Comparison of genotyping methods for Cunninghamella bertholletiae.


Journal

Mycoses
ISSN: 1439-0507
Titre abrégé: Mycoses
Pays: Germany
ID NLM: 8805008

Informations de publication

Date de publication:
Jun 2019
Historique:
received: 25 06 2018
revised: 30 01 2019
accepted: 18 02 2019
pubmed: 3 3 2019
medline: 10 8 2019
entrez: 3 3 2019
Statut: ppublish

Résumé

Invasive fungal infections caused by filamentous fungi of the order Mucorales are serious complications in immunocompromised patients and often associated with fatal outcome. As a member of this order, Cunninghamella bertholletiae is a saprophytic fungus with naturally exhibited high minimum inhibitory concentrations against common antifungal drugs and with the potential for outbreaks in clinical settings. In a proof-of-principle study, we evaluated the performance of microsatellite markers for the discrimination of thirteen C. bertholletiae isolates from various sources in comparison with a repetitive sequence-based PCR (rep-PCR) and random amplification of polymorphic DNA (RAPD). Based on the higher discriminatory power of the microsatellite PCR with five separate primer pairs (Simpson's index of 1 vs 0 [RAPD] and 0 [rep-PCR]), the novel method was applied to eight additional isolates, including four well-characterised isolates from a cluster of infections in a next step. In total, microsatellite PCR identified 21 separate genotypes. A probable epidemiological association of the cluster isolates could be demonstrated by microsatellite genotyping. In conclusion, our findings demonstrate the value of microsatellite PCR in genotyping Cunninghamella bertholletiae and its potential for future applications with other species of the order Mucorales.

Sections du résumé

BACKGROUND BACKGROUND
Invasive fungal infections caused by filamentous fungi of the order Mucorales are serious complications in immunocompromised patients and often associated with fatal outcome. As a member of this order, Cunninghamella bertholletiae is a saprophytic fungus with naturally exhibited high minimum inhibitory concentrations against common antifungal drugs and with the potential for outbreaks in clinical settings.
OBJECTIVES AND METHODS OBJECTIVE
In a proof-of-principle study, we evaluated the performance of microsatellite markers for the discrimination of thirteen C. bertholletiae isolates from various sources in comparison with a repetitive sequence-based PCR (rep-PCR) and random amplification of polymorphic DNA (RAPD). Based on the higher discriminatory power of the microsatellite PCR with five separate primer pairs (Simpson's index of 1 vs 0 [RAPD] and 0 [rep-PCR]), the novel method was applied to eight additional isolates, including four well-characterised isolates from a cluster of infections in a next step.
RESULTS RESULTS
In total, microsatellite PCR identified 21 separate genotypes. A probable epidemiological association of the cluster isolates could be demonstrated by microsatellite genotyping.
CONCLUSION CONCLUSIONS
In conclusion, our findings demonstrate the value of microsatellite PCR in genotyping Cunninghamella bertholletiae and its potential for future applications with other species of the order Mucorales.

Identifiants

pubmed: 30825344
doi: 10.1111/myc.12908
doi:

Types de publication

Comparative Study Evaluation Study Journal Article

Langues

eng

Pagination

519-525

Informations de copyright

© 2019 Blackwell Verlag GmbH.

Auteurs

Hedda Luise Verhasselt (HL)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Julia Radke (J)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Dirk Schmidt (D)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

David Killengray (D)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Ulrike Scharmann (U)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Volker Rickerts (V)

FG 16: Erreger von Pilz-, und Parasiteninfektionen und Mykobakteriosen, Konsiliarlabor für Kryptokokkose und seltene Systemmykosen, Robert Koch-Institut, Berlin, Germany.

Wiebke Hansen (W)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Danila Seidel (D)

Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany.

Iker Falces-Romero (I)

Clinical Microbiology Department, Hospital Universitario La Paz, Madrid, Spain.

Jan Buer (J)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Peter-Michael Rath (PM)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Joerg Steinmann (J)

Institute of Medical Microbiology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.
Institute of Clinical Hygiene, Medical Microbiology and Infectiology, Hospital Nuremberg, Paracelsus Medical University, Nuremberg, Germany.

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Classifications MeSH