A survey was conducted during 2013 in different regions of Tamil Nadu, India and a total of 51 poultry feed samples were collected from poultry farms and poultry feed dealers. The presence of Aspergillus flavus in the feeds was detected by a polymerase chain reaction (PCR) assay using specific primers based on the O-methyltransferase gene (omt-A) that is involved in the aflatoxin B1 biosynthesis. The contamination levels of Aflatoxin (AFB1, AFB2, AFG1 and AFG2) were determined using high-performance liquid chromatography (HPLC). The results indicated that 98% of the tested samples were contaminated with AFB1 and the levels ranged from 0 to 160.7 ppb and the levels of AFB1 in 29% of the samples exceeded 20 ppb. Aflatoxin B2 contamination was observed in more than 82 % of the samples tested and the levels of

AFB2 in 17 % of the samples exceeded 20 ppb. The levels of AFB2 ranged from 0 to 95.5 ppb. AFG1 and AFG2 were not detected in any of these samples. In the PCR analysis using omt-A primers, A. flavus specific 300-bp PCR product was amplified from DNA extracted from most of the AFB1 contaminated feeds. However, some of the feed samples even though contain AFB1 failed to amplify the 300-bp product in PCR.