Development and standardization of DNA microarray and Loop mediated isothermal amplification (LAMP) for identification of pathogenic fungi in clinical samples

Abstract

The identification of the etiologic agent is the key for the early and appropriate use of antifungal therapy. Conventional identification of fungi depends on the combination of microscopic analyzes and growth of the microorganism in culture; these techniques have low sensitivity and delays the identification of pathogen due to the time required for growth. Molecular methods are promising tools offering a rapid and precise identification. The main objectives of this project are: (i) to standardize probes of a DNA microarray platform for the identification of the following fungi: Candida albicans, Candida dubliniensis, Cryptococcus neoformans, Cryptococcus gattii, Fusarium sp., Fusarium solani species complex and Histoplasma capsulatum using isolates grown in culture and from clinical specimens, such as blood culture, whole blood, cerebrospinal fluid and peritoneal liquid; (ii) to develop and standardize a loop mediated isothermal amplification (LAMP) test for the identification of Cryptococcus neoformans and Cryptococcus gattii from cerebral spinal fluid of patients with cryptococcal meningitis. Therefore, the objective of this study is to standardize molecular methodologies for rapid detection and reliable identification of fungal species that cause infections in humans. These tests could be implemented in clinical practice, mainly for direct identification of fungal pathogens from clinical samples. The results of this project can be deposit as patent for future commercialization and application in clinical practice. (AU)