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Published on: June 23, 2016
Development of a Gold Nanoparticle-Assisted Multiplex Polymerase Chain Reaction Assay for Selected Hospital-Acquired
Kubra Aslan1,2, Emre Erden Kopar2, Serap Evran2
1Chemistry Department, Faculty of Science, Ataturk University, Yakutiye, Erzurum, Türkiye.
Abstract:
In this study, development and optimization of a multiplex PCR assay for the simultaneous amplification of selected bacterial and fungal targets were performed. Source of hospital-acquired infections-specific, (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Candida albicans, Fusarium oxysporum, and Scedosporium prolificans) primer sets targeting conserved genomic regions were systematically evaluated for potential nonspecific interactions, cross-reactivity, and structural stability using in silico tools and empirical analyses. Multiplex PCR conditions were optimized using genomic DNA from reference strains under controlled laboratory conditions, with particular emphasis on annealing temperature, MgCl2 concentration, and primer balance. In addition, the effect of gold nanoparticles (AuNPs) on amplification performance was investigated. The inclusion of 1.0 nM 10 nm AuNPs enhanced overall amplification efficiency, with a more pronounced improvement observed for fungal targets. Overall, this study establishes an analytical proof of concept for a nanoparticle-assisted multiplex PCR configuration capable of simultaneously amplifying multiple bacterial and fungal targets in a single reaction.
