Journal article
A real-time reverse-transcriptase PCR technique for detection and quantification of viable Alternaria spp. in foodstuffs
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain
A real-time reverse-transcriptase PCR (RT-PCR) technique was developed for the rapid and specific detection and enumeration of viable Alternaria spp. in foodstuffs. The method uses Alernaria-specific primers and probe targeting the internal transcribed spacer regions ITS1 and ITS2 of the rRNA gene. The detection limit of the real-time RT-PCR assay to detect viable Alternaria spp. in food samples was 1 CFU/g.
The estimated Alternaria counts obtained by real-time RT-PCR showed a good correlation (R2 = 0.9881, P < 0.01) in the range of 1–105 CFU/mL with the Alternaria counts obtained by culture methods. The applicability of the real-time RT-PCR protocol was assessed through analysis of 110 commercial food samples, including 60 fresh fruit and vegetable samples and 50 processed foodstuffs.
The assay developed provides a useful tool for early detection of low concentrations of viable Alternaria spp. in naturally contaminated food samples, and could be applied as a quality and biosecurity marker of raw materials and final products in the fruits and vegetables processing industries.
Language: | English |
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Year: | 2012 |
Pages: | 286-294 |
ISSN: | 18737129 and 09567135 |
Types: | Journal article |
DOI: | 10.1016/j.foodcont.2012.05.017 |