Multiplex Nanoparticle-Assisted PCR (NanoPCR) for Detection of Clinically Important Bacterial and Fungal Pathogens

Nano-PCR is a technique that uses the physicochemical properties

of nanomaterials in order to enhance specificity and sensitivity of PCR.

Several studies showed the potential application of nanoparticles (NPs)

for detection of bacterial, parasitic, and viral infections. However, to

our knowledge, NPs have not been used to detect fungal and bacterial

pathogens in a single PCR. Here, we aimed to develop and optimize a

novel multiplex nano-PCR assay for detection of health-threatening

fungal and bacterial strains. For this bpurpose, conserved regions in the

genomes of pathogens were amplified using the species-spesific primers

by multiplex PCR. After confirmation of PCR products by agarose gel

electrophoresis, multiplex nano-PCR was performed in the presence of

gold NPs, quantum dots, and metal NPs. The effect of each nanoparticle

on PCR was evaluated by comparing the PCR cycle number, limit of

detection, and amplification efficiency. In conclusion, we developed

an effective tool serving as spesific, sensitive and faster alternative to

conventional pathogen detection methods