DEVELOPMENT OF PCR BASED RAPID ASSAY METHOD FOR THE DETECTION OF AFLATOXIN IN FOODS USING 2100 BIOANALYZERS. JANAKI ALIAS PRIYA, ANURAG CHATURVEDI , MANORAMA KANURI, RAVICHARAN AMEDA , PRATHIMA NAMBURI , SREEDHAR MULINTI,M.R. KRISHNA BHAGAVATULA AND SUNIL CHIDAMBAR KULKARNI ANURAG CHATHURVEDI , MANORAMA KANURIA, PRATHIMA NAMBURIB, M.R. KRISHNA BHAGAVATUAbstract Aflatoxins are toxic and carcinogenic polyketide metabolities produced by the species Aspergillus flavus, Aspergillus Parasiticus. Contamination of foods and animal feeds with aflatoxin is a worldwide problem. Aflatoxin contamination constitutes one of the major health hazard groups of naturally occurring toxicants both for humans and animals. The present study was undertaken with the main aim of developing rapid PCR based method for detection of aflatoxin in foods. Aflatoxin producing gene fragments in fungal strains were amplified using PCR along with an Agilent 2100 Bioanalyzer in an attempt to develop a rapid assay for aflatoxin detection. Forward and reverse primers were designed from original gene sequences (obtained from GENBANK) of the aflatoxin genes, aflQ, which are key genes involved in the production of the Aflatoxin, using DNASTAR Lasergene 8.0 version software. These primers were used for amplification of a 166 base pair fragment of aflQ target from DNA of Aspergillus parasiticus and Aspergillus flavus. The fragments obtained were resolved on a DNA 1000 LabchipR in the Agilent 2100 Bioanalyzer for visualizing the amplified fragments. Use of the PCR combined with the Bioanalyzer offered significant benefits over traditional agarose gel electrophoresis and staining methods.
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