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(Reference retrieved automatically from SciELO through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Visible DNA microarray and loop-mediated isothermal amplification (LAMP) for the identification of Cryptococcus species recovered from culture medium and cerebrospinal fluid of patients with meningitis

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Author(s):
P. Stivanelli [1] ; C.A. Tararam [2] ; P. Trabasso [3] ; L.O. Levy [4] ; M.S.C. Melhem ; A.Z. Schreiber [6] ; M.L. Moretti
Total Authors: 7
Affiliation:
[1] Departamento de Medicina Interna, Faculdade de Ciências Médicas, Universidade Estadual de Campinas - Brasil
[2] Departamento de Medicina Interna, Faculdade de Ciências Médicas, Universidade Estadual de Campinas - Brasil
[3] Departamento de Medicina Interna, Faculdade de Ciências Médicas, Universidade Estadual de Campinas - Brasil
[4] Departamento de Medicina Interna, Faculdade de Ciências Médicas, Universidade Estadual de Campinas - Brasil
[6] Departamento de Patologia Clínica, Faculdade de Ciências Médicas, Universidade Estadual de Campinas - Brasil
Total Affiliations: 7
Document type: Journal article
Source: Brazilian Journal of Medical and Biological Research; v. 53, n. 11 2020-10-09.
Abstract

Cryptococcal meningitis affects normal hosts and immunocompromised patients exhibiting high mortality rates. The objective of this study was to design two molecular assays, visible microarray platforms and loop-mediated isothermal amplification (LAMP), to identify Cryptococcus spp. and the species neoformans and gattii from the cerebral spinal fluid (CSF). To identify Cryptococcus and the two species, we designed two microarrays DNA platforms based on the internal transcribed spacer (ITS) region and CAP59 gene and LAMP assays specific for Cryptococcus species. The assays were tested using CSF from patients with cryptococcal meningitis. CSF from patients with cryptococcal meningitis was cultured in Sabouraud culture medium, and the Cryptococcus spp. grown in the culture medium were also tested for LAMP and microarray platforms. The results were compared to DNA sequencing of the same genetic regions. A total of 133 CSF samples were studied. Eleven CSFs were positive for Cryptococcus (9 C. neoformans and 2 C. gattii), 15 were positive for bacteria, and 107 were negative. The CAP59 platform correctly identified 73% of the CSF samples, while the ITS platform identified 45.5%. CAP59 platform correctly identified 100% of the Cryptococcus isolates, and ITS platform identified 70%. The two sets of LAMP primers correctly identified 100% of the Cryptococcus isolates. However, for CSF samples, the amplification occurred only in 55.5% of C. neoformans. The methodologies were reliable in the identification of Cryptococcus species, mainly for isolates from culture medium, and they might be applied as adjunctive tests to identify Cryptococcus species. (AU)

FAPESP's process: 16/12414-2 - Evaluation of the performance of molecular techniques as DNA microarray and LAMP for diagnosis and accompaniment in patients with cryptococcal meningitis
Grantee:Pamella Stivanelli
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 15/25035-7 - Development and standardization of DNA microarray and Loop mediated isothermal amplification (LAMP) for identification of pathogenic fungi in clinical samples
Grantee:Maria Luiza Moretti
Support Opportunities: Regular Research Grants