Abstract
In the present study, a novel non radioactive digoxigenen labelled PCR-DNA probe was developed targeting nor-1 gene to assess the contamination of aflatoxigenic Aspergillus species in food grain samples from Southern parts of India. The sensitivity of developed PCR-DNA probe was determined to be 10 pg of genomic DNA and 1 pg of purified PCR product. The specificity of the DNA probe was validated by testing against an array of Aspergillus, Fusarium and Penicillium spp. A total of 89 Aspergillus isolates were recovered from 152 grain samples of maize, paddy, and groundnut. Among them, maize had the highest (90%) incidence of toxigenic Aspergillus species. When developed PCR-DNA probe was evaluated onto pure cultures of toxigenic and nontoxigenic Aspergillus species, 60 samples were positive for the nor-1 gene probe. DNA probe results unequivocally matched with the HPLC analysis. In conclusion, the novel PCR-DNA probe developed in this study may find its application in rapid detection of aflatoxin-producing Aspergillus isolates from contaminated cereal grains.
Highlights
Aflatoxins are common mycotoxins in agriculture and food commodities causing serious health hazards to humans and animals leading to great economic losses
Amplification was not observed when these Polymerase Chain Reaction (PCR) assays were employed on other Aspergillus species viz., A. oryzae, A. fumigatus, A. ochraceous and A. carbonarious
In the present study a non-radiactive nor-1 gene DNA probe was designed for specific detection and differentiation of aflatoxigenic and non-toxigenic Aspergillus species
Summary
Aflatoxins are common mycotoxins in agriculture and food commodities causing serious health hazards to humans and animals leading to great economic losses. Molecular techniques have been widely applied for distinguishing aflatoxigenic and non-aflatoxigenic strains of Aspergillus by targeting one or several genes involved in the aflatoxin biosynthetic pathway with the ability/inability to produce aflatoxins. Polymerase Chain Reaction (PCR) methods have been developed for detection of aflatoxigenic fungi. In this respect, several primers have been designed for several genes in the biosynthetic pathways of aflatoxins; for example, afl, nor, omt, ord, tub, ver [5,6,7,8,9]. A novel nonradioactive digoxigenen labelled PCR-DNA probe was developed targeting nor-1 gene to assess the contamination of aflatoxigenic Aspergillus species in contaminated food grain samples. To know the incidence of toxigenic Aspergillus species and aflatoxin concentrations, a sur-
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