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Nanoparticle sensor for label free detection of swine DNA in mixed biological samples

We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM...

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Main Authors: Ali, Eaqub, Hashim, Uda, Mustafa, Shuhaimi, Che Man, Yaakob, Yusop, Mohd Hazim, Bari, Md. Fazlul, Islam, Kh. Nurul, Hasan, M. F.
Format: Article
Language:English
Published: IOP Publishing 2011
Online Access:http://psasir.upm.edu.my/id/eprint/22268/1/Nanoparticle%20sensor%20for%20label%20free%20detection%20of%20swine%20DNA%20in%20mixed%20biological%20samples.pdf
http://psasir.upm.edu.my/id/eprint/22268/
http://iopscience.iop.org/article/10.1088/0957-4484/22/19/195503/meta;jsessionid=6FB4B035B384A6724959BDC78A24D781.ip-10-40-2-120
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spelling my.upm.eprints.222682017-08-25T10:13:52Z http://psasir.upm.edu.my/id/eprint/22268/ Nanoparticle sensor for label free detection of swine DNA in mixed biological samples Ali, Eaqub Hashim, Uda Mustafa, Shuhaimi Che Man, Yaakob Yusop, Mohd Hazim Bari, Md. Fazlul Islam, Kh. Nurul Hasan, M. F. We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530 nm and the appearance of new features in the 620–800 nm regions in their absorption spectra. The particles were stabilized against salt-induced aggregation upon the adsorption of single-stranded DNA. The PCR products, without any additional processing, were hybridized with a 17-base probe prior to exposure to GNPs. At a critical annealing temperature (55 °C) that differentiated matched and mismatched base pairing, the probe was hybridized to pig PCR product and dehybridized from the deer product. The dehybridized probe stuck to GNPs to prevent them from salt-induced aggregation and retained their characteristic red color. Hybridization of a 27-nucleotide probe to swine mitochondrial DNA identified them in pork–venison, pork–shad and venison–shad binary admixtures, eliminating the need of PCR amplification. Thus the assay was applied to authenticate species both in PCR-amplified and non-amplified heterogeneous biological samples. The results were determined visually and validated by absorption spectroscopy. The entire assay (hybridization plus visual detection) was performed in less than 10 min. The LOD (for genomic DNA) of the assay was 6 µg ml − 1 swine DNA in mixed meat samples. We believe the assay can be applied for species assignment in food analysis, mismatch detection in genetic screening and homology studies between closely related species. IOP Publishing 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22268/1/Nanoparticle%20sensor%20for%20label%20free%20detection%20of%20swine%20DNA%20in%20mixed%20biological%20samples.pdf Ali, Eaqub and Hashim, Uda and Mustafa, Shuhaimi and Che Man, Yaakob and Yusop, Mohd Hazim and Bari, Md. Fazlul and Islam, Kh. Nurul and Hasan, M. F. (2011) Nanoparticle sensor for label free detection of swine DNA in mixed biological samples. Nanotechnology, 22 (19). pp. 1-8. ISSN 0957-4484; ESSN: 1361-6528 http://iopscience.iop.org/article/10.1088/0957-4484/22/19/195503/meta;jsessionid=6FB4B035B384A6724959BDC78A24D781.ip-10-40-2-120 10.1088/0957-4484/22/19/195503
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description We used 40 ± 5 nm gold nanoparticles (GNPs) as colorimetric sensor to visually detect swine-specific conserved sequence and nucleotide mismatch in PCR-amplified and non-amplified mitochondrial DNA mixtures to authenticate species. Colloidal GNPs changed color from pinkish-red to gray-purple in 2 mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530 nm and the appearance of new features in the 620–800 nm regions in their absorption spectra. The particles were stabilized against salt-induced aggregation upon the adsorption of single-stranded DNA. The PCR products, without any additional processing, were hybridized with a 17-base probe prior to exposure to GNPs. At a critical annealing temperature (55 °C) that differentiated matched and mismatched base pairing, the probe was hybridized to pig PCR product and dehybridized from the deer product. The dehybridized probe stuck to GNPs to prevent them from salt-induced aggregation and retained their characteristic red color. Hybridization of a 27-nucleotide probe to swine mitochondrial DNA identified them in pork–venison, pork–shad and venison–shad binary admixtures, eliminating the need of PCR amplification. Thus the assay was applied to authenticate species both in PCR-amplified and non-amplified heterogeneous biological samples. The results were determined visually and validated by absorption spectroscopy. The entire assay (hybridization plus visual detection) was performed in less than 10 min. The LOD (for genomic DNA) of the assay was 6 µg ml − 1 swine DNA in mixed meat samples. We believe the assay can be applied for species assignment in food analysis, mismatch detection in genetic screening and homology studies between closely related species.
format Article
author Ali, Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Yusop, Mohd Hazim
Bari, Md. Fazlul
Islam, Kh. Nurul
Hasan, M. F.
spellingShingle Ali, Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Yusop, Mohd Hazim
Bari, Md. Fazlul
Islam, Kh. Nurul
Hasan, M. F.
Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
author_facet Ali, Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Yusop, Mohd Hazim
Bari, Md. Fazlul
Islam, Kh. Nurul
Hasan, M. F.
author_sort Ali, Eaqub
title Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
title_short Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
title_full Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
title_fullStr Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
title_full_unstemmed Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
title_sort nanoparticle sensor for label free detection of swine dna in mixed biological samples
publisher IOP Publishing
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/22268/1/Nanoparticle%20sensor%20for%20label%20free%20detection%20of%20swine%20DNA%20in%20mixed%20biological%20samples.pdf
http://psasir.upm.edu.my/id/eprint/22268/
http://iopscience.iop.org/article/10.1088/0957-4484/22/19/195503/meta;jsessionid=6FB4B035B384A6724959BDC78A24D781.ip-10-40-2-120
_version_ 1643827778391375872
score 13.211869

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