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/10451/57280 |
Resumo: | © The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
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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 cancer© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.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.This work is financed by national funds from FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. FCT-MCTES is also acknowledged for 2020.07660.BD for BBO. Open access funding provided by FCT|FCCN (b-on).Springer NatureRepositório da Universidade de LisboaOliveira, Beatriz B.Costa, BeatrizMorão, BarbaraFaias, SandraVeigas, BrunoPereira, Lucília PebreAlbuquerque, CristinaMaio, RuiCravo, MaríliaFernandes, Alexandra R.Baptista, Pedro Viana2023-04-27T14:18:43Z20232023-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/57280engAnal Bioanal Chem. 2023 Apr 251618-264210.1007/s00216-023-04696-61618-2650info: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:RCAAP2024-11-20T18:20:56Zoai:repositorio.ul.pt:10451/57280Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-11-20T18:20:56Repositório 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 |
| 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, Barbara 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, Barbara 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 |
Repositório da Universidade de Lisboa |
| dc.contributor.author.fl_str_mv |
Oliveira, Beatriz B. Costa, Beatriz Morão, Barbara 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 |
| topic |
ARMS-HRMA Circulating tumor DNA Mutation detection Pancreatic cancer |
| description |
© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
| publishDate |
2023 |
| dc.date.none.fl_str_mv |
2023-04-27T14:18:43Z 2023 2023-01-01T00:00:00Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
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http://hdl.handle.net/10451/57280 |
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http://hdl.handle.net/10451/57280 |
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eng |
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eng |
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Anal Bioanal Chem. 2023 Apr 25 1618-2642 10.1007/s00216-023-04696-6 1618-2650 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Springer Nature |
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Springer Nature |
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