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Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples

Streptococcus pneumoniae causes invasive diseases of significant public health concern, such as meningitis. The culture of cerebrospinal fluid (CSF) samples, the standard technique for meningitis diagnoses, is not always positive. Consequently, meaningful information about the etiological agent is l...

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Autores principales: de Souza, Mariana Brena, de Carvalho, Eneas, Cergole-Novella, Maria Cecília, Molinari, Delma Aparecida, Colpas, Daniela Rodrigues, dos Santos Carmo, Andréia Moreira, dos Santos Menezes Gaiotto Daros, Vilma, de Campos, Ivana Barros
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9854411/
https://www.ncbi.nlm.nih.gov/pubmed/36662376
http://dx.doi.org/10.1007/s10096-023-04547-3
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author de Souza, Mariana Brena
de Carvalho, Eneas
Cergole-Novella, Maria Cecília
Molinari, Delma Aparecida
Colpas, Daniela Rodrigues
dos Santos Carmo, Andréia Moreira
dos Santos Menezes Gaiotto Daros, Vilma
de Campos, Ivana Barros
author_facet de Souza, Mariana Brena
de Carvalho, Eneas
Cergole-Novella, Maria Cecília
Molinari, Delma Aparecida
Colpas, Daniela Rodrigues
dos Santos Carmo, Andréia Moreira
dos Santos Menezes Gaiotto Daros, Vilma
de Campos, Ivana Barros
author_sort de Souza, Mariana Brena
collection PubMed
description Streptococcus pneumoniae causes invasive diseases of significant public health concern, such as meningitis. The culture of cerebrospinal fluid (CSF) samples, the standard technique for meningitis diagnoses, is not always positive. Consequently, meaningful information about the etiological agent is lost, which can compromise effective epidemiological surveillance and the improvement of immunization policies. This study aims to standardize a method to genotype pneumococcus in the CSF samples which could mitigate the absence of isolated strains, and also evaluate the prediction of this assay. We applied eight multiplex PCR (mPCR) assays to CSF samples paired with the Quellung reaction applied to the isolated strains. We also compared different master mix kits in the mPCR. Moreover, a retrospective study was conducted with CSF samples considered pneumococcus positive due to the presence of the lytA gene. Results showed that genotyping by the mPCR correlated 100% with the Quellung reaction, and genotyping was dependent on the master mix applied. In the retrospective study (2014–2020), 73.4% were successfully genotyped. The analyses of the receiver operating characteristic curve showed that the cycle threshold (Ct value) around 30 for the lytA gene had a 75% positive chance of successful genotyping, whereas with a Ct value > 35, the chance was 12.5%. Finally, we observed that genotype 19A was prevalent in the period (12%), information unknown until now due to the lack of isolated strains. Therefore, the mPCR of CSF samples can efficiently predict S. pneumoniae serotypes, especially in the absence of isolated strains, which can be a great tool for pneumococcal serotype surveillance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10096-023-04547-3.
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spelling pubmed-98544112023-01-23 Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples de Souza, Mariana Brena de Carvalho, Eneas Cergole-Novella, Maria Cecília Molinari, Delma Aparecida Colpas, Daniela Rodrigues dos Santos Carmo, Andréia Moreira dos Santos Menezes Gaiotto Daros, Vilma de Campos, Ivana Barros Eur J Clin Microbiol Infect Dis Original Article Streptococcus pneumoniae causes invasive diseases of significant public health concern, such as meningitis. The culture of cerebrospinal fluid (CSF) samples, the standard technique for meningitis diagnoses, is not always positive. Consequently, meaningful information about the etiological agent is lost, which can compromise effective epidemiological surveillance and the improvement of immunization policies. This study aims to standardize a method to genotype pneumococcus in the CSF samples which could mitigate the absence of isolated strains, and also evaluate the prediction of this assay. We applied eight multiplex PCR (mPCR) assays to CSF samples paired with the Quellung reaction applied to the isolated strains. We also compared different master mix kits in the mPCR. Moreover, a retrospective study was conducted with CSF samples considered pneumococcus positive due to the presence of the lytA gene. Results showed that genotyping by the mPCR correlated 100% with the Quellung reaction, and genotyping was dependent on the master mix applied. In the retrospective study (2014–2020), 73.4% were successfully genotyped. The analyses of the receiver operating characteristic curve showed that the cycle threshold (Ct value) around 30 for the lytA gene had a 75% positive chance of successful genotyping, whereas with a Ct value > 35, the chance was 12.5%. Finally, we observed that genotype 19A was prevalent in the period (12%), information unknown until now due to the lack of isolated strains. Therefore, the mPCR of CSF samples can efficiently predict S. pneumoniae serotypes, especially in the absence of isolated strains, which can be a great tool for pneumococcal serotype surveillance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10096-023-04547-3. Springer Berlin Heidelberg 2023-01-20 2023 /pmc/articles/PMC9854411/ /pubmed/36662376 http://dx.doi.org/10.1007/s10096-023-04547-3 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Article
de Souza, Mariana Brena
de Carvalho, Eneas
Cergole-Novella, Maria Cecília
Molinari, Delma Aparecida
Colpas, Daniela Rodrigues
dos Santos Carmo, Andréia Moreira
dos Santos Menezes Gaiotto Daros, Vilma
de Campos, Ivana Barros
Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title_full Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title_fullStr Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title_full_unstemmed Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title_short Multiplex PCR to Streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
title_sort multiplex pcr to streptococcus pneumoniae serotype identification directly in cerebrospinal fluid samples
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9854411/
https://www.ncbi.nlm.nih.gov/pubmed/36662376
http://dx.doi.org/10.1007/s10096-023-04547-3
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