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Lookup NU author(s): Dr Richard Gallon, Rachel Phelps, Christine Hayes, Sarah Mills, Dr Gillian Borthwick, Dr Mauro Santibanez Koref, Dr Ciaron McAnulty, Dr Michael Jackson
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Background: Lynch syndrome (LS) is under-diagnosed. UK National Institute for Health and Care Excellence guidelines recommend multistep molecular testing of all colorectal cancers (CRCs) to screen for LS. However, the complexity of the pathway has resulted in limited improvement in diagnosis.Methods: One-step multiplex PCR was used to generate sequencing-ready amplicons from 14 microsatellite instability (MSI) markers and 22 BRAF, KRAS, and NRAS mutation hotspots. MSI and BRAF/RAS variants were detected using amplicon-sequencing and automated analysis. The assay was clinically validated and deployed into service in northern England, followed by regional and local audits to assess its impact.Results: MSI analysis achieved 99.1% sensitivity and 99.2% specificity and was reproducible (r = 0.995). Mutation hotspot analysis had 100% sensitivity, 99.9% specificity, and was reproducible (r = 0.998). Assay-use in service in 2022-2023 increased CRC testing (97.2% (2466/2536) versus 28.6% (601/2104)), halved turnaround times, and identified more CRC patients at-risk of LS (5.5% (139/2536) versus 2.9% (61/2104)) compared to 2019-2020 when a multi-test pathway was used.Conclusion: A novel amplicon-sequencing assay of CRCs, including all biomarkers for LS screening and anti-EGFR therapy, achieved >95% testing rate. Adoption of this low cost, scalable, and fully automatable test will complement on-going, national initiatives to improve LS screening.
Author(s): Gallon R, Herrero-Belmonte P, Phelps R, Hayes C, Sollars E, Egan D, Spiewak H, Nalty S, Mills S, Loo P-S, Borthwick G, Santibanez-Koref M, Burn J, McAnulty C, Jackson MS
Publication type: Article
Publication status: Published
Journal: BJC reports
Year: 2024
Volume: 2
Issue: 1
Online publication date: 01/07/2024
Acceptance date: 09/06/2024
Date deposited: 20/11/2024
ISSN (electronic): 2731-9377
Publisher: Nature
URL: https://doi.org/10.1038/s44276-024-00072-8
DOI: 10.1038/s44276-024-00072-8
Data Access Statement: Amplicon sequence FASTQ files are available from the European Nucleotide Archive (https://www.ebi.ac.uk/ena/browser/home) using study ID PRJEB67820. R (version 4.2.2) code for simulation analyses is available on request from the corresponding author.
PubMed id: 38962168
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