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Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples

2023-06Artigo científico Acesso aberto
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dc.contributor.authorOliveira, Beatriz B.
dc.contributor.authorCosta, Beatriz
dc.contributor.authorMorão, Bárbara
dc.contributor.authorFaias, Sandra
dc.contributor.authorVeigas, Bruno
dc.contributor.authorPereira, Lucília Pebre
dc.contributor.authorAlbuquerque, Cristina
dc.contributor.authorMaio, Rui
dc.contributor.authorCravo, Marília
dc.contributor.authorFernandes, Alexandra R.
dc.contributor.authorBaptista, Pedro Viana
dc.contributor.institutionDCV - Departamento de Ciências da Vida
dc.contributor.institutionUCIBIO - Applied Molecular Biosciences Unit
dc.contributor.institutionNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
dc.contributor.pblSpringer
dc.date.accessioned2023-07-04T22:18:07Z
dc.date.available2023-07-04T22:18:07Z
dc.date.issued2023-06
dc.descriptionPublisher Copyright: © 2023, The Author(s).
dc.description.abstractThe success of personalized medicine depends on the discovery of biomarkers that allow oncologists to identify patients that will benefit from a particular targeted drug. Molecular tests are mostly performed using tumor samples, which may not be representative of the tumor’s temporal and spatial heterogeneity. Liquid biopsies, and particularly the analysis of circulating tumor DNA, are emerging as an interesting means for diagnosis, prognosis, and predictive biomarker discovery. In this study, the amplification refractory mutation system (ARMS) coupled with high-resolution melting analysis (HRMA) was developed for detecting two of the most relevant KRAS mutations in codon 12. After optimization with commercial cancer cell lines, KRAS mutation screening was validated in tumor and plasma samples collected from patients with pancreatic ductal adenocarcinoma (PDAC), and the results were compared to those obtained by Sanger sequencing (SS) and droplet digital polymerase chain reaction (ddPCR). The developed ARMS-HRMA methodology stands out for its simplicity and reduced time to result when compared to both SS and ddPCR but showing high sensitivity and specificity for the detection of mutations in tumor and plasma samples. In fact, ARMS-HRMA scored 3 more mutations compared to SS (tumor samples T6, T7, and T12) and one more compared to ddPCR (tumor sample T7) in DNA extracted from tumors. For ctDNA from plasma samples, insufficient genetic material prevented the screening of all samples. Still, ARMS-HRMA allowed for scoring more mutations in comparison to SS and 1 more mutation in comparison to ddPCR (plasma sample P7). We propose that ARMS-HRMA might be used as a sensitive, specific, and simple method for the screening of low-level mutations in liquid biopsies, suitable for improving diagnosis and prognosis schemes. Graphical Abstract: [Figure not available: see fulltext.]en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent15
dc.format.extent4060695
dc.identifier.doi10.1007/s00216-023-04696-6
dc.identifier.issn1618-2642
dc.identifier.otherPURE: 65212596
dc.identifier.otherPURE UUID: b2c4ea4d-1977-452d-ab83-a93d551434ce
dc.identifier.otherScopus: 85153507668
dc.identifier.otherWOS: 000978928300001
dc.identifier.otherPubMed: 37097304
dc.identifier.otherPubMedCentral: PMC10185647
dc.identifier.otherORCID: /0000-0001-5255-7095/work/151383877
dc.identifier.otherORCID: /0000-0003-2054-4438/work/151392209
dc.identifier.urihttp://hdl.handle.net/10362/154848
dc.identifier.urlhttps://www.scopus.com/pages/publications/85153507668
dc.language.isoeng
dc.peerreviewedyes
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
dc.relationApplied Molecular Biosciences Unit
dc.relationApplied Molecular Biosciences Unit
dc.relationInstitute for Health and Bioeconomy
dc.relationSimplified chip prototyping for improved screening of gene-silencing therapeutics using 3D cell models.
dc.relationinfo:eu-repo/grantAgreement/FCT/OE/2020.07660.BD/PT
dc.subjectARMS-HRMA
dc.subjectCirculating tumor DNA
dc.subjectMutation detection
dc.subjectPancreatic cancer
dc.subjectAnalytical Chemistry
dc.subjectBiochemistry
dc.subjectSDG 3 - Good Health and Well-being
dc.titleCombining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samplesen
dc.typejournal article
degois.publication.firstPage2849
degois.publication.issue14
degois.publication.lastPage2863
degois.publication.titleAnalytical and Bioanalytical Chemistry
degois.publication.volume415
dspace.entity.typePublication
oaire.awardNumberUIDP/04378/2020
oaire.awardNumberUIDB/04378/2020
oaire.awardNumberLA/P/0140/2020
oaire.awardNumber2020.07660.BD
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleInstitute for Health and Bioeconomy
oaire.awardTitleSimplified chip prototyping for improved screening of gene-silencing therapeutics using 3D cell models.
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0140%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/OE/2020.07660.BD/PT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStreamOE
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
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
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccess
relation.isProjectOfPublication38373452-5c9c-4724-8def-f03314ecce0e
relation.isProjectOfPublicatione07cf232-4705-4b5b-b2c4-af8f25311076
relation.isProjectOfPublication834c369e-6900-4105-aa78-6ba0e52f96ca
relation.isProjectOfPublication66916e71-00c7-4cb8-b4bc-c2e81ab9d2a6
relation.isProjectOfPublication.latestForDiscovery38373452-5c9c-4724-8def-f03314ecce0e

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