title: PCR-REA as an important tool for the identification of Cryptococcus neoformans and Cryptococcus gattii from human and veterinary sources.
abstract: The extraordinary ability of Cryptococcus species to cause disease has focused the attention of scientists on finding ways to improve their identification methods. In this study, PCR-REA, manual methods (morphological and biochemical characteristics), API 20C and VITEK 2 were used to test identify a total of 30 Cryptococcus spp. from human and veterinary sources. PCR-REA was performed using the capsular region as amplification target followed by restriction with the enzymes AgeI, BsmFI and HpaII. PCR-REA identified the strains as C. neoformans var. grubii (n=19) and C. gattii (n=8). There was no significant difference between the API 20C AUX and VITEK 2 when compared to manual methods for the identification of Cryptococcus spp. However, none of these non-manual methods were able to detect C. gattii samples. PCR-REA showed a greater level of concordance with the manual method, besides being faster and more sensitive than the other methods. Therefore, it is indicated for routine identification of Cryptococcus spp. strains.
authors.affiliation: Department of Pathology and Legal Medicine, School of Medicine, Specialized Medical Mycology Center, Federal University of Ceará, Fortaleza, CE, Brazil. rossanacordeiro@ufc.br
authors.forename: Rossana Aguiar
authors.initials: RA
authors.lastname: Cordeiro
authors.forename: Ana Karoline Freire
authors.initials: AK
authors.lastname: Costa
authors.forename: Raimunda Sâmia Nogueira
authors.initials: RS
authors.lastname: Brilhante
authors.forename: Rita Amanda Chaves
authors.initials: RA
authors.lastname: de Lima
authors.forename: Débora de Souza Collares Maia
authors.initials: Dde S
authors.lastname: Castelo-Branco
authors.forename: Joyce Fonteles
authors.initials: JF
authors.lastname: Ribeiro
authors.forename: André Jalles
authors.initials: AJ
authors.lastname: Monteiro
authors.forename: Francisco Airton Castro
authors.initials: FA
authors.lastname: Rocha
authors.forename: José Júlio Costa
authors.initials: JJ
authors.lastname: Sidrim
authors.forename: Marcos Fábio Gadelha
authors.initials: MF
authors.lastname: Rocha
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods; D056285:Cryptococcus gattii / Q000145:classification / Q000235:genetics*; D003455:Cryptococcus neoformans / Q000145:classification / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D000091402:Prohibitins; D015183:Restriction Mapping / Q000379:methods*
chemical_list: C087781:CAP59 protein, Cryptococcus neoformans; D004271:DNA, Fungal; D005656:Fungal Proteins; C000717331:PHB2 protein, human; D000091402:Prohibitins