Rapid immunological assays for detection of aflatoxin B1 in animal feed and its products

Abstract

Aspergillus flavus toxigenic strain was macroscopically and microscopically examined and cultivated on yeast extract sucrose (YES) medium and incubated at 27°C ± 2°C for 15 days. After 15 days, aflatoxin B1 was extracted using chloroform and aflatoxin B1 was determined using high performance liquid chromatography (HPLC) that assured the presence of aflatoxin Bl in a concentration of 22.2 µg / 100 ml YES. As is well known that aflatoxins are toxic fungal metabolites of low molecular weight, and hence are devoid of antigenicity. Those toxins also lack a reactive group for coupling with a macromolecule carrier for antibody production. However, through derivation, a free carboxylic group is introduced to the toxin molecule, consequently permitting the molecule to react covalently with proteins (BSA) to produce the high molecular weight AFB1- BSA conjugate (the immunogen) which will be used in the immunization of the New Zealand rabbits. The present study was planned to produce polyclonal antibodies against aflatoxin B1 as a basic step toward the preparation