A Novel Non Radioactive PCR-DNA Probe for the Detection of Aflatoxin Producing Aspergillus Species from Major Food Crops Grown in India

S. R. Priyanka, M. Venkata Ramana, K. Balakrishna, H. S. Murali, H. V. Batra

Division of Microbiology, Defense Food Research Laboratory, Siddarthanagar, India.

DOI: 10.4236/aim.2012.24075                

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.

Keywords

AFB1-Aflatoxin B1; HPLC-High Performance Liquid Chromatography; NOR-1; Norsolorinic Acid Synthase; PCR-Polymerase Chain Reaction

Conflicts of Interest

The authors declare no conflicts of interest.

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