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An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer
BACKGROUND: An accurate genotyping analysis is one of the critical prerequisites for patients with colorectal cancer receiving matched therapies. Conventional genotyping analysis is currently used to detect either gene mutations or MSI status, delaying the detection of critical tumor biomarkers and...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10134345/ https://www.ncbi.nlm.nih.gov/pubmed/36583506 http://dx.doi.org/10.1002/cam4.5557 |
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author | Liang, Li Li, Xin Nong, Lin Cai, Weijing Zhang, Jixin Liu, Ping Li, Ting |
author_facet | Liang, Li Li, Xin Nong, Lin Cai, Weijing Zhang, Jixin Liu, Ping Li, Ting |
author_sort | Liang, Li |
collection | PubMed |
description | BACKGROUND: An accurate genotyping analysis is one of the critical prerequisites for patients with colorectal cancer receiving matched therapies. Conventional genotyping analysis is currently used to detect either gene mutations or MSI status, delaying the detection of critical tumor biomarkers and thus the optimal time for treatment. An assay that analyzes both biomarkers in a streamlined process is eagerly needed. METHODS: We developed an assay combining Multiplex PCR Amplification, Single‐base Extension and capillary electrophoresis (CE) analysis (MASE‐CE) for synchronous detection of KRAS/NRAS/BRAF mutations and MSI status. In a 190 colorectal cancer cohort, we identified seven somatic mutations in KRAS, NRAS and BRAF as well as five MSI loci (D2S123/D5S346/D17S250/BAT‐25/BAT‐26) simultaneously. KRAS/NRAS/BRAF mutations were detected by NGS and MASE‐CE, and MSI status were detected by PCR‐CE and MASE‐CE methods. RESULTS: The MASE‐CE method showed high consistency with NGS for mutation detection (Kappa value ≥0.8) and PCR‐CE (Kappa value = 0.79). In addition, the limits of detection (LOD) of MASE‐CE assay for MSI and somatic mutation were 5% and 2%, respectively. CONCLUSIONS: In somatic mutation detection and MSI detection, the LOD of MASE‐CE assay was superior to that of qPCR and NGS. MASE‐CE assay is a highly sensitive, time‐saving and specimen‐saving method, which can greatly avoid the cumbersome testing process and provide clinical decision for doctors in time. |
format | Online Article Text |
id | pubmed-10134345 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101343452023-04-28 An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer Liang, Li Li, Xin Nong, Lin Cai, Weijing Zhang, Jixin Liu, Ping Li, Ting Cancer Med RESEARCH ARTICLES BACKGROUND: An accurate genotyping analysis is one of the critical prerequisites for patients with colorectal cancer receiving matched therapies. Conventional genotyping analysis is currently used to detect either gene mutations or MSI status, delaying the detection of critical tumor biomarkers and thus the optimal time for treatment. An assay that analyzes both biomarkers in a streamlined process is eagerly needed. METHODS: We developed an assay combining Multiplex PCR Amplification, Single‐base Extension and capillary electrophoresis (CE) analysis (MASE‐CE) for synchronous detection of KRAS/NRAS/BRAF mutations and MSI status. In a 190 colorectal cancer cohort, we identified seven somatic mutations in KRAS, NRAS and BRAF as well as five MSI loci (D2S123/D5S346/D17S250/BAT‐25/BAT‐26) simultaneously. KRAS/NRAS/BRAF mutations were detected by NGS and MASE‐CE, and MSI status were detected by PCR‐CE and MASE‐CE methods. RESULTS: The MASE‐CE method showed high consistency with NGS for mutation detection (Kappa value ≥0.8) and PCR‐CE (Kappa value = 0.79). In addition, the limits of detection (LOD) of MASE‐CE assay for MSI and somatic mutation were 5% and 2%, respectively. CONCLUSIONS: In somatic mutation detection and MSI detection, the LOD of MASE‐CE assay was superior to that of qPCR and NGS. MASE‐CE assay is a highly sensitive, time‐saving and specimen‐saving method, which can greatly avoid the cumbersome testing process and provide clinical decision for doctors in time. John Wiley and Sons Inc. 2022-12-30 /pmc/articles/PMC10134345/ /pubmed/36583506 http://dx.doi.org/10.1002/cam4.5557 Text en © 2022 The Authors. Cancer Medicine published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | RESEARCH ARTICLES Liang, Li Li, Xin Nong, Lin Cai, Weijing Zhang, Jixin Liu, Ping Li, Ting An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title | An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title_full | An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title_fullStr | An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title_full_unstemmed | An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title_short | An innovative single‐base extension method for synchronous detection of point mutations and MSI status in colorectal cancer |
title_sort | innovative single‐base extension method for synchronous detection of point mutations and msi status in colorectal cancer |
topic | RESEARCH ARTICLES |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10134345/ https://www.ncbi.nlm.nih.gov/pubmed/36583506 http://dx.doi.org/10.1002/cam4.5557 |
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