title: RAPD analysis with the primer L15996 of Brazilian clinical and environmental Cryptococcus neoformans isolates.
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.
authors.affiliation: Infectious Diseases Unit, Triangulo Mineiro Federal University, Avenida Frei Paulino, 30, Uberaba, Minas Gerais, CEP 38025-180, Brazil. leonardoeuripedes@gmail.com
authors.forename: Leonardo
authors.initials: L
authors.lastname: Andrade-Silva
authors.forename: Kennio
authors.initials: K
authors.lastname: Ferreira-Paim
authors.forename: Delio Jose
authors.initials: DJ
authors.lastname: Mora
authors.forename: Paulo Roberto
authors.initials: PR
authors.lastname: da Silva
authors.forename: Anderson Assunção
authors.initials: AA
authors.lastname: Andrade
authors.forename: Eliane
authors.initials: E
authors.lastname: Lages-Silva
authors.forename: André Luiz
authors.initials: AL
authors.lastname: Pedrosa
authors.forename: Mario León
authors.initials: ML
authors.lastname: Silva-Vergara
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001938:Brazil; D016000:Cluster Analysis; D003453:Cryptococcosis / Q000382:microbiology*; D003455:Cryptococcus neoformans / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004783:Environmental Microbiology; D005656:Fungal Proteins / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D058889:Molecular Typing; D016533:Mycological Typing Techniques; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D037521:Virulence Factors / Q000235:genetics
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D037521:Virulence Factors