Development of nanoparticle-assisted PCR assay in the rapid detection of brain-eating amoebae

Gabriel, Shobana * and Rasheed, A. K. * and Siddiqui, Ruqaiyyah Bano * and Appaturi, Jimmy Nelson and Leo, Bey Fen (2018) Development of nanoparticle-assisted PCR assay in the rapid detection of brain-eating amoebae. Parasitology Research, 117 (6). pp. 1801-1811. ISSN 0023-6438

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Official URL: https://link.springer.com/article/10.1007%2Fs00436...

Abstract

Brain-eating amoebae (Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri) have gained increasing attention owing to their capacity to produce severe human and animal infections involving the brain. Early detection is a pre-requisite in successful prognosis. Here, we developed a nanoPCR assay for the rapid detection of brain-eating amoebae using various nanoparticles. Graphene oxide, copper and alumina nanoparticles used in this study were characterized using Raman spectroscopy measurements through excitation with a He–Ne laser, while powder X-ray diffraction patterns were taken on a PANanalytical, X’Pert HighScore diffractometer and the morphology of the materials was confirmed using high-resolution transmission electron microscopy (HRTEM). Using nanoparticle-assisted PCR, the results revealed that graphene oxide, copper oxide and alumina nanoparticles significantly enhanced PCR efficiency in the detection of pathogenic free-living amoebae using genus-specific probes. The optimal concentration of graphene oxide, copper oxide and alumina nanoparticles for Acanthamoeba spp. was determined at 0.4, 0.04 and 0.4 μg per mL respectively. For B. mandrillaris, the optimal concentration was determined at 0.4 μg per mL for graphene oxide, copper oxide and alumina nanoparticles, and for Naegleria, the optimal concentration was 0.04, 4.0 and 0.04 μg per mL respectively. Moreover, combinations of these nanoparticles proved to further enhance PCR efficiency. The addition of metal oxide nanoparticles leads to excellent surface effect, while thermal conductivity property of the nanoparticles enhances PCR productivity. These findings suggest that nanoPCR assay has tremendous potential in the clinical diagnosis of parasitic infections as well as for studying epidemiology and pathology and environmental monitoring of other microbes.

Item Type: Article
Uncontrolled Keywords: Acanthamoeba spp; Balamuthia mandrillaris; Naegleria; PCR; detection; nanoparticles
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Sunway University > School of Engineering and Technology [formerly School of Science and Technology until 2020] > Dept. Biological Sciences moved to SMLS wef 2021
Sunway University > School of Engineering and Technology [formerly School of Science and Technology until 2020] > Research Centre for Nano-Materials and Energy Technology
Depositing User: Dr Janaki Sinnasamy
Related URLs:
Date Deposited: 10 Dec 2018 08:54
Last Modified: 30 Apr 2019 08:45
URI: http://eprints.sunway.edu.my/id/eprint/981

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