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[SUP]-1[/SUP] PDGF-BB concentration range, and the detection limit is 191 fmol?L[SUP]-1[/SUP]. For the H1N1 virus DNA, the respective data are 2.5 to 300 pmol?L[SUP]-1[/SUP] and 826 fmol?L[SUP]-1[/SUP]. 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.
[h=4]KEYWORDS:[/h] 3-(p-Hydroxyphenyl)-propanoic acid; Biomolecules detection; DNase I; Horseradish peroxidase; Silver coated glass slide
PMID: 30712102 DOI: 10.1007/s00604-019-3285-0
[h=1]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.[/h] Li Y[SUP]1[/SUP], Shao J[SUP]2[/SUP], Guo W[SUP]2[/SUP], Wang M[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 H[SUB]2[/SUB]O[SUB]2[/SUB]. The H[SUB]2[/SUB]O[SUB]2[/SUB] 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[SUP]-1[/SUP] PDGF-BB concentration range, and the detection limit is 191 fmol?L[SUP]-1[/SUP]. For the H1N1 virus DNA, the respective data are 2.5 to 300 pmol?L[SUP]-1[/SUP] and 826 fmol?L[SUP]-1[/SUP]. 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.
[h=4]KEYWORDS:[/h] 3-(p-Hydroxyphenyl)-propanoic acid; Biomolecules detection; DNase I; Horseradish peroxidase; Silver coated glass slide
PMID: 30712102 DOI: 10.1007/s00604-019-3285-0