Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex

Author(s)

Martins, Marilena dos Anjos ^{[1, 2]} ; Santos Brighente, Kate Bastos ^[1] ; de Matos, Terezinha Aparecida ^[3] ; Vidal, Jose Ernesto ^[4] ; Carnietto de Hipolito, Daise Damaris ^[2] ; Pereira-Chioccola, Vera Lucia ^[1]

Affiliation

^[1] Adolfo Lutz Inst, Lab Biol Mol Parasitas & Fungos, BR-01246902 Sao Paulo - Brazil

^[2] Adolfo Lutz Inst, Nucleo Micol, BR-01246902 Sao Paulo - Brazil

^[3] Inst Infectol Emilio Ribas, Lab Liquido Cefalorraquiano, Sao Paulo - Brazil

^[4] Inst Infectol Emilio Ribas, Dept Neurol, Sao Paulo - Brazil

• Document type: Journal article
• Source: Brazilian Journal of Infectious Diseases; v. 19, n. 1, p. 62-67, JAN-FEB 2015.
• Web of Science Citations: 4

Abstract

Aim: This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples. Materials and methods: The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. gattii species complex reference strains and 165 cerebrospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS; and 69 patients with other neurological opportunistic diseases (CRL/AIDS). Two primer sets were tested (CN4-CN5 and the multiplex CNa70S-CNa70A/CNb49S-CNb-49A that amplify a specific product for C. neoformans and another for C. gattii). Results: CN4-CN5 primer set was positive in all Cryptococcus standard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattii was observed in the clinical samples of either group. The 695 bp products of C. neoformans were observed only in 64.6% of the czyptococcal meningitis and AIDS group. This primer set was negative for two standard strains. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets. Conclusions: These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. neoformans/C. gotta species complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis. (C) 2014 Elsevier Editora Ltda. Este e um artigo Open Access sob a licenca de CC BY-NC-ND (AU)

• FAPESP's process: 11/13939-8 - Study of a group of fungal and parasitic infections aiming the improvement of techniques of characterization and immune-molecular diagnosis
• Grantee: Vera Lúcia Pereira Chioccola
• Support type: Regular Research Grants