Nanoparticle-assisted Polymerase Chain Reaction (NanoPCR): Optimization of PCR detection of Leifsonia xyli subsp. xyli by the addition of nanoparticles

Abstract

Leifsonia xyli subsp. xyli (Lxx) causes ratoon stunting disease (RSD) in sugarcane, and is one of major causes of production losses. The detection of Lxx bacteria in sugarcane is made mainly through molecular biology techniques, especially polymerase chain reaction (PCR). However, PCR presents some barriers to provide reliable results. The present work brings a Nanoparticle-assisted Polymerase Chain Reaction (NanoPCR) assay for the detection of Lxx in its latent infection on micropropagated sugarcane. This assay was based in the addition of Gold and Titanium dioxide nanoparticles to conventional PCR and evaluation of its effects. It was observed that the reactions performed with Titanium dioxide nanoparticles provided the formation of singular well-defined bands under electrophoresis, consistent with the expected molecular weight, without occurrence of non-specific bands or presenting false negatives occurrence, negative effects that were observed in the control assay. While the performed NanoPCR adding AuNP also provided the formation of well-defined bands, been able to inhibit the occurrence of false negatives, but wasn’t able to eliminate the occurrence of non-specific amplifications. The results indicate that NanoPCR by the addition of Gold and Titanium dioxide nanoparticles to conventional PCR increased the detection of Lxx.