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RAPD Analysis with the Primer L15996 of Brazilian Clinical and Environmental Cryptococcus neoformans Isolates

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Abstract

Different methods have been used to perform the molecular characterization of Cryptococcus neoformans. Among them, RAPD analysis is able to separate isolates of the same species and genotypes. This study aimed to evaluate clinical and environmental C. neoformans isolates from Minas Gerais, Brazil by RAPD and correlate the genetic profiles with the ones obtained by URA5-RFLP, virulence factors and antifungal susceptibility patterns. Forty-five environmental (31 from areas surrounding hospital and 14 from captive bird droppings from pet-shops) and 29 clinical C. neoformans isolates were evaluated. Antifungal susceptibility tests (Clinical and Laboratory Standards Institute), URA5-RFLP analysis and the assessment of virulence factors were performed according to their original descriptions. RAPD profiles were obtained using the L15996 primer (5′-CTCCACCATTAGCACCCAAAGC-3′). RAPD analysis generated two to 20 bands for all studied isolates. The isolates presented similarities ranging from 10.8 to 100.0%. Considering a minimum identity score of 50%, four clusters were formed. Cluster I contained 10 pet-shops bird dropping isolates, cluster II contained 22 clinical isolates most of them recovered from cerebrospinal fluid, cluster III contained 14 isolates from hospital surroundings and cluster IV contained 12 environmental isolates most from hospital surroundings. Fourteen isolates were not grouped. The RAPD profiles were clustered according to their source and URA5-RFLP pattern. No correlation between virulence factors or antifungal susceptibility profile with the obtained RAPD profiles was observed.

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Acknowledgments

We thank Miss Angela Azor for her technical assistance, Dr. Claudia Leite Maffei and Dr. Lúcia Kioko Hasimoto e Souza for kindly providing the reference strains of the eight major molecular types of the C. neoformans/C. gattii species complex. Also, we are grateful with Professor Wieland Meyer for the valuable comments and suggestions to improve the manuscript. This work was supported by a Grant Edital Universal APQ-01735/2010 form the Fundação de Amparo à Pesquisa de Minas Gerais FAPEMIG to Mario León Silva-Vergara.

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Authors and Affiliations

  1. Infectious Diseases Unit, Triangulo Mineiro Federal University, Avenida Frei Paulino, 30, Uberaba, Minas Gerais, CEP 38025-180, Brazil

    Leonardo Andrade-Silva, Kennio Ferreira-Paim, Delio Jose Mora, Paulo Roberto da Silva, Anderson Assunção Andrade, Eliane Lages-Silva, André Luiz Pedrosa & Mario León Silva-Vergara

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  1. Leonardo Andrade-Silva
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  2. Kennio Ferreira-Paim
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Correspondence to Mario León Silva-Vergara.

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Andrade-Silva, L., Ferreira-Paim, K., Mora, D.J. et al. RAPD Analysis with the Primer L15996 of Brazilian Clinical and Environmental Cryptococcus neoformans Isolates. Mycopathologia 174, 53–59 (2012). https://doi.org/10.1007/s11046-011-9515-6

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  • DOI: https://doi.org/10.1007/s11046-011-9515-6

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