A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India — Oak Academic Publishing
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A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing <i>Aspergillus</i> Species from Major Food Crops Grown in India
Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
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Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
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Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
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Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
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Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
1 Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
2 Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
3 Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
4 Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
5 Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India
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.
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