Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.

Aspergillus flavus is second only to A. fumigatus in causing invasive aspergillosis and it is the major agent responsible for fungal sinusitis, keratitis and endophthalmitis in many countries in the Middle East, Africa and Southeast Asia. Despite the growing challenge due to A. flavus, data on the m...

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Autores principales:	Dong Ying Wang, Leila Hadj-Henni, Simon Thierry, Pascal Arné, René Chermette, Françoise Botterel, Inès Hadrich, Fattouma Makni, Ali Ayadi, Stéphane Ranque, Wei Yi Huang, Jacques Guillot
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fc1a9b9867c04cde884d5dceee954c0c
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spelling	oai:doaj.org-article:fc1a9b9867c04cde884d5dceee954c0c2021-11-18T07:05:25ZSimple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.1932-620310.1371/journal.pone.0044204https://doaj.org/article/fc1a9b9867c04cde884d5dceee954c0c2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23028503/?tool=EBIhttps://doaj.org/toc/1932-6203Aspergillus flavus is second only to A. fumigatus in causing invasive aspergillosis and it is the major agent responsible for fungal sinusitis, keratitis and endophthalmitis in many countries in the Middle East, Africa and Southeast Asia. Despite the growing challenge due to A. flavus, data on the molecular epidemiology of this fungus remain scarce. The objective of the present study was to develop a new typing method based on the detection of VNTR (Variable number tandem repeat) markers. Eight VNTR markers located on 6 different chromosomes (1, 2, 3, 5, 7 and 8) of A. flavus were selected, combined by pairs for multiplex amplifications and tested on 30 unrelated isolates and six reference strains. The Simpson index for individual markers ranged from 0.398 to 0.818. A combined loci index calculated with all the markers yielded an index of 0.998. The MLVA (Multiple Locus VNTR Analysis) technique proved to be specific and reproducible. In a second time, a total of 55 isolates from Chinese avian farms and from a Tunisian hospital have been evaluated. One major cluster of genotypes could be defined by using the graphing algorithm termed Minimum Spanning Tree. This cluster comprised most of the isolates collected in an avian farm in southern China. The MLVA technique should be considered as an excellent and cost-effective typing method that could be used in many laboratories without the need for sophisticated equipment.Dong Ying WangLeila Hadj-HenniSimon ThierryPascal ArnéRené ChermetteFrançoise BotterelInès HadrichFattouma MakniAli AyadiStéphane RanqueWei Yi HuangJacques GuillotPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 9, p e44204 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dong Ying Wang Leila Hadj-Henni Simon Thierry Pascal Arné René Chermette Françoise Botterel Inès Hadrich Fattouma Makni Ali Ayadi Stéphane Ranque Wei Yi Huang Jacques Guillot Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
description	Aspergillus flavus is second only to A. fumigatus in causing invasive aspergillosis and it is the major agent responsible for fungal sinusitis, keratitis and endophthalmitis in many countries in the Middle East, Africa and Southeast Asia. Despite the growing challenge due to A. flavus, data on the molecular epidemiology of this fungus remain scarce. The objective of the present study was to develop a new typing method based on the detection of VNTR (Variable number tandem repeat) markers. Eight VNTR markers located on 6 different chromosomes (1, 2, 3, 5, 7 and 8) of A. flavus were selected, combined by pairs for multiplex amplifications and tested on 30 unrelated isolates and six reference strains. The Simpson index for individual markers ranged from 0.398 to 0.818. A combined loci index calculated with all the markers yielded an index of 0.998. The MLVA (Multiple Locus VNTR Analysis) technique proved to be specific and reproducible. In a second time, a total of 55 isolates from Chinese avian farms and from a Tunisian hospital have been evaluated. One major cluster of genotypes could be defined by using the graphing algorithm termed Minimum Spanning Tree. This cluster comprised most of the isolates collected in an avian farm in southern China. The MLVA technique should be considered as an excellent and cost-effective typing method that could be used in many laboratories without the need for sophisticated equipment.
format	article
author	Dong Ying Wang Leila Hadj-Henni Simon Thierry Pascal Arné René Chermette Françoise Botterel Inès Hadrich Fattouma Makni Ali Ayadi Stéphane Ranque Wei Yi Huang Jacques Guillot
author_facet	Dong Ying Wang Leila Hadj-Henni Simon Thierry Pascal Arné René Chermette Françoise Botterel Inès Hadrich Fattouma Makni Ali Ayadi Stéphane Ranque Wei Yi Huang Jacques Guillot
author_sort	Dong Ying Wang
title	Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
title_short	Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
title_full	Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
title_fullStr	Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
title_full_unstemmed	Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.
title_sort	simple and highly discriminatory vntr-based multiplex pcr for tracing sources of aspergillus flavus isolates.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/fc1a9b9867c04cde884d5dceee954c0c
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Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.

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