Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases

Isolation and determination of s.pneumoniae by culture and serological methods can be time consuming or indeterminate. Molecular diagnosis by real-time PCR is independent of the growth of the pathogen causing meningitis, and is not diminished with non-viable organisms. The aim of this study was to e...

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Autores principales:	Chaiboub Soumaya, Charof Réda, Qasmaoui Aicha, Hamamouchi Jamila, Berny El Hassan
Formato:	article
Lenguaje:	EN FR
Publicado:	EDP Sciences 2021
Materias:	Environmental sciences GE1-350
Acceso en línea:	https://doaj.org/article/0c58b05f32f34deca297679adbe0bb11
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spelling	oai:doaj.org-article:0c58b05f32f34deca297679adbe0bb112021-11-12T11:44:08ZPerformance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases2267-124210.1051/e3sconf/202131901063https://doaj.org/article/0c58b05f32f34deca297679adbe0bb112021-01-01T00:00:00Zhttps://www.e3s-conferences.org/articles/e3sconf/pdf/2021/95/e3sconf_vigisan_01063.pdfhttps://doaj.org/toc/2267-1242Isolation and determination of s.pneumoniae by culture and serological methods can be time consuming or indeterminate. Molecular diagnosis by real-time PCR is independent of the growth of the pathogen causing meningitis, and is not diminished with non-viable organisms. The aim of this study was to evaluate the performance criteria of pre-PCR-TR DNA extraction step and PCR-TR step by targeting two genes encoding s.pneumoniae. In this study we evaluated the inter-sample contamination of the pre-PCR-TR step, the intermediate fidelity and the repeatability of the DNA essay. PCR-TR verification was performed by two genes targeting s. pneumoniae the Lyt A and SP 2038 gene; sensitivity, specificity and LLD were determined. Contamination rate had a value of less than 0%, which is in agreement with an absence of inter-sample contamination; the repeatability and intermediate fidelity have a cv˂7%. The evaluation of the sensitivity and specificity of the RT-PCR assays targeted 100% the Lyt A gene and the SP 2038 gene. The standard curve generated detected less than 10copies for the Lyt A gene and less than 100copies for the SP 2038 gene. This study showed that the pre-PCR and PCR-TR assays met the performance criteria targeted in this study.Chaiboub SoumayaCharof RédaQasmaoui AichaHamamouchi JamilaBerny El HassanEDP SciencesarticleEnvironmental sciencesGE1-350ENFRE3S Web of Conferences, Vol 319, p 01063 (2021)
institution	DOAJ
collection	DOAJ
language	EN FR
topic	Environmental sciences GE1-350
spellingShingle	Environmental sciences GE1-350 Chaiboub Soumaya Charof Réda Qasmaoui Aicha Hamamouchi Jamila Berny El Hassan Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
description	Isolation and determination of s.pneumoniae by culture and serological methods can be time consuming or indeterminate. Molecular diagnosis by real-time PCR is independent of the growth of the pathogen causing meningitis, and is not diminished with non-viable organisms. The aim of this study was to evaluate the performance criteria of pre-PCR-TR DNA extraction step and PCR-TR step by targeting two genes encoding s.pneumoniae. In this study we evaluated the inter-sample contamination of the pre-PCR-TR step, the intermediate fidelity and the repeatability of the DNA essay. PCR-TR verification was performed by two genes targeting s. pneumoniae the Lyt A and SP 2038 gene; sensitivity, specificity and LLD were determined. Contamination rate had a value of less than 0%, which is in agreement with an absence of inter-sample contamination; the repeatability and intermediate fidelity have a cv˂7%. The evaluation of the sensitivity and specificity of the RT-PCR assays targeted 100% the Lyt A gene and the SP 2038 gene. The standard curve generated detected less than 10copies for the Lyt A gene and less than 100copies for the SP 2038 gene. This study showed that the pre-PCR and PCR-TR assays met the performance criteria targeted in this study.
format	article
author	Chaiboub Soumaya Charof Réda Qasmaoui Aicha Hamamouchi Jamila Berny El Hassan
author_facet	Chaiboub Soumaya Charof Réda Qasmaoui Aicha Hamamouchi Jamila Berny El Hassan
author_sort	Chaiboub Soumaya
title	Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
title_short	Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
title_full	Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
title_fullStr	Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
title_full_unstemmed	Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases
title_sort	performance verification of pre-pcr and real-time pcr step in the molecular diagnosis of pneumococcal meningitis cases
publisher	EDP Sciences
publishDate	2021
url	https://doaj.org/article/0c58b05f32f34deca297679adbe0bb11
work_keys_str_mv	AT chaiboubsoumaya performanceverificationofprepcrandrealtimepcrstepinthemoleculardiagnosisofpneumococcalmeningitiscases AT charofreda performanceverificationofprepcrandrealtimepcrstepinthemoleculardiagnosisofpneumococcalmeningitiscases AT qasmaouiaicha performanceverificationofprepcrandrealtimepcrstepinthemoleculardiagnosisofpneumococcalmeningitiscases AT hamamouchijamila performanceverificationofprepcrandrealtimepcrstepinthemoleculardiagnosisofpneumococcalmeningitiscases AT bernyelhassan performanceverificationofprepcrandrealtimepcrstepinthemoleculardiagnosisofpneumococcalmeningitiscases
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Performance verification of pre-PCR and real-time PCR step in the molecular diagnosis of pneumococcal meningitis cases

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