Keywords
diagnosis
real time PCR
Reagent Kits
diagnostic
How to Cite
Abstract
Bacterial meningitis (BM) remains a worldwide problem and a severe threat to public health. The purpose of this study was to evaluate the performance of three commercial kits (ALLPLEX™ Meningitis-B Assay/Seegene Inc., South Korea; VIASURE™ H.influenzae, N.meningitidis & S.pneumoniae Real Time PCR Detection Kit/CerTest Biotec, Spain; XGEN™ Multi MB/Mobius Life Science, Brazil) for the simultaneous detection of Neisseria meningitidis (Nm), Streptococcus pneumoniae (Spn) and Haemophillus influenzae(Hi). The performance of three commercial kits was compared with “in house” molecular assay routinely used at the Instituto Adolfo Lutz/SP/Brazil (IAL). Both commercial and “in house” assays are based on multiplex real time PCR assay (mqPCR). The mqPCR standardized by IAL (mqPCR/IAL) was introduced in BM diagnostic routine since 2010, and it was considered as reference test in this study. A total of 100 clinical samples (cerebrospinal fluid= 62; serum= 38) were tested using the three commercial mqPCR kits in comparison with mqPCR/IAL assay, being 93 positives for Nm, Spn or Hi, and 7 negatives. For DNA-CSF samples, the results from ALLPLEX™, VIASURE™, and XGEN™ kits showed high agreement with mqPCR/IAL assay, presenting more than 93% of concordance and Kappa index between 0,61 and 0,80. For DNA-serum samples, only VIASURE™ kit was compared with mqPCR/IAL assay, presenting 100% of concordance similarity and Kappa index = 1. All the three kits proved to be competent for qualitative detection of Nm, Spn and Hi and they are able to be used in the routine diagnostics of BM.
References
Brouwer MC, Tunkel AR, van de Beek D. Epidemiology, diagnosis, and antimicrobial treatment of acute bacterial meningitis. Clin microbiol rev. 2010; 23(3): 467-92. https://doi.org/10.1128/cmr.00070-09
Bottomley MJ, Serruto D, Sáfadi MAP, Klugman KP. Future challenges in the elimination of bacterial meningitis. Vaccine. 2012; 30S: B78-B86. https://doi.org/10.1016/j.vaccine.2011.12.099
L. Liphaus B, Cobo Zanella R, Paula S. Lemos A, C. G. Almeida S, Chimara E, M. Blanco R, T. Higa F, Gisele Gonçalves M, M. Salgado M, O. Fukasawa L, Cristina Pereira dos Santos F, R.M.P. Carvalhanas T. Meningites Bacterianas: Diagnóstico e Caracterização Laboratorial dos Agentes Etiológicos. Bepa [Internet]. 30º de novembro de 2021 [citado 2º de março de 2023];18(215):69-86. Disponível em: https://periodicos.saude.sp.gov.br/BEPA182/article/view/37236
Marques Pinto Carvalhanas TR, de Cunto Brandileone MC, Cobo Zanella R. Meningites Bacterianas. Bepa [Internet]. 31º de maio de 2005 [citado 2º de março de 2023];2(17):15-26. Disponível em: https://periodicos.saude.sp.gov.br/BEPA182/article/view/38902
Obaro S. Updating the diagnosis of bacterial meningitis. Lancet infect dis. 2019; 19: 1160-61. https://doi: 10.1016/S1473-3099(19)30549-3
Sacchi CT, Fukasawa LO, Gonçalves MG, Salgado MM, Shutt KA, Carvalhanas TR, et al. Incorporation of real-time PCR into routine public health surveillance of culture negative bacterial meningitis in São Paulo, Brazil. PLoS One. 2011; 6(6): e20675. https://doi.org/10.1371/journal.pone.0020675. Epub 2011 Jun22. PMID: 21731621; PMCID: PMC3120771.
Salgado MM, Higa FT, Gonçalves MG, Fukasawa LO, Liphaus BL, Oliveira PL, et al. Nova versão do ensaio da PCR em tempo real para o diagnóstico laboratorial e vigilância epidemiológica das meningites bacterianas. BEPA – Boletim Epidemiológico Paulista. 2012; 9(103): 16-20. Disponível em: https://docs.bvsalud.org/biblioref/ses-sp/2012/ses-28039/ses-28039-4714.pdf
Emery SL, Erdman DD, Bowen MD, Newton BR, Winchell JM, Meyer RF, et al. Real-time reverse transcription-polymerase chain reaction assay for SARS-associated coronavirus. Emerg infect dis. 2004 Feb; 10(2): 311-16. https://doi.org/10.3201/eid1002.030759. PMID: 15030703; PMCID: PMC3322901.
Landis, JR; Koch, GG. The measurement of observer agreement for categorical data. Biometrics. 1977; 33 (1): 159-74. https://doi.org/10.2307%2F2529310
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Copyright (c) 2023 Maria Gisele Gonçalves, Fábio Takenori Higa, Lucila Okuyama Fukasawa , Gabriela Andrade de Carvalho , Bruno Silva Milagres , Maristela Marques Salgado