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 Age I, Bsm FI and Hpa II. 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.