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Extraction of DNA to detection of aflatoxigenic aspergillus species in foodstuffs using PCR method
Authors: Šnévajsová Petra | Brožková Iveta | Vytřasová Jarmila
Year: 2007
Type of publication: článek v odborném periodiku
Name of source: Scientific Papers of the University of Pardubice, Series A, Faculty of Chemical Technology
Publisher name: Univerzita Pardubice
Place: Pardubice
Page from-to: 87?97
Titles:
Language Name Abstract Keywords
cze Extrakce DNA z aflatoxinogenních druhů aspergilů vyskytujících se v potravinách pro PCR Extrakce DNA z aflatoxinogenních druhů aspergilů vyskytujících se v potravinách pro následnou polymerázovou řetězovou reakci. Aspergillus flavus; ektrakce DNA; PCR
eng Extraction of DNA to detection of aflatoxigenic aspergillus species in foodstuffs using PCR method The extraction of DNA according to Cenis was used with different modifications of disrupting the cell wall. Fungi mycelium was disrupted by ultrasonic bath, Eppendorf homogenizer and liquid nitrogen. Method was optimized using pure cultures A. parasiticus var. globosus CCM F ? 550 and A. flavus CCM F ? 108. After extraction, acquired DNA was amplified by Polymerase Chain Reaction (PCR). PCR was used to amplify ver-1 gene and apa-2 gene as target fragments. The specific PCR product codes the biosynthetic way of aflatoxin B1. PCR product was detected by electrophoresis. Records were interpreted. Acquired results demonstrate, that success of method depends especially on disrupting cell wall of fungi. 91 samples of food (first of all tea, spices and medical herbs) were examined. 27 samples were positive for the presence of the aflatoxinogenic fungi on the AFPA medium. Two samples contained two species fungi of genus Aspergillus and one sample contained three species fungi of genus Aspergillus. Finally we determined 31 kinds of the potentially aflatoxinogenic fungi on the AFPA medium. The PCR were positive for 27 kinds of the aflatoxinogenic fungi Aspergillus flavus; DNA extraction; PCR

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