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Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.

A new electrochemical biosensor is described for voltammetric detection of gene sequence related to Trichoderma harzianum. The sensor involves immobilization of a 20 base single-stranded probe (ssDNA), which is complementary to a specific gene sequence related to T. harzianum on a gold electrode thr...

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Main Authors: Siddiquee, Shafiquzzaman, Yusof, Nor Azah, Salleh, Abu Bakar, Abu Bakar, Fatimah, Lee, Yook Heng
Format: Article
Language:English
Published: Elsevier 2010
Online Access:http://psasir.upm.edu.my/id/eprint/14462/
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spelling my.upm.eprints.144622014-02-12T07:57:05Z http://psasir.upm.edu.my/id/eprint/14462/ Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species. Siddiquee, Shafiquzzaman Yusof, Nor Azah Salleh, Abu Bakar Abu Bakar, Fatimah Lee, Yook Heng A new electrochemical biosensor is described for voltammetric detection of gene sequence related to Trichoderma harzianum. The sensor involves immobilization of a 20 base single-stranded probe (ssDNA), which is complementary to a specific gene sequence related to T. harzianum on a gold electrode through specific adsorption. The DNA probe was used to determine the amount of target gene in solution using methylene blue (MB) as the electrochemical indicator. The covalently immobilized probe could selectively hybridize with the target DNA to form a hybrid on the surface despite the bases being attached to the electrode. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with the target. Peak currents were found to increase in the following order: hybrid-modified AuE and the probe-modified AuE which localized to the affinity of MB. Control experiments with the non-complementary oligonucleotides were performed to assess whether the DNA biosensor responds selectively, via hybridization, to the target. DNA biosensor also able to detect microorganism at the species levels without nucleic acid amplification. The redox current was linearly related to the concentration of target oligonucleotide DNA, ranged from 1–20 ppm. Numerous factors, affecting the probe immobilization, target hybridization and indicator binding reactions are optimized to maximize the sensitivity and reduce the assay time. Elsevier 2010-08 Article PeerReviewed Siddiquee, Shafiquzzaman and Yusof, Nor Azah and Salleh, Abu Bakar and Abu Bakar, Fatimah and Lee, Yook Heng (2010) Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species. Bioelectrochemistry, 79 (1). pp. 31-36. ISSN 1567-5394; ESSN: 1878-562X 10.1016/j.bioelechem.2009.10.004 English
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 A new electrochemical biosensor is described for voltammetric detection of gene sequence related to Trichoderma harzianum. The sensor involves immobilization of a 20 base single-stranded probe (ssDNA), which is complementary to a specific gene sequence related to T. harzianum on a gold electrode through specific adsorption. The DNA probe was used to determine the amount of target gene in solution using methylene blue (MB) as the electrochemical indicator. The covalently immobilized probe could selectively hybridize with the target DNA to form a hybrid on the surface despite the bases being attached to the electrode. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with the target. Peak currents were found to increase in the following order: hybrid-modified AuE and the probe-modified AuE which localized to the affinity of MB. Control experiments with the non-complementary oligonucleotides were performed to assess whether the DNA biosensor responds selectively, via hybridization, to the target. DNA biosensor also able to detect microorganism at the species levels without nucleic acid amplification. The redox current was linearly related to the concentration of target oligonucleotide DNA, ranged from 1–20 ppm. Numerous factors, affecting the probe immobilization, target hybridization and indicator binding reactions are optimized to maximize the sensitivity and reduce the assay time.
format Article
author Siddiquee, Shafiquzzaman
Yusof, Nor Azah
Salleh, Abu Bakar
Abu Bakar, Fatimah
Lee, Yook Heng
spellingShingle Siddiquee, Shafiquzzaman
Yusof, Nor Azah
Salleh, Abu Bakar
Abu Bakar, Fatimah
Lee, Yook Heng
Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
author_facet Siddiquee, Shafiquzzaman
Yusof, Nor Azah
Salleh, Abu Bakar
Abu Bakar, Fatimah
Lee, Yook Heng
author_sort Siddiquee, Shafiquzzaman
title Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
title_short Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
title_full Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
title_fullStr Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
title_full_unstemmed Electrochemical DNA biosensor for the detection of specific gene related to Trichoderma harzianum species.
title_sort electrochemical dna biosensor for the detection of specific gene related to trichoderma harzianum species.
publisher Elsevier
publishDate 2010
url http://psasir.upm.edu.my/id/eprint/14462/
_version_ 1643825639924432896
score 13.211869

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