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Journal of Clinical Microbiology, August 2005, p. 4112-4120, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4112-4120.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of a Novel Panel of Nine Short Tandem Repeats for Exact and High-Resolution Fingerprinting of Aspergillus fumigatus Isolates

Hanneke A. de Valk,1 Jacques F. G. M. Meis,1 Ilse M. Curfs,1 Konrad Muehlethaler,2 Johan W. Mouton,1 and Corné H. W. Klaassen1*

Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands,1 Institute for Infectious Diseases, University of Bern, Bern, Switzerland2

Received 14 January 2005/ Returned for modification 28 March 2005/ Accepted 22 April 2005

Here we describe a new panel of short tandem repeats (STRs) for a novel exact typing assay that can be used to discriminate between Aspergillus fumigatus isolates. A total of nine STR markers were selected from available genomic A. fumigatus sequences and were divided into three multicolor multiplex PCRs. Each multiplex reaction amplified three di-, tri-, or tetranucleotide repeats, respectively. All nine STR markers were used to analyze 100 presumably unrelated A. fumigatus isolates. For each marker, between 11 and 37 alleles were found in this population. One isolate proved to be a mixture of at least two different isolates. With the remaining 99 isolates, 96 different fingerprinting profiles were obtained. The Simpson's diversity index for the individual markers ranged from 0.77 to 0.97. The diversity index for the multiplex combination of di-, tri-, and tetranucleotide repeats ranged from 0.9784 to 0.9968. The combination of all nine markers yielded a Simpson's diversity index of 0.9994, indicative of the high discriminatory power of these new loci. In theory, this panel of markers is able to discriminate between no less than 27 x 109 different genotypes. The multicolor multiplex approach allows large numbers of markers to be tested in a short period of time. The exact nature of the assay combines high reproducibility with the easy exchange of results and makes it a very suitable tool for large-scale epidemiological studies.

* Corresponding author. Mailing address: Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands. Phone: 31-24-3657514. Fax: 31-24-3657516. E-mail: c.klaassen{at}cwz.nl.

Journal of Clinical Microbiology, August 2005, p. 4112-4120, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4112-4120.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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