High genetic variability in non-aflatoxigenic A. flavus strains by using Quadruplex PCR-based assay

Department of Microbiological, Genetic and Molecular Sciences, University of Messina, Italy.
International Journal of Food Microbiology (Impact Factor: 3.08). 06/2008; 125(3):341-3. DOI: 10.1016/j.ijfoodmicro.2008.04.020
Source: PubMed


Aflatoxigenic Aspergillus flavus isolates always show, by using a multiplex PCR-system, four DNA fragments specific for aflR, nor-1, ver-1, and omt-A genes. Non-aflatoxigenic A. flavus strains give variable DNA banding pattern lacking one, two, three or four of these genes. Recently, it has been found and reported that some aflatoxin non-producing A. flavus strains show a complete set of genes. Because less is known about the incidence of structural genes aflR, nor-1, ver-1 and omt-A in aflatoxin non-producing strains of A. flavus, we decided to study the frequencies of the aflatoxin structural genes in non-aflatoxigenic A. flavus strains isolated from food and feed commodities. The results can be summarized as following: 36.5% of the examined non-aflatoxigenic A. flavus strains showed DNA fragments that correspond to the complete set of genes (quadruplet pattern) as found in aflatoxigenic A. flavus. Forty three strains (32%) showed three DNA banding patterns grouped in four profiles where nor-1, ver-1 and omt-A was the most frequent profile. Twenty five (18.7%) of non-aflatoxigenic A. flavus strains yielded two DNA banding pattern whereas sixteen (12%) of the strains showed one DNA banding pattern. In one strain, isolated from poultry feed, no DNA bands were found. The nor-1 gene was the most representative between the four aflatoxin structural assayed genes. Lower incidence was found for aflR gene. Our data show a high level of genetic variability among non-aflatoxigenic A. flavus isolates that require greater attention in order to design molecular experiment to distinguish true aflatoxigenic from non-aflatoxigenic A. flavus strains.

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    • "The primers and their sequences (Table 1) which were used in the PCR amplification experiments were reported in previous work [1, 5, 14]. The primers were obtained from Alpha DNA/Canada. "
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