Hyd5 gene-based detection of the major gushing-inducing Fusarium spp. in a loop-mediated isothermal amplification (LAMP) assay

International Journal of Food Microbiology
Carla DenschlagLudwig Niessen

Abstract

Fusarium graminearum and the closely related F. culmorum were found to be associated with over foaming of bottled beer (gushing) when contaminated brewing malt is used. The presence of highly surface active hydrophobins produced by these fungi upon growth on wheat or barley in the field or during malting may affect bubble formation and stability in gushing beers and other carbonated beverages. Aiming for a method for the rapid and user friendly analysis of unmalted and malted cereals during quality control in the brewing industry, a loop-mediated isothermal amplification (LAMP) assay for the detection of Fusarium spp. capable of producing the gushing inducing hydrophobin Hyd5p was set up. A set of primers was designed towards a 221 bp region within the hyd5 gene of F. culmorum. The LAMP product was verified by sequencing a 150 bp portion. Testing specificity with purified DNA from 99 different fungal species as well as barley and wheat showed that DNA synthesis only occurred during LAMP when DNA of the closely related species F. graminearum, F. culmorum, F. cerealis and F. lunulosporum were used as template. In-tube indirect detection of DNA amplification was applied using manganese-quenched calcein as fluorescence indicator fo...Continue Reading

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Citations

Feb 26, 2013·Journal of Agricultural and Food Chemistry·Felix FockeMarkus Fischer
Sep 21, 2013·Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology·Ya-Lan ZengXue-Jun Ma
Jan 3, 2013·Molecular Plant Pathology·Barbara SchermQuirico Migheli
Nov 14, 2012·International Journal for Parasitology·Xuelian LiJing Yuan
Sep 3, 2014·Biotechnology, Biotechnological Equipment·Mohammad A KhiyamiKamel A Abd-Elsalam

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