Multiplex polymerase chain reaction assay for the differential detection of trichothecene- and fumonisin-producing species of Fusarium in cornmeal

The genus Fusarium comprises a diverse group of fungi including several species that produce mycotoxins in food commodities. In this study, a multiplex polymerase chain reaction (PCR) assay was developed for the group-specific detection of fumonisin-producing and trichothecene-producing species of F...

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Bibliographic Details
Published in:Journal of food protection 2002-12, Vol.65 (12), p.1955-1961
Main Authors: BLUHM, B. H, FLAHERTY, J. E, COUSIN, M. A, WOLOSHUK, C. P
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cereal and baking product industries
Consumer Product Safety
DNA Primers
DNA, Fungal - genetics
DNA, Intergenic
Food industries
Food Microbiology
Fumonisins
Fundamental and applied biological sciences. Psychology
Fusarium - isolation & purification
Fusarium - metabolism
Humans
Mycotoxins - biosynthesis
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Species Specificity
Trichothecenes - biosynthesis
Trichothecenes - chemistry
Zea mays - microbiology
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Summary:The genus Fusarium comprises a diverse group of fungi including several species that produce mycotoxins in food commodities. In this study, a multiplex polymerase chain reaction (PCR) assay was developed for the group-specific detection of fumonisin-producing and trichothecene-producing species of Fusarium. Primers for genus-level recognition of Fusarium spp. were designed from the internal transcribed spacer regions (ITS1 and ITS2) of rDNA. Primers for group-specific detection were designed from the TRI6 gene involved in trichothecene biosynthesis and the FUM5 gene involved in fumonisin biosynthesis. Primer specificity was determined by testing for cross-reactivity against purified genomic DNA from 43 fungal species representing 14 genera, including 9 Aspergillus spp., 9 Fusarium spp., and 10 Penicillium spp. With purified genomic DNA as a template, genus-specific recognition was observed at 10 pg per reaction; group-specific recognition occurred at 100 pg of template per reaction for the trichothecene producer Fusarium graminearum and at 1 ng of template per reaction for the fumonisin producer Fusarium verticillioides. For the application of the PCR assay, a protocol was developed to isolate fungal DNA from cornmeal. The detection of F. graminearum and its differentiation from F. verticillioides were accomplished prior to visible fungal growth at
ISSN:0362-028X
1944-9097
DOI:10.4315/0362-028x-65.12.1955