• 文献标题:   A fluorescence quenching based gene assay for Escherichia coli O157:H7 using graphene quantum dots and gold nanoparticles
  • 文献类型:   Article
  • 作  者:   SAAD SM, ABDULLAH J, ABD RASHID S, FEN YW, SALAM F, YIH LH
  • 作者关键词:   fluorophore, quencher, food pathogen, flic gene, static quenching, real sample, selectivity
  • 出版物名称:   MICROCHIMICA ACTA
  • ISSN:   0026-3672 EI 1436-5073
  • 通讯作者地址:   Univ Putra Malaysia
  • 被引频次:   2
  • DOI:   10.1007/s00604-019-3913-8
  • 出版年:   2019

▎ 摘  要

A fluorometric assay is described for highly sensitive quantification of Escherichia coli O157:H7. Reporter oligos were immobilized on graphene quantum dots (GQDs), and quencher oligos were immobilized on gold nanoparticles (AuNPs). Target DNA was co-hybridized with reporter oligos on the GQDs and quencher oligos on AuNPs. This triggers quenching of fluorescence (with excitation/emission peaks at 400 nm/530 nm). On introducing target into the system, fluorescence is quenched by up to 95% by 100 nM concentrations of target oligos having 20 bp. The response to the fliC gene of E. coli O157:H7 increases with the logarithm of the concentration in the range from 0.1 nM to 150 nM. The limit of detection is 1.1 +/- 0.6 nM for n = 3. The selectivity and specificity of the assay was confirmed by evaluating the various oligos sequences and PCR product (fliC gene) amplified from genomic DNA of the food samples spiked with E. coli O157:H7.