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

Detalhes bibliográficos
Autor(a) principal: Martins,Marilena dos Anjos
Data de Publicação: 2015
Outros Autores: Brighente,Kate Bastos Santos, Matos,Terezinha Aparecida de, Vidal,Jose Ernesto, Hipólito,Daise Damaris Carnietto de, Pereira-Chioccola,Vera Lucia
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Infectious Diseases
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000100062
Resumo: 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. gattiispecies 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. neoformansand another for C. gattii).Results:CN4-CN5 primer set was positive in all Cryptococcusstandard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattiiwas observed in the clinical samples of either group. The 695 bp products of C. neoformanswere 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. gattiispecies complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.
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spelling Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complexCryptococcal meningitisAcquired immunodeficiency syndromePolymerase chain reactionCerebrospinal fluidAim: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. gattiispecies 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. neoformansand another for C. gattii).Results:CN4-CN5 primer set was positive in all Cryptococcusstandard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattiiwas observed in the clinical samples of either group. The 695 bp products of C. neoformanswere 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. gattiispecies complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.Brazilian Society of Infectious Diseases2015-02-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000100062Brazilian Journal of Infectious Diseases v.19 n.1 2015reponame:Brazilian Journal of Infectious Diseasesinstname:Brazilian Society of Infectious Diseases (BSID)instacron:BSID10.1016/j.bjid.2014.09.004info:eu-repo/semantics/openAccessMartins,Marilena dos AnjosBrighente,Kate Bastos SantosMatos,Terezinha Aparecida deVidal,Jose ErnestoHipólito,Daise Damaris Carnietto dePereira-Chioccola,Vera Luciaeng2015-10-26T00:00:00Zoai:scielo:S1413-86702015000100062Revistahttps://www.bjid.org.br/https://old.scielo.br/oai/scielo-oai.phpbjid@bjid.org.br||lgoldani@ufrgs.br1678-43911413-8670opendoar:2015-10-26T00:00Brazilian Journal of Infectious Diseases - Brazilian Society of Infectious Diseases (BSID)false
dc.title.none.fl_str_mv Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
title Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
spellingShingle Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
Martins,Marilena dos Anjos
Cryptococcal meningitis
Acquired immunodeficiency syndrome
Polymerase chain reaction
Cerebrospinal fluid
title_short Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
title_full Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
title_fullStr Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
title_full_unstemmed Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
title_sort Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex
author Martins,Marilena dos Anjos
author_facet Martins,Marilena dos Anjos
Brighente,Kate Bastos Santos
Matos,Terezinha Aparecida de
Vidal,Jose Ernesto
Hipólito,Daise Damaris Carnietto de
Pereira-Chioccola,Vera Lucia
author_role author
author2 Brighente,Kate Bastos Santos
Matos,Terezinha Aparecida de
Vidal,Jose Ernesto
Hipólito,Daise Damaris Carnietto de
Pereira-Chioccola,Vera Lucia
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Martins,Marilena dos Anjos
Brighente,Kate Bastos Santos
Matos,Terezinha Aparecida de
Vidal,Jose Ernesto
Hipólito,Daise Damaris Carnietto de
Pereira-Chioccola,Vera Lucia
dc.subject.por.fl_str_mv Cryptococcal meningitis
Acquired immunodeficiency syndrome
Polymerase chain reaction
Cerebrospinal fluid
topic Cryptococcal meningitis
Acquired immunodeficiency syndrome
Polymerase chain reaction
Cerebrospinal fluid
description 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. gattiispecies 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. neoformansand another for C. gattii).Results:CN4-CN5 primer set was positive in all Cryptococcusstandard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattiiwas observed in the clinical samples of either group. The 695 bp products of C. neoformanswere 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. gattiispecies complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.
publishDate 2015
dc.date.none.fl_str_mv 2015-02-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000100062
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702015000100062
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1016/j.bjid.2014.09.004
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Brazilian Society of Infectious Diseases
publisher.none.fl_str_mv Brazilian Society of Infectious Diseases
dc.source.none.fl_str_mv Brazilian Journal of Infectious Diseases v.19 n.1 2015
reponame:Brazilian Journal of Infectious Diseases
instname:Brazilian Society of Infectious Diseases (BSID)
instacron:BSID
instname_str Brazilian Society of Infectious Diseases (BSID)
instacron_str BSID
institution BSID
reponame_str Brazilian Journal of Infectious Diseases
collection Brazilian Journal of Infectious Diseases
repository.name.fl_str_mv Brazilian Journal of Infectious Diseases - Brazilian Society of Infectious Diseases (BSID)
repository.mail.fl_str_mv bjid@bjid.org.br||lgoldani@ufrgs.br
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