An RNA-Aptamer-Based Assay for the Detection and Analysis of Malachite Green and Leucomalachite Green Residues in Fish Tissue

A robust screening assay employing solid phase extraction (SPE) followed by a novel aptamer-based procedure is presented for the rapid detection and semiquantitation of the triphenylmethane dye, Malachite Green (MG) and its primary metabolite Leucomalachite Green (LMG) in fish tissue. To the authors...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2010-04, Vol.82 (7), p.2652-2660
Main Authors: Stead, Sara L, Ashwin, Helen, Johnston, Brian H, Dallas, Anne, Kazakov, Sergei A, Tarbin, Jonathan A, Sharman, Matthew, Kay, Jack, Keely, Brendan J
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Aptamers, Nucleotide - chemistry
Chemistry
Dyes
Electrophoresis, Polyacrylamide Gel - methods
Exact sciences and technology
Fish
Fishes - metabolism
Food Contamination - analysis
Phase transitions
Ribonucleic acid
RNA
RNA - chemistry
Rosaniline Dyes - analysis
Rosaniline Dyes - isolation & purification
Solid Phase Extraction
Solids
Tissues
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Summary:A robust screening assay employing solid phase extraction (SPE) followed by a novel aptamer-based procedure is presented for the rapid detection and semiquantitation of the triphenylmethane dye, Malachite Green (MG) and its primary metabolite Leucomalachite Green (LMG) in fish tissue. To the authors’ knowledge, this is the first reported use of an RNA aptamer for the development of a diagnostic assay for the detection of chemical residues in food. The aptamer based screening assay is found to be highly specific for MG; but has negligible affinity for the LMG metabolite. However, because the LMG metabolite is lipophilic and known to be highly persistent in tissues, an oxidation step has been incorporated within the sample cleanup procedure to ensure that all LMG residues are converted to MG prior to measurement. This article provides evidence that an oligonucleotide aptamer can be used as an alternative recognition element to conventional antibodies with application to the detection of residues in food. Furthermore, this finding has the future potential to reduce the number of animals currently being used in the production of antibodies for immunodiagnostic kits.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac902226v