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Int J Biol Macromol ; 124: 1256-1263, 2019 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-30445082

RESUMEN

S. aureus is one of important causes of disease, food poisoning in humans and animals. The generally methods for detection of S. aureus is time consuming. Therefore, a new method is necessary for rapid, sensitive and specific diagnosis of S. aureus. In the present study, two probes and a Bio-barcode DNA were designed for detection of S. aureus (Protein A). Firstly, magnetic nanoparticle (MNPs) and gold nanoparticle (AuNPs) were synthesized at 80 °C and 100 °C, respectively. The AuNPs and the MNPs were functionalized with probe1, Bio-barcode DNA and probe2, respectively. Target DNA was added into the nanomaterial's system containing bio-barcode DNA-AuNPs-probe1 and probe2-MNPs to formed bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex. The bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex was separated with magnetic field. Finally, the bio-barcode DNA was released from surface of complex using DTT (0.8 M) and there was isolated of nanoparticles by magnetic field and centrifuge. The fluorescence intensity of bio-barcode DNA was measured in different concentrations of S. aureus (101 to 108 CFU mL-1) by fluorescence spectrophotometry. The results showed that standard curve was linearly from 102 to 107 CFU mL-1. Limit of detection of bio-barcode assay for both PBS and real samples was 86 CFU mL-1.


Asunto(s)
Técnicas de Tipificación Bacteriana , Técnicas Biosensibles , Código de Barras del ADN Taxonómico/métodos , ADN Bacteriano/química , Nanopartículas del Metal/química , Proteína Estafilocócica A/análisis , Staphylococcus aureus/genética , Sondas de ADN/síntesis química , Sondas de ADN/química , ADN Bacteriano/metabolismo , Oro/química , Humanos , Hierro/química , Límite de Detección , Imanes , Espectrometría de Fluorescencia , Staphylococcus aureus/clasificación , Staphylococcus aureus/aislamiento & purificación
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