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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.

brazilian journal of medical and biological research

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