The effect of gold nanoparticles on the diagnostic polymerase chain reaction technique for equine herpes virus 1 (EHV-1)

Abstract

Nano-biotechnology has been a noticeable research area because of its successful applications in molecular diagnostics and therapy of various genetic and microbial diseases. Although the polymerase chain reaction (PCR) technique is one of the most highlighted and promising applications in the molecular diagnosis field, it suffers from some drawbacks that affect its efficiency. For instance, as a diagnostic technique for equine herpes virus-1 (EHV-1), conventional PCR could lead to false negative results due to the low viral titer in some samples, which leads to the necessity to improve its sensitivity. In this study, we carried out experiments to determine the effects of 15 nm unmodified citrate-coated gold nanoparticles (GNPs) on the key PCR reactants in order to see if these would enhance the overall outcomes of the reaction. Our results showed that, after optimization of the GNPs, oligonucleotide primers and Taq polymerase concentrations, a specific high yield amplification with a detection limit of 10² DNA copies could be reached compared to the 10⁵ to 10⁴ detection limit of conventional PCR. Thus, the developed and optimized GNPs-assisted PCR technique could be used for a more efficient, highly sensitive molecular detection of EHV-1.