Nanoparticle sensor for label free detection of swine DNA in mixed biological samples
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2011
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my.unimap-137912012-05-03T05:02:53Z Nanoparticle sensor for label free detection of swine DNA in mixed biological samples M. E., Ali Uda, Hashim, Prof. Dr. S., Mustafa Y. B., Che Man M. H. M., Yusop Mohd Fazalul, Bari Kh. N., Islam M. F., Hasan uda@unimap.edu.my Gold nanoparticles (GNPs) Swine DNA Nanoparticle sensor Label-free detection Link to publisher's homepage at http://iopscience.iop.org/ We used 40 5nm 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 2mM PBS. Visually observed results were clearly reflected by the dramatic reduction of surface plasmon resonance peak at 530nm and the appearance of new features in the 620-800nm 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 10min. The LOD (for genomic DNA) of the assay was 6νgml-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. 2011-09-19T03:01:47Z 2011-09-19T03:01:47Z 2011-05-13 Article Nanotechnology, vol. 22(19), 2011 0957-4484 http://iopscience.iop.org/0957-4484/22/19/195503/pdf/0957-4484_22_19_195503.pdf http://hdl.handle.net/123456789/13791 10.1088/0957-4484/22/19/195503 en IOP Publishing Ltd. |
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English |
| topic |
Gold nanoparticles (GNPs) Swine DNA Nanoparticle sensor Label-free detection |
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Gold nanoparticles (GNPs) Swine DNA Nanoparticle sensor Label-free detection M. E., Ali Uda, Hashim, Prof. Dr. S., Mustafa Y. B., Che Man M. H. M., Yusop Mohd Fazalul, Bari Kh. N., Islam M. F., Hasan Nanoparticle sensor for label free detection of swine DNA in mixed biological samples |
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Link to publisher's homepage at http://iopscience.iop.org/ |
| author2 |
uda@unimap.edu.my |
| author_facet |
uda@unimap.edu.my M. E., Ali Uda, Hashim, Prof. Dr. S., Mustafa Y. B., Che Man M. H. M., Yusop Mohd Fazalul, Bari Kh. N., Islam M. F., Hasan |
| format |
Article |
| author |
M. E., Ali Uda, Hashim, Prof. Dr. S., Mustafa Y. B., Che Man M. H. M., Yusop Mohd Fazalul, Bari Kh. N., Islam M. F., Hasan |
| author_sort |
M. E., Ali |
| 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 Ltd. |
| publishDate |
2011 |
| url |
http://dspace.unimap.edu.my/xmlui/handle/123456789/13791 |
| _version_ |
1643791011799891968 |
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13.214268 |