Enzyme-Linked Immunosorbent Assay (ELISA) for Aflatoxin B1 Estimation in Groundnuts

Anjaiah, V and Mehan, V K and Jayanthi, S and Reddy, D V R and McDonald, D (1989) Enzyme-Linked Immunosorbent Assay (ELISA) for Aflatoxin B1 Estimation in Groundnuts. In: International Workshop on Aflatoxin Contamination of Groundnut, 6-9 Oct 1987, Patancheru, Andhra Pradesh, India.

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The commercially available hapten, afla B1-oxime-bovine serum albumin, was used to produce an antiserum in rabbits. The same hapten was coupled with alkaline phosphatase (hapten-BSA-ALP) and used in the competitive direct enzyme-linked immunosorbent assay (ELISA) for the detection of aflatoxin B1. Aflatoxin B1 was extracted in methanol from naturally contaminated or 'spiked' groundnut seed samples. Wells of a polystyrene microtitre plate were coated with the antiserum, the plates were washed in PBS-Tween, aflatoxin B1 standards or groundnut sample extracts, and hapten-BSA-ALP conjugate were added and the plates incubated. The plates were again washed, and the amount of conjugate bound to the antibody was determined after addition of the substrate, p-nitro-phenylphosphate. The hapten-BSA-ALP conjugate has advantages in stability, simplicity of preparation, and high specificity, over the conventional toxin-enzyme conjugate in direct competitive ELISA. The assay method is more rapid and less expensive than the physico-chemical methods of aflatoxin analysis and it can detect levels of aflatoxin B1 as low as 50 picograms.

Item Type: Conference or Workshop Item (Paper)
Subjects: Mandate crops > Groundnut
Depositing User: Library ICRISAT
Date Deposited: 04 Dec 2011 12:00
Last Modified: 04 Dec 2011 12:00
URI: http://oar.icrisat.org/id/eprint/4481
Acknowledgement: UNSPECIFIED
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