Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis

© 2018 Society of Chemical Industry. BACKGROUND: Aspergillus is a diverse genus of fungi with high economic and social impact. Various species that belong to section Nigri (black aspergilli) are common agents of grape spoilage and potent producers of ochratoxin A (OTA), a mycotoxin associated with v...

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Main Authors: Aliki Xanthopoulou, Ioannis Ganopoulos, Paschalitsa Tryfinopoulou, Efstathios Z. Panagou, Maslin Osanthanunkul, Panagiotis Madesis, Dimosthenis Kizis
Format: Journal
Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/58175
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-581752018-09-05T04:37:50Z Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis Aliki Xanthopoulou Ioannis Ganopoulos Paschalitsa Tryfinopoulou Efstathios Z. Panagou Maslin Osanthanunkul Panagiotis Madesis Dimosthenis Kizis Agricultural and Biological Sciences Biochemistry, Genetics and Molecular Biology Nursing © 2018 Society of Chemical Industry. BACKGROUND: Aspergillus is a diverse genus of fungi with high economic and social impact. Various species that belong to section Nigri (black aspergilli) are common agents of grape spoilage and potent producers of ochratoxin A (OTA), a mycotoxin associated with various nephrotoxic and immunotoxic effects in humans. Black aspergilli are difficult to classify following only phenotypic criteria; thus chemotaxonomic and molecular methods are employed in parallel with phenotypic ones for species characterization. These approaches, though accurate and replicable, require more than one individual step and are to a certain extent laborious when a rapid identification of these species is required. RESULTS: The aim of this study was to develop a high-resolution melting polymerase chain reaction (HRM-PCR) assay as a rapid method for identification of Aspergillus spp. section Nigri isolates and their detection in grape samples. Melt curve analysis of amplicons originating from the internal transcribed spacer 2 (ITS2) ribosomal region generated species-specific HRM curve profiles, enabling the accurate differentiation of the analyzed genotypes. Furthermore, the assay was able to identify A. carbonarius, A. tubingensis, A. niger, A. ibericus and A. japonicus in grape samples artificially inoculated with conidia of these fungi. CONCLUSION: To our knowledge this is the first report on the development of an HRM-PCR assay for the identification of black Aspergillus species in grape samples. 2018-09-05T04:20:41Z 2018-09-05T04:20:41Z 2018-01-01 Journal 10970010 00225142 2-s2.0-85050453197 10.1002/jsfa.9189 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85050453197&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/58175
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Agricultural and Biological Sciences
Biochemistry, Genetics and Molecular Biology
Nursing
spellingShingle Agricultural and Biological Sciences
Biochemistry, Genetics and Molecular Biology
Nursing
Aliki Xanthopoulou
Ioannis Ganopoulos
Paschalitsa Tryfinopoulou
Efstathios Z. Panagou
Maslin Osanthanunkul
Panagiotis Madesis
Dimosthenis Kizis
Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
description © 2018 Society of Chemical Industry. BACKGROUND: Aspergillus is a diverse genus of fungi with high economic and social impact. Various species that belong to section Nigri (black aspergilli) are common agents of grape spoilage and potent producers of ochratoxin A (OTA), a mycotoxin associated with various nephrotoxic and immunotoxic effects in humans. Black aspergilli are difficult to classify following only phenotypic criteria; thus chemotaxonomic and molecular methods are employed in parallel with phenotypic ones for species characterization. These approaches, though accurate and replicable, require more than one individual step and are to a certain extent laborious when a rapid identification of these species is required. RESULTS: The aim of this study was to develop a high-resolution melting polymerase chain reaction (HRM-PCR) assay as a rapid method for identification of Aspergillus spp. section Nigri isolates and their detection in grape samples. Melt curve analysis of amplicons originating from the internal transcribed spacer 2 (ITS2) ribosomal region generated species-specific HRM curve profiles, enabling the accurate differentiation of the analyzed genotypes. Furthermore, the assay was able to identify A. carbonarius, A. tubingensis, A. niger, A. ibericus and A. japonicus in grape samples artificially inoculated with conidia of these fungi. CONCLUSION: To our knowledge this is the first report on the development of an HRM-PCR assay for the identification of black Aspergillus species in grape samples.
format Journal
author Aliki Xanthopoulou
Ioannis Ganopoulos
Paschalitsa Tryfinopoulou
Efstathios Z. Panagou
Maslin Osanthanunkul
Panagiotis Madesis
Dimosthenis Kizis
author_facet Aliki Xanthopoulou
Ioannis Ganopoulos
Paschalitsa Tryfinopoulou
Efstathios Z. Panagou
Maslin Osanthanunkul
Panagiotis Madesis
Dimosthenis Kizis
author_sort Aliki Xanthopoulou
title Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
title_short Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
title_full Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
title_fullStr Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
title_full_unstemmed Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis
title_sort rapid and accurate identification of black aspergilli from grapes using high-resolution melting (hrm) analysis
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85050453197&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/58175
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