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Evaluation of the performance of molecular techniques as DNA microarray and lamp for diagnosis and accompaniment in patients with cryptococcal meningitis

Grant number: 16/12414-2
Support type:Scholarships in Brazil - Master
Effective date (Start): September 01, 2016
Effective date (End): August 31, 2018
Field of knowledge:Health Sciences - Medicine
Principal Investigator:Maria Luiza Moretti
Grantee:Pamella Stivanelli
Home Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Cryptococcal meningitis is an infection caused by Cryptococcus yeasts that primarily affects patients infected with HIV, immunocompromised hosts and organ transplant recipientes. Cryptococcus neoformans and Cryptococcus gattii are species that cause meningitis in humans, being the meningitis caused by C. gatti more serious than by C. neoformans, which have been proposed different treatment regimens for each species. Therefore, the main of this study is to evaluate the performance of molecular techniques of DNA microarray and loop mediated isothermal amplification (LAMP) to differentiate C. gattii and C. neoformans directly from clinical samples of cerebrospinal fluid (CSF) of patients with cryptococcal meningitis. Patients at the Clinical Hospital of the University of Campinas, São Paulo, will be included in the study when diagnosed with cryptococcal meningitis by direct microscopy or culture of clinical samples of CSF, performed by the Laboratory of Microbiology. CSF samples with negative cultures for any microorganisms or positive cultures for bacteria with will be used as negative control of reactions. The fungal DNA will be obtained directly from CSF samples and submited to identification by DNA microarray and LAMP. The results will be compared with the identification obtained by DNA sequencing and by conventional microbiology. Cryptococcus spp. isolates in culture will also be tested for sensitivity to antifungal agentes, besides clinical data of patients will be included in the project. For these analyzes, it is intended to establish molecular techniques that can be used to differentiate the diagnosis of meningitis caused by C. gattii or C. neoformans directly from clinical samples, consequently the study of epidemiology of the disease.

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