Development of an ELISA-based method for testing aflatoxigenicity and aflatoxigenic variability among Aspergillus species in culture

Abstract

Aflatoxins contaminate foodstuff posing a severe threat to human health because chronic exposure is linked to liver cancer while acute exposure may cause death. Therefore, it is of interest to reduce the contamination of crops by aflatoxins in the field and post-harvest. Among the current technologies being developed is the deployment of non-aflatoxigenic strains of Aspergillus species to competitively exclude aflatoxigenic conspecifics from crops in the field thereby curtailing aflatoxin production by the former. The success in this endeavor makes the non-aflatoxigenic fungi good candidates for biological control programs. However, the current techniques for segregating non-aflatoxigenic from aflatoxigenic fungi suffer two main drawbacks: they are based on morphological and chemical tests with a combination of visual color changes detected in a culture plate which suffer some degree of inaccuracy. Secondly, the existing methods are incapable of accurately quantifying aflatoxin production by fungi in culture. We developed a culture system for inducing aflatoxin production by Aspergillus using maize kernels as growth substrate followed by quantification using ELISA. The method was compared to the Dichlorvos-Ammonia (DV-AM) method for determining aflatoxigenicity. Our findings encapsulate a method more robust than the currently used DV-AM approach because, for the first time, we are able to assess aflatoxigenicity and aflatoxigenic variability among Aspergillus species earlier classified as non-aflatoxigenic by the DV-AM method. Furthermore, the new method presents an opportunity to attribute the toxin production by actively growing fungal cultures. We believe this method when further developed presents a chance to study and predict fungal behavior prior to field trials for biological control programs. Keywords: ELISA, Aflatoxin, Aspergillus, Aflatoxigenic variability, Aflatoxigenicity, Dichlorvos-Ammonia (DV-AM)