A Novel Colorectal Cancer Test Combining Microsatellite Instability and Analysis: Clinical Validation and Impact on Lynch Syndrome Screening

Overview

Journal BJC Rep

Publisher Nature Portfolio

Date 2024 Jul 4

PMID 38962168

Authors

Richard Gallon

Patricia Herrero-Belmonte

Rachel Phelps

Christine Hayes

Elizabeth Sollars

Daniel Egan

Helena Spiewak

Sam Nalty

Sarah Mills

Peh Sun Loo

Gillian M Borthwick

Mauro Santibanez-Koref

John Burn

Ciaron McAnulty

Michael S Jackson

Affiliations

Soon will be listed here.

Abstract

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 , , and mutation hotspots. MSI and 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.

Citing Articles

Detection of Mismatch Repair Deficiency in Endometrial Cancer: Assessment of IHC, Fragment Length Analysis, and Amplicon Sequencing Based MSI Testing.

Sowter P, Gallon R, Hayes C, Phelps R, Borthwick G, Prior S Cancers (Basel). 2024; 16(23).

PMID: 39682157 PMC: 11640224. DOI: 10.3390/cancers16233970.

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