Detection and Quantitation of Aflatoxin for the Diagnosis of Aspergillus flavus

G Shrestha1, A Mridha2

Author address: 

1Department of Microbiology, Amrit Campus, Tribhuvan University, Kathmandu, Nepal 2Department of Botany, University of Chittagong, Chittagong, Bangladesh

Abstract: 

Purpose:

Mycotoxins have been known to cause acute kidney failure (orchratoxin), damage of central nervous system (tremorgenic mycotoxin) and damage of the upper respiratory tract. Aflatoxins produced by Aspergillus flavus is a known carcinogenic hepatotoxin. The immunological assays for the detection and quantitation of toxin provide a technique for the diagnosis of disease. Aflatoxin from A. flavus was screened for its detection and quantitation by immunoassay.

Methods:

A. flavus producing mycotoxin were screened on Aflatoxin Producing Medium (APM) as described Donald et al (1981). Extraction of Aflatoxins were carried out (Abarca et al, 1988 and Chu, 1987) and detection of Aflatoxin B1 was done by TLC using pre-coated silica gel plates (Merck) and further confirmed by AOAC (Association of Official Analytical Chemists) method (Horwitz, 1975). Quantitation of Aflatoxin was done by ELISA (Chu, 1987). The data were analyzed by SPSS 16.0.

Results:

The highest amounts of Aflatoxin B1 were reported in synthetic medium, YES (68.56ng/ml and 200ng/ml) with 2% sucrose, pH 5.5 after 14 days of inoculation at 28 0C (p-value 0.05), and in natural medium, groundnuts (121.20ng/g and 500ng/kg) by ELISA and TLC methods respectively.

Conclusion:

Alfatoxins produced by Aspergillus species could be used as a marker for the diagnosis of disease. The spores of A. flavus under the favourable environmental conditions produce sufficient amount of Aflatoxins B1 that was detected and quantified by ELISA and TLC. Detection and quantification of these adducts by immunoassays have been suggested to be an alternative methods to detect human exposure to aflatoxins in blood samples.

Key words: Aflatoxin, Aspergillus flavus, Immunoassays, TLC, ELISA

2012

abstract No: 

14

Full conference title: 

5th Advances Against Aspergillosis
    • AAA 5th (2012)