Identification of fungemia agents using the polymerase chain reaction and restriction fragment length polymorphism analysis

Prompt and specific identification of fungemia agents is important in order to define clinical treatment. However, in most cases conventional culture identification can be considered to be time-consuming and not without errors. The aim of the present study was to identify the following fungemia agents: Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata, Cryptococcus neoformans, Cryptococcus gattii, and Histoplasma capsulatum using the polymerase chain reaction and restriction fragment length polymorphism analysis (PCR/RFLP). More specifically: a) to evaluate 3 different amplification regions, b) to investigate 3 different restriction enzymes, and c) to use the best PCR/RFLP procedure to indentify 60 fungemia agents from a culture collection. All 3 pairs of primers (ITS1/ITS4, NL4/ITS5 and Primer1/Primer2) were able to amplify DNA from the reference strains. However, the size of these PCR products did not permit the identification of all the species studied. Three restriction enzymes were used to digest the PCR products: HaeIII, Ddel and Bfal. Among the combinations of pairs of primers and restriction enzymes, only one (primer pair NL4/ITS5 and restriction enzyme Ddel) produced a specific RFLP pattern for each microorganism studied. Sixty cultures of fungemia agents (selected from the culture collection of Fundação de Medicina Tropical do Amazonas - FMTAM) were correctly identified by PCR/RFLP using the prime pair NL4/ITS5 and Ddel. We conclude that the method proved to be both simple and reproducible, and may offer potential advantages over phenotyping methods.

Hanseniase no Estado do Amazonas: historico e desativacao do leprosario. / Hanseniasis in the State of Amazonas: historical aspects and disactivation of the leprosarium

Sao estudados os principais aspectos da evolucao da hanseniase no Estado do Amazonas, desde os primeiros casos registrados, sua provavel procedencia, a criacao dos primeiros isolamentos e a desativacao do leprosario em 1979. Ate dezembro de 1979 estavam registrados 10.800 doentes; desse total, 4.142 eram da forma virchowiana e/ou dimorfa, 4.523 da forma tuberculoide e 2.135 do tipo indeterminado. Os doentes da antiga colonia estao sob controle no posto de saude do atual Bairro do Aleixo ou no asilo, tambem area da antiga colonia