Avaliação de kits comerciais baseados em PCR multiplex em tempo real para diagnóstico de meningite bacteriana.

Maria Gisele Gonçalves; Fabio Takenori Higa; Lucila Okuyama Fukasawa; Gabriela Andrade  Carvalho; Bruno Silva Milagres; Maristela Marques Salgado

doi:10.57148/bepa.2023.v.20.39209

Authors

Maria Gisele Gonçalves Instituto Adolfo Lutz https://orcid.org/0000-0002-8966-5927
Fabio Takenori Higa Instituto Adolfo Lutz https://orcid.org/0000-0003-4419-9616
Lucila Okuyama Fukasawa Instituto Adolfo Lutz https://orcid.org/0000-0002-6688-0528
Gabriela Andrade Carvalho Ministério da Saúde https://orcid.org/0009-0006-5888-5587
Bruno Silva Milagres Ministério da Saúde https://orcid.org/0009-0009-8466-806X
Maristela Marques Salgado Instituto Adolfo Lutz https://orcid.org/0000-0002-2085-4965

DOI:

https://doi.org/10.57148/bepa.2023.v.20.39209

Keywords:

bacterial meningitis, diagnosis, real time PCR, Reagent Kits, diagnostic

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.

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