Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/7495
Title: Accuracy of real-time PCR, Gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis
Authors: Wu, Henry M.
Cordeiro, Soraia Machado
Harcourt, Brian H
Carvalho, Maria da Gloria S
Azevedo, Jailton
Oliveira, Tainara Queiroz
Leite, Mariela C
Salgado, Kátia M.
Reis, Mitermayer Galvão dos
Plikaytis, Brian D.
Clark, Thomas A
Mayer, Leonard W
Ko, Albert Icksang
Martin, Stacey W
Reis, Joice Neves
Affilliation: National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
Federal University of Bahia. Salvador, BA, Brasil
Federal University of Bahia. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Weill Medical College of Cornell University. New York, NY, USA
National Center for Immunization and Respiratory Diseases CDC. Division of Bacterial Diseases. Atlanta, GA, USA
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. Salvador, BA, Brasil
Abstract: BACKGROUND: Although cerebrospinal fluid (CSF) culture is the diagnostic reference standard for bacterial meningitis, its sensitivity is limited, particularly when antibiotics were previously administered. CSF Gram staining and real-time PCR are theoretically less affected by antibiotics; however, it is difficult to evaluate these tests with an imperfect reference standard. METHODS AND FINDINGS: CSF from patients with suspected meningitis from Salvador, Brazil were tested with culture, Gram stain, and real-time PCR using S. pneumoniae, N. meningitidis, and H. influenzae specific primers and probes. An antibiotic detection disk bioassay was used to test for the presence of antibiotic activity in CSF. The diagnostic accuracy of tests were evaluated using multiple methods, including direct evaluation of Gram stain and real-time PCR against CSF culture, evaluation of real-time PCR against a composite reference standard, and latent class analysis modeling to evaluate all three tests simultaneously. RESULTS: Among 451 CSF specimens, 80 (17.7%) had culture isolation of one of the three pathogens (40 S. pneumoniae, 36 N. meningitidis, and 4 H. influenzae), and 113 (25.1%) were real-time PCR positive (51 S. pneumoniae, 57 N. meningitidis, and 5 H. influenzae). Compared to culture, real-time PCR sensitivity and specificity were 95.0% and 90.0%, respectively. In a latent class analysis model, the sensitivity and specificity estimates were: culture, 81.3% and 99.7%; Gram stain, 98.2% and 98.7%; and real-time PCR, 95.7% and 94.3%, respectively. Gram stain and real-time PCR sensitivity did not change significantly when there was antibiotic activity in the CSF. CONCLUSION: Real-time PCR and Gram stain were highly accurate in diagnosing meningitis caused by S. pneumoniae, N. meningitidis, and H. influenzae, though there were few cases of H. influenzae. Furthermore, real-time PCR and Gram staining were less affected by antibiotic presence and might be useful when antibiotics were previously administered. Gram staining, which is inexpensive and commonly available, should be encouraged in all clinical settings.
Keywords: Bacterial meningitis
Diagnostic test evaluation
Real-time PCR
Streptococcus pneumoniae
Neisseria meningitidis
Haemophilus influenzae
DeCS: Haemophilus influenzae/genética
Meningites Bacterianas/diagnóstico
Neisseria meningitidis/genética
Streptococcus pneumoniae/genética
Adolescente
Adulto
Idoso
Líquido Cefalorraquidiano/microbiologia
Criança
Pré-Escolar
Violeta de Genciana
Haemophilus influenzae/isolamento & purificação
Humanos
Lactente
Recém-Nascido
Meia-Idade
Neisseria meningitidis/isolamento & purificação
Fenazinas
Reação em Cadeia da Polimerase em Tempo Real
Sensibilidade e Especificidade
Streptococcus pneumoniae/isolamento & purificação
Adulto Jovem
Issue Date: 2013
Publisher: BioMed Central
Citation: WU, H. M. et al. Accuracy of real-time PCR, Gram stain and culture for Streptococcus pneumoniae, Neisseria meningitidis and Haemophilus influenzae meningitis diagnosis. BMC Infectious Disease, v. 13, p. 26, 2013.
DOI: 10.1186/1471-2334-13-26.
ISSN: 1471-2334
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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