Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8294598 | Biochemical and Biophysical Research Communications | 2018 | 10 Pages |
Abstract
Gold nanoparticle (AuNP) is considered to be the most stable metal nanoparticle having the ability to be functionalized with biomolecules. Recently, AuNP-based DNA detection methods captured the interest of researchers worldwide. Paratuberculosis or Johne's disease, a chronic gastroenteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP), was found to have negative effect in the livestock industry. In this study, AuNP-based probes were evaluated for the specific and sensitive detection of MAP DNA. AuNP-based probe was produced by functionalization of AuNPs with thiol-modified oligonucleotide and was confirmed by Fourier-Transform Infrared (FTIR) spectroscopy. UV-Vis spectroscopy and Scanning Electron Microscopy (SEM) were used to characterize AuNPs. DNA detection was done by hybridization of 10â¯Î¼L of DNA with 5â¯Î¼L of probe at 63â¯Â°C for 10â¯min and addition of 3â¯Î¼L salt solution. The method was specific to MAP with detection limit of 103â¯ng. UV-Vis and SEM showed dispersion and aggregation of the AuNPs for the positive and negative results, respectively, with no observed particle growth. This study therefore reports an AuNP-based probes which can be used for the specific and sensitive detection of MAP DNA.
Keywords
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Authors
Thenor Aristotile Charles S. Ganareal, Michelle M. Balbin, Juvy J. Monserate, Joel R. Salazar, Claro N. Mingala,