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Braz. j. infect. dis ; 19(1): 62-67, Jan-Feb/2015. tab, graf
Article in English | LILACS | ID: lil-741236

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 cere- brospinal 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 cryptococcal 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. gattii species complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis. .


Subject(s)
Humans , Cryptococcus gattii/genetics , Cryptococcus neoformans/genetics , DNA, Fungal/cerebrospinal fluid , Meningitis, Cryptococcal/diagnosis , AIDS-Related Opportunistic Infections/cerebrospinal fluid , AIDS-Related Opportunistic Infections/diagnosis , AIDS-Related Opportunistic Infections/microbiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , DNA Primers/genetics , Genotype , Meningitis, Cryptococcal/cerebrospinal fluid , Meningitis, Cryptococcal/microbiology , Polymerase Chain Reaction , Sensitivity and Specificity
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