The genus Trichoderma is a soil-borne fungi which in numerous reports has been successfully used as a
biological control agent against various plant pathogens. The identification of Trichoderma species worldwide
is currently deduced from micro-morphological descriptions which are tedious and prone to error.
Electrochemical approaches are currently being developed for the detection and analysis of DNA. In the
present study, an electrochemical DNA biosensor was successfully developed based on ionic liquid (e.g.,
1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM][Otf])), ZnO nanoparticles and a chitosan
(CHIT) nanocomposite membrane on a modified gold electrode (AuE). A single-stranded DNA probe
was immobilized on this electrode. Methylene blue (MB) was used as the hybridization indicator to
monitor the hybridization reaction of the target DNA. Under optimal conditions using differential
pulse voltammetry (DPV), the target DNA sequences were detectable at concentration ranges of
1.0 1018–1.82 104 mol L1, and the detectable limit was 1.0 1019 mol L1. The developed DNA
biosensor enables the study of hybridization with crude DNA fragments and the results of this study
confirm that this DNA biosensor provides a fast, sensitive and convenient way for the species level
identification of Trichoderma harzianum.
2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license
The genus Trichoderma is a soil-borne fungi which in numerous reports has been successfully used as a
biological control agent against various plant pathogens. The identification of Trichoderma species worldwide
is currently deduced from micro-morphological descriptions which are tedious and prone to error.
Electrochemical approaches are currently being developed for the detection and analysis of DNA. In the
present study, an electrochemical DNA biosensor was successfully developed based on ionic liquid (e.g.,
1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM][Otf])), ZnO nanoparticles and a chitosan
(CHIT) nanocomposite membrane on a modified gold electrode (AuE). A single-stranded DNA probe
was immobilized on this electrode. Methylene blue (MB) was used as the hybridization indicator to
monitor the hybridization reaction of the target DNA. Under optimal conditions using differential
pulse voltammetry (DPV), the target DNA sequences were detectable at concentration ranges of
1.0 1018–1.82 104 mol L1, and the detectable limit was 1.0 1019 mol L1. The developed DNA
biosensor enables the study of hybridization with crude DNA fragments and the results of this study
confirm that this DNA biosensor provides a fast, sensitive and convenient way for the species level
identification of Trichoderma harzianum.
2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license
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