Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens

Brazilian Journal of Infectious Diseases

Cano, Luz¹  Gaviria, Marcela¹  Muñoz-Cadavid, Cesar¹  Rivera, Vanessa¹  Naranjo, Tonny¹ 

[1] Corporación para Investigaciones Biológicas, Medellín, Colombia

关键词: Cryptococcus neoformans/Cryptococcus gattiicomplex;    Nested PCR;    Cryptococcosis;    Molecular diagnosis;   

DOI  :  10.1016/j.bjid.2015.07.006

来源: Contexto

【 摘 要 】

The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity. In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2 regions of rDNA in samples from a cohort of patients diagnosed with cryptococcosis. The results showed that in our hands, this Cryptococcus nested PCR assay has 100% specificity and 100% sensitivity and was able to detect until 2 femtograms of Cryptococcus DNA.

【 授权许可】

Unknown