Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family

Detalhes bibliográficos
Autor(a) principal: Costa, Cátia
Data de Publicação: 2022
Outros Autores: Ferreira, Guilherme D., Simões, Marco, Silva, Joana L., Campos, Maria J.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.8/8708
Resumo: This work was funded by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within the scope of the project MARE (UIDB/04292/2020 and UIDP/04292/2020) and grant agreement n. POCI-01-0247-FEDER-035234; AlgaValor, Portugal 2020 program.
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spelling Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae familyqPCRVibrio alginolyticusListonella anguillaraVibrio harveyiVibriosisAquacultureThis work was funded by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within the scope of the project MARE (UIDB/04292/2020 and UIDP/04292/2020) and grant agreement n. POCI-01-0247-FEDER-035234; AlgaValor, Portugal 2020 program.Vibriosis, an often-fatal disease induced by pathogenic members of the Vibrionaceae family, causes severe economic losses in aquacultures. To mitigate/avoid vibriosis outbursts, it is vital to detect and quantify these pathogens as early as possible. However, standard microbiological methods are time-consuming and often underestimate cell counts, which calls for the development of valid alternatives. In this study, real-time polymerase chain reaction (qPCR) was employed to detect the pathogenic species Vibrio alginolyticus, Listonella anguillara, and Vibrio harveyi using a new primer pair targeting the groEL gene. In addition, the DNA extraction efficiency of three methods, two commercial kits and the boiling method, was compared. The most efficient method was the DNeasy Blood and Tissue kit, with a detection limit ranging between 154 and 600 CFU mL-1 in the case of V. alginolyticus and L. anguillara, and 48 CFU mL-1 for V. harveyi. Thus, this study presents the development and evaluation of a method for the early quantification of all three species in saline suspensions. However, the results obtained by spiking a microalgae sample with V. harveyi emphasize the importance of adjusting the DNA control’s standard curve to the relevant extraction matrices, as it affects the DNA extraction efficiency and may hamper an accurate quantification with qPCR.MDPIIC-OnlineCosta, CátiaFerreira, Guilherme D.Simões, MarcoSilva, Joana L.Campos, Maria J.2023-07-28T15:12:57Z20222022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.8/8708engCosta, C.; Ferreira, G.D.; Simões, M.; Silva, J.L.; Campos, M.J. Real-Time PCR Protocol for Detection and Quantification of Three Pathogenic Members of the Vibrionaceae Family. Microorganisms 2022, 10, 2060. https://doi.org/ 10.3390/microorganisms101020602076-260710.3390/microorganisms10102060info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-01-17T15:58:00Zoai:iconline.ipleiria.pt:10400.8/8708Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T01:51:22.563936Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
title Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
spellingShingle Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
Costa, Cátia
qPCR
Vibrio alginolyticus
Listonella anguillara
Vibrio harveyi
Vibriosis
Aquaculture
title_short Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
title_full Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
title_fullStr Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
title_full_unstemmed Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
title_sort Real-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae family
author Costa, Cátia
author_facet Costa, Cátia
Ferreira, Guilherme D.
Simões, Marco
Silva, Joana L.
Campos, Maria J.
author_role author
author2 Ferreira, Guilherme D.
Simões, Marco
Silva, Joana L.
Campos, Maria J.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv IC-Online
dc.contributor.author.fl_str_mv Costa, Cátia
Ferreira, Guilherme D.
Simões, Marco
Silva, Joana L.
Campos, Maria J.
dc.subject.por.fl_str_mv qPCR
Vibrio alginolyticus
Listonella anguillara
Vibrio harveyi
Vibriosis
Aquaculture
topic qPCR
Vibrio alginolyticus
Listonella anguillara
Vibrio harveyi
Vibriosis
Aquaculture
description This work was funded by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within the scope of the project MARE (UIDB/04292/2020 and UIDP/04292/2020) and grant agreement n. POCI-01-0247-FEDER-035234; AlgaValor, Portugal 2020 program.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022-01-01T00:00:00Z
2023-07-28T15:12:57Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.8/8708
url http://hdl.handle.net/10400.8/8708
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Costa, C.; Ferreira, G.D.; Simões, M.; Silva, J.L.; Campos, M.J. Real-Time PCR Protocol for Detection and Quantification of Three Pathogenic Members of the Vibrionaceae Family. Microorganisms 2022, 10, 2060. https://doi.org/ 10.3390/microorganisms10102060
2076-2607
10.3390/microorganisms10102060
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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