Multiple-locus Variable-number Tandem Repeat Analysis for Molecular Typing of Aspergillus Fumigatus

Overview

Journal BMC Microbiol

Publisher Biomed Central

Specialty Microbiology

Date 2010 Dec 15

PMID 21143842

Citations 9

Authors

Simon Thierry

Dongying Wang

Pascal Arne

Manjula Deville

Barbara De Bruin

Adelaide Nieguitsila

Christine Pourcel

Karine Laroucau

Rene Chermette

Weiyi Huang

Francoise Botterel

Jacques Guillot

Affiliations

Soon will be listed here.

Abstract

Background: Multiple-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.

Results: We 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.

Conclusions: MLVA 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.

Citing Articles

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Hadrich I, Khemakhem N, Ilahi A, Trabelsi H, Sellami H, Makni F J Fungi (Basel). 2023; 9(2).

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Aspergillus flavus genetic structure at a turkey farm.

Cherif G, Hadrich I, Harrabi M, Kallel A, Fakhfekh N, Messaoud M Vet Med Sci. 2022; 9(1):234-241.

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Clinical and Microbiological Characterization of Invasive Pulmonary Aspergillosis Caused by in China.

Yu S, Guo L, Xiao M, Zhou M, Yuan Y, Wang Y Front Microbiol. 2020; 11:1672.

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Monitoring of clinical strains and environmental fungal aerocontamination to prevent invasive aspergillosis infections in hospital during large deconstruction work: a protocol study.

Loeffert S, Melloul E, Dananche C, Henaff L, Benet T, Cassier P BMJ Open. 2017; 7(11):e018109.

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

Wang D, Hadj-Henni L, Thierry S, Arne P, Chermette R, Botterel F PLoS One. 2012; 7(9):e44204.

PMID: 23028503 PMC: 3444452. DOI: 10.1371/journal.pone.0044204.

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