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Journal of Clinical Microbiology, July 2009, p. 2079-2083, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.00551-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Aspergillus Section Fumigati Typing by PCR-Restriction Fragment Polymorphism{triangledown}

Janet F. Staab,1 S. Arunmozhi Balajee,2 and Kieren A. Marr1*

Johns Hopkins University School of Medicine, Baltimore, Maryland,1 Mycotic Diseases Branch, Centers for Disease Control and Prevention, Atlanta, Georgia2

Received 18 March 2009/ Returned for modification 14 April 2009/ Accepted 21 April 2009

Recent studies have shown that there are multiple clinically important members of the Aspergillus section Fumigati that are difficult to distinguish on the basis of morphological features (e.g., Aspergillus fumigatus, A. lentulus, and Neosartorya udagawae). Identification of these organisms may be clinically important, as some species vary in their susceptibilities to antifungal agents. In a prior study, we utilized multilocus sequence typing to describe A. lentulus as a species distinct from A. fumigatus. The sequence data show that the gene encoding β-tubulin, benA, has high interspecies variability at intronic regions but is conserved among isolates of the same species. These data were used to develop a PCR-restriction fragment length polymorphism (PCR-RFLP) method that rapidly and accurately distinguishes A. fumigatus, A. lentulus, and N. udagawae, three major species within the section Fumigati that have previously been implicated in disease. Digestion of the benA amplicon with BccI generated unique banding patterns; the results were validated by screening a collection of clinical strains and by in silico analysis of the benA sequences of Aspergillus spp. deposited in the GenBank database. PCR-RFLP of benA is a simple method for the identification of clinically important, similar morphotypes of Aspergillus spp. within the section Fumigati.

* Corresponding author. Mailing address: Johns Hopkins University School of Medicine, 720 Rutland Ave., Ross-1064, Baltimore, MD 21205. Phone: (443) 614-9141. Fax: (443) 614-8518. E-mail: kmarr4{at}jhmi.edu

{triangledown} Published ahead of print on 29 April 2009.

Journal of Clinical Microbiology, July 2009, p. 2079-2083, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.00551-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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