Repository logo
 
Publication

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

2022Journal article Open access
Simple item page
dc.contributor.authorCosta, Cátia
dc.contributor.authorFerreira, Guilherme D.
dc.contributor.authorSimões, Marco
dc.contributor.authorSilva, Joana L.
dc.contributor.authorCampos, Maria J.
dc.date.accessioned2023-07-28T15:12:57Z
dc.date.available2023-07-28T15:12:57Z
dc.date.issued2022
dc.descriptionThis 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.pt_PT
dc.description.abstractVibriosis, 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.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationCosta, 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/microorganisms10102060pt_PT
dc.identifier.doi10.3390/microorganisms10102060pt_PT
dc.identifier.issn2076-2607
dc.identifier.urihttp://hdl.handle.net/10400.8/8708
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMDPIpt_PT
dc.relationMarine and Environmental Sciences Centre
dc.relationMarine and Environmental Sciences Centre
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectqPCRpt_PT
dc.subjectVibrio alginolyticuspt_PT
dc.subjectListonella anguillarapt_PT
dc.subjectVibrio harveyipt_PT
dc.subjectVibriosispt_PT
dc.subjectAquaculturept_PT
dc.titleReal-time PCR protocol for detection and quantification of three pathogenic members of the Vibrionaceae familypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleMarine and Environmental Sciences Centre
oaire.awardTitleMarine and Environmental Sciences Centre
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04292%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04292%2F2020/PT
oaire.citation.issue10pt_PT
oaire.citation.titleMicroorganismspt_PT
oaire.citation.volume10pt_PT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
person.familyNameGaspar Veríssimo Costa
person.familyNameDuarte Ferreira
person.familyNameDias Simões
person.familyNameCampos
person.givenNameCátia Filipa
person.givenNameGuilherme
person.givenNameMarco Aurélio
person.givenNameMaria Jorge
person.identifierC-2471-2018
person.identifier174191
person.identifier.ciencia-idB61B-C0B9-89DF
person.identifier.ciencia-idD017-9023-9519
person.identifier.ciencia-id1D1C-C217-F68A
person.identifier.ciencia-id2416-7867-2D40
person.identifier.orcid0000-0002-8558-5786
person.identifier.orcid0000-0003-0010-4558
person.identifier.orcid0000-0001-7995-7304
person.identifier.orcid0000-0002-0883-0610
person.identifier.ridC-2471-2018
person.identifier.ridE-5591-2014
person.identifier.scopus-author-id57211559216
person.identifier.scopus-author-id55767069800
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication0d8da2af-5614-4a5e-9772-46aeebf02767
relation.isAuthorOfPublication8823b64b-9770-487c-b1fd-c28fd3924649
relation.isAuthorOfPublicatione2a25105-22d0-4e1d-9e51-2befd64f1baf
relation.isAuthorOfPublication6d1bf3db-fdba-4bea-b65d-94491f841659
relation.isAuthorOfPublication.latestForDiscoverye2a25105-22d0-4e1d-9e51-2befd64f1baf
relation.isProjectOfPublication54154db1-7a14-4ce1-b54e-b3a01fcd44f5
relation.isProjectOfPublicationa66a15eb-ff92-44d8-a34e-0f87f0c01d01
relation.isProjectOfPublication.latestForDiscovery54154db1-7a14-4ce1-b54e-b3a01fcd44f5

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Costa et al_2022_microorganisms.pdf
Size:
3.08 MB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.32 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

MARE - Artigos com peer review
ESTM - Artigos em revistas internacionais