Differentiation of Candida species obtained from nosocomial candidemia using RAPD-PCR technique

Abstract

Thirteen strains of the genus Candida were isolated from catheter, urine and surgical wounds from individual patients of the Santa Casa de Misericórdia, Belo Horizonte, MG, Brazil. Ten strains were characterized as Candida albicans, two as Candida glabrata, and one as Candida parapsilosis. Isolates were evaluated for molecular relatedness by random amplified polymorphic DNA technique using 15 primers. The analysis of the genomic DNA obtained revealed a low intraspecific polymorphism and did not allow for the differentiation between strains of the same species obtained from distinct clinical sources (catheter, urine and surgical wounds). The RAPD profiles generated were able to differentiate among the species of Candida albicans, Candida parapsilosis and Candida glabrata strains isolated in this study. Key-words: Candida spp. Random amplified polymorphic DNA. Polymorphism. Nosocomial infection. RESUMO Treze amostras de leveduras do gênero Candida foram isoladas de catéter, urina e feridas cirúrgicas de pacientes da Santa Casa de Misericórida de Belo Horizonte, MG, Brasil. Dez amostras foram identificadas como Candida albicans, duas comoCandida glabrata e uma como Candida parapsilosis. Os isolados foram avaliados quanto ao perfil molecular pela técnica de amplificação aleatória de DNA polimórfico utilizando 15 iniciadores. A análise do DNA genômico obtido revelou um baixo polimorfismo intraespecífico e não permitiu a diferenciação entre amostras da mesma espécie obtidas a partir de diferentes espécimes clínicos (catéter, urina e feridas cirúrgicas). Os perfis de RAPD obtidos foram capazes de diferenciar entre as espécies Candida albicans, Candida parapsilosis e Candida glabrata isoladas neste estudo. Palavras-chaves: Candida spp. Amplificação aleatória de DNA polimórfico. Polimorfismo. Infecção nosocomial.

Molecular diagnosis of vaginal candidiasis by polymerase chain reaction (PCR) and random amplification polymorphism DNA (RAPD-PCR) in Babylon Province, Iraq

“…RAPD-PCR was performed with total volume of 50 μl consisting of 10 μl of 50x master mix (Promiga, USA ) with 37 μl molecular-grade water, 0.5 μl template DNA and 1.5 μl (50 pmole) for each random primers RP1-4: TAGGATCAGA and SOY: AGGTCACTGA (Valerio et al, 2006). Amplified PCR conditions were as follows: 95°C for 4 min, 38 cycle 94°C for 1 min, 36°C for 1.5 min, 72°C for 1.5 min.…”

Oral Candida spp. colonization in human immunodeficiency virus-infected individuals

“…It is easy and rapid but presents reproducibility problems inter and intra laboratories (107). There are reports about its application for several Candida species (86,116). Recently, two new techniques arouse: amplified fragment length polymorphism (AFLP) (3) and multilocus sequence typing (MLST) (12,32,115).…”

Identification and differentiation of Candida species from pediatric patients by random amplified polymorphic DNA

“…Apart from corroborating previous studies that reported failure of microbiological techniques to identify Candida species 1 22 , the present study has successfully identified Candida isolates. It was able to discriminate between species through the use of a single primer, while other studies have reported success in Candida species typing through the use of a set of primers 24 . We conclude that RAPD with the primer OP-G10 could be performed after culture isolation, in order to confirm microbiological species identification, thus providing faster and more reliable Candida species typing.…”