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dc.contributor.authorSaikrishnan, Deepika
dc.contributor.authorGoyal, Madhu
dc.contributor.authorRossiter, Sharon
dc.contributor.authorKukol, A.
dc.date.accessioned2014-12-11T11:47:26Z
dc.date.available2014-12-11T11:47:26Z
dc.date.issued2014-12-01
dc.identifier.citationSaikrishnan , D , Goyal , M , Rossiter , S & Kukol , A 2014 , ' A cellulose-based bioassay for the colorimetric detection of pathogen DNA ' , Analytical and Bioanalytical Chemistry , vol. 406 , no. 30 , pp. 7887-7898 . https://doi.org/10.1007/s00216-014-8257-y
dc.identifier.issn1618-2642
dc.identifier.otherPURE: 7848520
dc.identifier.otherPURE UUID: 0c3f7603-d0d4-44da-b3f0-6e9d77cf6616
dc.identifier.otherScopus: 84913580413
dc.identifier.urihttp://hdl.handle.net/2299/14930
dc.description.abstractCellulose-paper-based colorimetric bioassays may be used at the point of sampling without sophisticated equipment. This study reports the development of a colorimetric bioassay based on cellulose that can detect pathogen DNA. The detection was based on covalently attached single-stranded DNA probes and visual analysis. A cellulose surface functionalized with tosyl groups was prepared by the N,N-dimethylacetamide-lithium chloride method. Tosylation of cellulose was confirmed by scanning electron microscopy, Fourier transform infrared spectroscopy and elemental analysis. Sulfhydryl-modified oligonucleotide probes complementary to a segment of the DNA sequence IS6110 of Mycobacterium tuberculosis were covalently immobilized on the tosylated cellulose. On hybridization of biotin-labelled DNA oligonucleotides with these probes, a colorimetric signal was obtained with streptavidin-conjugated horseradish peroxidase catalysing the oxidation of tetramethylbenzamidine by H2O2. The colour intensity was significantly reduced when the bioassay was subjected to DNA oligonucleotide of randomized base composition. Initial experiments have shown a sensitivity of 0.1 μM. A high probe immobilization efficiency (more than 90 %) was observed with a detection limit of 0.1 μM, corresponding to an absolute amount of 10 pmol. The detection of M. tuberculosis DNA was demonstrated using this technique coupled with PCR for biotinylation of the DNA. This work shows the potential use of tosylated cellulose as the basis for point-of-sampling bioassays.en
dc.language.isoeng
dc.relation.ispartofAnalytical and Bioanalytical Chemistry
dc.subjectBiosensor
dc.subjectcellulose
dc.subjecttuberculosis
dc.subjectOligonucleotides, Antisense
dc.titleA cellulose-based bioassay for the colorimetric detection of pathogen DNAen
dc.contributor.institutionSchool of Life and Medical Sciences
dc.contributor.institutionDepartment of Human and Environmental Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.contributor.institutionVeterinary Science
dc.contributor.institutionMicrobiology and Biotechnology
dc.contributor.institutionGeography, Environment and Agriculture
dc.contributor.institutionAgriculture, Food and Veterinary Sciences
dc.contributor.institutionDepartment of Pharmacy
dc.contributor.institutionMedicinal and Analytical Chemistry
dc.contributor.institutionNatural Product Chemistry and Drug Design
dc.contributor.institutionPharmacology and Clinical Science Research
dc.contributor.institutionCardiovascular Pathologies
dc.contributor.institutionBiochemistry and Bioinformatics
dc.contributor.institutionBiosciences Research Group
dc.description.statusPeer reviewed
dc.date.embargoedUntil2015-10-01
rioxxterms.versionAM
rioxxterms.versionAM
rioxxterms.versionofrecordhttps://doi.org/10.1007/s00216-014-8257-y
rioxxterms.typeJournal Article/Review
herts.preservation.rarelyaccessedtrue


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