Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats

A novel class of nanobiosensor was developed by integrating a 27-nucleotide AluI fragment of swine cytochrome b (cytb) gene to a 3-nm diameter citrate-tannate coated gold nanoparticle (GNP). The biosensor detected 0.5% and 1% pork in raw and 2.5-h autoclaved pork-beef binary admixtures in a single s...

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Main Authors: Ali, Md. Eaqub, Hashim, Uda, Mustafa, Shuhaimi, Che Man, Yaakob, Mohd Yusop, Mohd Hazim, Kashif, M., Dhahi, Thikra S., Bari, M. F., Hakim, M. A., Abdul Latif, Muhammad
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2011
Online Access:http://psasir.upm.edu.my/id/eprint/22311/
http://psasir.upm.edu.my/id/eprint/22311/1/22311.pdf
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author Ali, Md. Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Mohd Yusop, Mohd Hazim
Kashif, M.
Dhahi, Thikra S.
Bari, M. F.
Hakim, M. A.
Abdul Latif, Muhammad
author_facet Ali, Md. Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Mohd Yusop, Mohd Hazim
Kashif, M.
Dhahi, Thikra S.
Bari, M. F.
Hakim, M. A.
Abdul Latif, Muhammad
author_sort Ali, Md. Eaqub
building UPM Institutional Repository
collection Online Access
description A novel class of nanobiosensor was developed by integrating a 27-nucleotide AluI fragment of swine cytochrome b (cytb) gene to a 3-nm diameter citrate-tannate coated gold nanoparticle (GNP). The biosensor detected 0.5% and 1% pork in raw and 2.5-h autoclaved pork-beef binary admixtures in a single step without any separation or washing. The hybridization kinetics of the hybrid sensor was studied with synthetic and AluI digested real pork targets from moderate to extreme target concentrations and a sigmoidal relationship was found. Using the kinetic curve, a convenient method for quantifying and counting target DNA copy number was developed. The accuracy of the method was over 90% and 80% for raw and autoclaved pork-beef binary admixtures in the range of 5–100% pork adulteration. The biosensor probe identified a target DNA sequence that was several-folds shorter than a typical PCR-template. This offered the detection and quantitation of potential targets in highly processed or degraded samples where PCR amplification was not possible due to template crisis. The assay was a viable alternative approach of qPCR for detecting, quantifying and counting copy number of shorter size DNA sequences to address a wide ranging biological problem in food industry, diagnostic laboratories and forensic medicine.
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spelling upm-223112019-12-18T04:32:13Z http://psasir.upm.edu.my/id/eprint/22311/ Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats Ali, Md. Eaqub Hashim, Uda Mustafa, Shuhaimi Che Man, Yaakob Mohd Yusop, Mohd Hazim Kashif, M. Dhahi, Thikra S. Bari, M. F. Hakim, M. A. Abdul Latif, Muhammad A novel class of nanobiosensor was developed by integrating a 27-nucleotide AluI fragment of swine cytochrome b (cytb) gene to a 3-nm diameter citrate-tannate coated gold nanoparticle (GNP). The biosensor detected 0.5% and 1% pork in raw and 2.5-h autoclaved pork-beef binary admixtures in a single step without any separation or washing. The hybridization kinetics of the hybrid sensor was studied with synthetic and AluI digested real pork targets from moderate to extreme target concentrations and a sigmoidal relationship was found. Using the kinetic curve, a convenient method for quantifying and counting target DNA copy number was developed. The accuracy of the method was over 90% and 80% for raw and autoclaved pork-beef binary admixtures in the range of 5–100% pork adulteration. The biosensor probe identified a target DNA sequence that was several-folds shorter than a typical PCR-template. This offered the detection and quantitation of potential targets in highly processed or degraded samples where PCR amplification was not possible due to template crisis. The assay was a viable alternative approach of qPCR for detecting, quantifying and counting copy number of shorter size DNA sequences to address a wide ranging biological problem in food industry, diagnostic laboratories and forensic medicine. Hindawi Publishing Corporation 2011 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/22311/1/22311.pdf Ali, Md. Eaqub and Hashim, Uda and Mustafa, Shuhaimi and Che Man, Yaakob and Mohd Yusop, Mohd Hazim and Kashif, M. and Dhahi, Thikra S. and Bari, M. F. and Hakim, M. A. and Abdul Latif, Muhammad (2011) Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats. Journal of Nanomaterials, 2011. art. no. 781098. pp. 1-11. ISSN 1687-4110; ESSN: 1687-4129 https://www.hindawi.com/journals/jnm/2011/781098/ 10.1155/2011/781098
spellingShingle Ali, Md. Eaqub
Hashim, Uda
Mustafa, Shuhaimi
Che Man, Yaakob
Mohd Yusop, Mohd Hazim
Kashif, M.
Dhahi, Thikra S.
Bari, M. F.
Hakim, M. A.
Abdul Latif, Muhammad
Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title_full Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title_fullStr Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title_full_unstemmed Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title_short Nanobiosensor for detection and quantification of DNA sequences in degraded mixed meats
title_sort nanobiosensor for detection and quantification of dna sequences in degraded mixed meats
url http://psasir.upm.edu.my/id/eprint/22311/
http://psasir.upm.edu.my/id/eprint/22311/
http://psasir.upm.edu.my/id/eprint/22311/
http://psasir.upm.edu.my/id/eprint/22311/1/22311.pdf