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Journal of Clinical Microbiology, May 2007, p. 1455-1462, Vol. 45, No. 5
0095-1137/07/$08.00+0     doi:10.1128/JCM.00243-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Genotyping of Candida orthopsilosis Clinical Isolates by Amplification Fragment Length Polymorphism Reveals Genetic Diversity among Independent Isolates and Strain Maintenance within Patients{triangledown}

Arianna Tavanti,1*,{dagger} Lambert A. M. Hensgens,1,{dagger} Emilia Ghelardi,2 Mario Campa,2 and Sonia Senesi1

Dipartimento di Biologia,1 Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università di Pisa, Pisa, Italy2

Received 31 January 2007/ Returned for modification 12 February 2007/ Accepted 15 February 2007

Candida parapsilosis former groups II and III have recently been established as independent species named C. orthopsilosis and C. metapsilosis, respectively. In this report, 400 isolates (290 patients) previously classified as C. parapsilosis by conventional laboratory tests were screened by BanI digestion profile analysis of the secondary alcohol dehydrogenase gene fragment and by amplification fragment length polymorphism (AFLP). Thirty-three strains collected from 13 patients were identified as C. orthopsilosis, thus giving the first retrospective evidence that C. orthopsilosis was responsible for 4.5% of the infections/colonization attributed to C. parapsilosis. AFLP was proven to unambiguously identify C. orthopsilosis at the species level and efficiently delineate intraspecific genetic relatedness. A high percentage of polymorphic AFLP bands was observed for independent isolates collected from each patient. Statistical analysis of the pairwise genetic distances and bootstrapping revealed that clonal reproduction and recombination both contribute to C. orthopsilosis genetic population structure. AFLP patterns of sequential isolates obtained from two patients demonstrated that a successful strain colonization within the same patient occurred, as revealed by strain maintenance in various body sites. No association between AFLP markers and drug resistance was observed, and none of the clinical C. orthopsilosis isolates were found to produce biofilm in vitro.

* Corresponding author. Mailing address: Dipartimento di Biologia, Via San Zeno 37, 56127 Pisa, Italy. Phone: 39 050 2213697. Fax: 39 050 2213711. E-mail: atavanti{at}biologia.unipi.it

{triangledown} Published ahead of print on 28 February 2007.

{dagger} These authors contributed equally to the study.

Journal of Clinical Microbiology, May 2007, p. 1455-1462, Vol. 45, No. 5
0095-1137/07/$08.00+0     doi:10.1128/JCM.00243-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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