Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus

Human mpox is caused by the Monkeypox virus, a microorganism closely related to the Variola virus, both belonging to the Orthopoxvirus genus. Mpox had been considered a rare disease until a global outbreak occurred in 2022. People infected with the virus present similar symptoms to patients sufferin...

Descripción completa

Detalles Bibliográficos
Autores: Martínez-Murcia, Antonio, Navarro, Aaron, García-Sirera, A., Pérez, Laura, Bru, G.
Tipo de recurso: artículo
Fecha de publicación:2023
País:España
Institución:Universidad Miguel Hernández de Elche
Repositorio:REDIUMH. Depósito Digital de la UMH
OAI Identifier:oai:dspace.umh.es:11000/35094
Acceso en línea:https://hdl.handle.net/11000/35094
Access Level:acceso abierto
Palabra clave:Monkeypox virus
qPCR
Detection
Validation
UNE/EN ISO/IEC 17025:2005
id ES_fe49347984e1e45909ffde9320ec856d
oai_identifier_str oai:dspace.umh.es:11000/35094
network_acronym_str ES
network_name_str España
repository_id_str
spelling Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virusMartínez-Murcia, AntonioNavarro, AaronGarcía-Sirera, A.Pérez, LauraBru, G.Monkeypox virusqPCRDetectionValidationUNE/EN ISO/IEC 17025:2005Human mpox is caused by the Monkeypox virus, a microorganism closely related to the Variola virus, both belonging to the Orthopoxvirus genus. Mpox had been considered a rare disease until a global outbreak occurred in 2022. People infected with the virus present similar symptoms to patients suffering smallpox and other rash illnesses, hindering diagnosis. The WHO indicated that no commercial PCR or serology kits are currently widely available. In the present study, the MPXV MONODOSE dtec-qPCR kit was validated following guidelines of the UNE/EN ISO/IEC 17025:2005. The parameters evaluated for the acceptance of the assay were in silico and in vitro specificity, quantitative phase analysis, reliability, and sensitivity. The assay passed validation criteria and yielded an efficiency of 95.8%, high repeatability, reproducibility, and a Limit of Detection and Quantification of at least 10 copies. Results from the validation of the MPXV dtec-qPCR kit were satisfactory. The use of the MONODOSE format (dehydrated single PCR-tubes, ready to use) provided considerable advantages allowing the detection of the Monkeypox virus to be accurately achieved. This detection kit may be considered a reliable, fast, simple, and universally available optionMDPIDepartamentos de la UMH::Producción Vegetal y Microbiología202520252023info:eu-repo/semantics/articleapplication/pdf11application/pdfhttps://hdl.handle.net/11000/35094reponame:REDIUMH. Depósito Digital de la UMHinstname:Universidad Miguel Hernández de ElcheIngléshttps://doi.org/10.3390/diagnostics13091560info:eu-repo/semantics/openAccessAttribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/oai:dspace.umh.es:11000/350942026-05-27T13:36:21Z
dc.title.none.fl_str_mv Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
title Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
spellingShingle Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
Martínez-Murcia, Antonio
Monkeypox virus
qPCR
Detection
Validation
UNE/EN ISO/IEC 17025:2005
title_short Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
title_full Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
title_fullStr Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
title_full_unstemmed Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
title_sort Internal Validation of a Real-Time qPCR Kit following the UNE/EN ISO/IEC 17025:2005 for Detection of the Re-Emerging Monkeypox virus
dc.creator.none.fl_str_mv Martínez-Murcia, Antonio
Navarro, Aaron
García-Sirera, A.
Pérez, Laura
Bru, G.
author Martínez-Murcia, Antonio
author_facet Martínez-Murcia, Antonio
Navarro, Aaron
García-Sirera, A.
Pérez, Laura
Bru, G.
author_role author
author2 Navarro, Aaron
García-Sirera, A.
Pérez, Laura
Bru, G.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Departamentos de la UMH::Producción Vegetal y Microbiología
dc.subject.none.fl_str_mv Monkeypox virus
qPCR
Detection
Validation
UNE/EN ISO/IEC 17025:2005
topic Monkeypox virus
qPCR
Detection
Validation
UNE/EN ISO/IEC 17025:2005
description Human mpox is caused by the Monkeypox virus, a microorganism closely related to the Variola virus, both belonging to the Orthopoxvirus genus. Mpox had been considered a rare disease until a global outbreak occurred in 2022. People infected with the virus present similar symptoms to patients suffering smallpox and other rash illnesses, hindering diagnosis. The WHO indicated that no commercial PCR or serology kits are currently widely available. In the present study, the MPXV MONODOSE dtec-qPCR kit was validated following guidelines of the UNE/EN ISO/IEC 17025:2005. The parameters evaluated for the acceptance of the assay were in silico and in vitro specificity, quantitative phase analysis, reliability, and sensitivity. The assay passed validation criteria and yielded an efficiency of 95.8%, high repeatability, reproducibility, and a Limit of Detection and Quantification of at least 10 copies. Results from the validation of the MPXV dtec-qPCR kit were satisfactory. The use of the MONODOSE format (dehydrated single PCR-tubes, ready to use) provided considerable advantages allowing the detection of the Monkeypox virus to be accurately achieved. This detection kit may be considered a reliable, fast, simple, and universally available option
publishDate 2023
dc.date.none.fl_str_mv 2023
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/11000/35094
url https://hdl.handle.net/11000/35094
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.3390/diagnostics13091560
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.format.none.fl_str_mv application/pdf
11
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:REDIUMH. Depósito Digital de la UMH
instname:Universidad Miguel Hernández de Elche
instname_str Universidad Miguel Hernández de Elche
reponame_str REDIUMH. Depósito Digital de la UMH
collection REDIUMH. Depósito Digital de la UMH
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869425671524057088
score 15,811543

Similares en LA Referencia