Multiple-locus variable-number tandem repeat analysis for molecular typing of Aspergillus fumigatus

BMC Microbiology

Research Article

Karine Laroucau¹  Simon Thierry²  Dongying Wang³  René Chermette⁴  Jacques Guillot⁴  Barbara De Bruin⁴  Adélaïde Nieguitsila⁴  Pascal Arné⁴  Manjula Deville⁴  Weiyi Huang⁵  Françoise Botterel⁶  Christine Pourcel⁷ 

[1] ANSES, Bacterial Zoonoses Unit, Maisons-Alfort, France;ANSES, UMR BIPAR, Ecopham, Agence Nationale de Sécurité Sanitaire, Maisons-Alfort, France;ANSES, UMR BIPAR, Ecopham, Agence Nationale de Sécurité Sanitaire, Maisons-Alfort, France;Parasitology department, College of Animal Science and Technology, Guangxi University, Nanning, China;ENVA, UMR BIPAR, Ecopham, Ecole nationale vétérinaire d'Alfort, Maisons-Alfort, France;Parasitology department, College of Animal Science and Technology, Guangxi University, Nanning, China;UPE, UMR BIPAR, Ecopham, Faculté de Médecine de Créteil, France;Université Paris Sud 11, CNRS, UMR 8621, Institut de Génétique et Microbiologie, 91405, Orsay, France;

关键词: Amplify Fragment Length Polymorphism;    Aspergillosis;    Minimum Span Tree;    Invasive Aspergillosis;    Poultry Farm;   

DOI  :  10.1186/1471-2180-10-315

 received in 2010-07-07, accepted in 2010-12-08,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundMultiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) is a prominent subtyping method to resolve closely related microbial isolates to provide information for establishing genetic patterns among isolates and to investigate disease outbreaks. The usefulness of MLVA was recently demonstrated for the avian major pathogen Chlamydophila psittaci. In the present study, we developed a similar method for another pathogen of birds: the filamentous fungus Aspergillus fumigatus.ResultsWe selected 10 VNTR markers located on 4 different chromosomes (1, 5, 6 and 8) of A. fumigatus. These markers were tested with 57 unrelated isolates from different hosts or their environment (53 isolates from avian species in France, China or Morocco, 3 isolates from humans collected at CHU Henri Mondor hospital in France and the reference strain CBS 144.89). The Simpson index for individual markers ranged from 0.5771 to 0.8530. A combined loci index calculated with all the markers yielded an index of 0.9994. In a second step, the panel of 10 markers was used in different epidemiological situations and tested on 277 isolates, including 62 isolates from birds in Guangxi province in China, 95 isolates collected in two duck farms in France and 120 environmental isolates from a turkey hatchery in France. A database was created with the results of the present study http://minisatellites.u-psud.fr/MLVAnet/. Three major clusters of isolates were defined by using the graphing algorithm termed Minimum Spanning Tree (MST). The first cluster comprised most of the avian isolates collected in the two duck farms in France, the second cluster comprised most of the avian isolates collected in poultry farms in China and the third one comprised most of the isolates collected in the turkey hatchery in France.ConclusionsMLVA displayed excellent discriminatory power. The method showed a good reproducibility. MST analysis revealed an interesting clustering with a clear separation between isolates according to their geographic origin rather than their respective hosts.

【 授权许可】

Unknown   
© Thierry et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.