Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2023 |
| Outros Autores: | , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/154848 |
Resumo: | Publisher Copyright: © 2023, The Author(s). |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samplesARMS-HRMACirculating tumor DNAMutation detectionPancreatic cancerAnalytical ChemistryBiochemistrySDG 3 - Good Health and Well-beingPublisher Copyright: © 2023, The Author(s).The 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.]DCV - Departamento de Ciências da VidaUCIBIO - Applied Molecular Biosciences UnitNOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)RUNOliveira, Beatriz B.Costa, BeatrizMorão, BárbaraFaias, SandraVeigas, BrunoPereira, Lucília PebreAlbuquerque, CristinaMaio, RuiCravo, MaríliaFernandes, Alexandra R.Baptista, Pedro Viana2023-07-04T22:18:07Z2023-062023-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article15application/pdfhttp://hdl.handle.net/10362/154848eng1618-2642PURE: 65212596https://doi.org/10.1007/s00216-023-04696-6info: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:RCAAP2024-03-11T05:37:18Zoai:run.unl.pt:10362/154848Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:55:47.555547Repositório 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 |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| title |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| spellingShingle |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples Oliveira, Beatriz B. ARMS-HRMA Circulating tumor DNA Mutation detection Pancreatic cancer Analytical Chemistry Biochemistry SDG 3 - Good Health and Well-being |
| title_short |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| title_full |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| title_fullStr |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| title_full_unstemmed |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| title_sort |
Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples |
| author |
Oliveira, Beatriz B. |
| author_facet |
Oliveira, Beatriz B. Costa, Beatriz Morão, Bárbara Faias, Sandra Veigas, Bruno Pereira, Lucília Pebre Albuquerque, Cristina Maio, Rui Cravo, Marília Fernandes, Alexandra R. Baptista, Pedro Viana |
| author_role |
author |
| author2 |
Costa, Beatriz Morão, Bárbara Faias, Sandra Veigas, Bruno Pereira, Lucília Pebre Albuquerque, Cristina Maio, Rui Cravo, Marília Fernandes, Alexandra R. Baptista, Pedro Viana |
| author2_role |
author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
DCV - Departamento de Ciências da Vida UCIBIO - Applied Molecular Biosciences Unit NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) RUN |
| dc.contributor.author.fl_str_mv |
Oliveira, Beatriz B. Costa, Beatriz Morão, Bárbara Faias, Sandra Veigas, Bruno Pereira, Lucília Pebre Albuquerque, Cristina Maio, Rui Cravo, Marília Fernandes, Alexandra R. Baptista, Pedro Viana |
| dc.subject.por.fl_str_mv |
ARMS-HRMA Circulating tumor DNA Mutation detection Pancreatic cancer Analytical Chemistry Biochemistry SDG 3 - Good Health and Well-being |
| topic |
ARMS-HRMA Circulating tumor DNA Mutation detection Pancreatic cancer Analytical Chemistry Biochemistry SDG 3 - Good Health and Well-being |
| description |
Publisher Copyright: © 2023, The Author(s). |
| publishDate |
2023 |
| dc.date.none.fl_str_mv |
2023-07-04T22:18:07Z 2023-06 2023-06-01T00:00:00Z |
| 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/10362/154848 |
| url |
http://hdl.handle.net/10362/154848 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
1618-2642 PURE: 65212596 https://doi.org/10.1007/s00216-023-04696-6 |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
| dc.format.none.fl_str_mv |
15 application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799138144402014208 |