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N_LyST: a simple and rapid screening test for Lynch Syndrome

Susanti, Susanti; Fadhil, Wakkas; Ebili, Henry Okuchukwu; Asiri, Abutaleb; Nestarenkaite, Ausrine; Hadjimichael, Efthymios; Ham-Karim, Hersh A.; Field, Joanne; Stafford, Katherine; Matharoo-Ball, Balwir; Hassall, James C.; Sharif, Abid; Oniscu, Anca; Ilyas, Mohammad

N_LyST: a simple and rapid screening test for Lynch Syndrome Thumbnail


Authors

Susanti Susanti

Wakkas Fadhil

Henry Okuchukwu Ebili

Abutaleb Asiri

Ausrine Nestarenkaite

Efthymios Hadjimichael

Hersh A. Ham-Karim

Joanne Field

Katherine Stafford

Balwir Matharoo-Ball

James C. Hassall

Abid Sharif

Anca Oniscu



Abstract

Aims: We sought to use PCR followed by high-resolution melting (HRM) analysis to develop a single closed-tube screening panel to screen for Lynch Syndrome. This comprises tests for microsatellite instability (MSI), MLH1 methylation promoter and BRAF mutation.
Methods:For MSI-testing, 5 mononucleotide markers (BAT25, BAT26, BCAT25, MYB, EWSR1) were developed. In addition, primers were designed to interrogate Region C of the MLH1 promoter for methylation (using bisulphite-modified DNA) and to test for mutations in codon 600 of BRAF. Two separate cohorts from Nottingham (n = 99, 46 with MSI, 53 being microsatellite stable (MSS)) and Edinburgh (n=88, 45 MSI, 43 MSS).
Results:All the cases (n=187) were blind tested for MSI and all were correctly characterised by our panel. The MLH1 promoter and BRAF were tested only in the Nottingham cohort. Successful blinded analysis was performed on the MLH1 promoter in 97 cases. All MSS cases showed a pattern of non-methylation whilst 41/44 cases with MSI showed full methylation. The three cases with MSI and a non-methylated pattern had aberrations in MSH2 and MSH6 expression. BRAF mutation was detected in 61% of MSI cases and 11% of MSS cases.
Finally, 12 cases were blind screened by using the whole panel as a single test. Of these, 5 were identified as MSS, 4 as MSI/non-LS and 3 as MSI/possible LS. These results were concordant with the previous data.
Conclusion: We describe the Nottingham Lynch Syndrome Test (N_LyST). This is a quick simple cheap method for screening for Lynch Syndrome.

Citation

Susanti, S., Fadhil, W., Ebili, H. O., Asiri, A., Nestarenkaite, A., Hadjimichael, E., Ham-Karim, H. A., Field, J., Stafford, K., Matharoo-Ball, B., Hassall, J. C., Sharif, A., Oniscu, A., & Ilyas, M. (2018). N_LyST: a simple and rapid screening test for Lynch Syndrome. Journal of Clinical Pathology, 71(8), 713-720. https://doi.org/10.1136/jclinpath-2018-205013

Journal Article Type Article
Acceptance Date Feb 2, 2018
Online Publication Date Feb 22, 2018
Publication Date Aug 31, 2018
Deposit Date Feb 7, 2018
Publicly Available Date Feb 22, 2018
Journal Journal of Clinical Pathology
Print ISSN 0021-9746
Electronic ISSN 1472-4146
Publisher BMJ Publishing Group
Peer Reviewed Peer Reviewed
Volume 71
Issue 8
Pages 713-720
DOI https://doi.org/10.1136/jclinpath-2018-205013
Keywords Microsatellite Instability (MSI); High Resolution Melting (HRM); Colorectal cancer; Lynch Syndrome; BRAF mutation; Promoter methylation
Public URL https://nottingham-repository.worktribe.com/output/913033
Publisher URL https://jcp.bmj.com/content/71/8/713
Additional Information © 2018 BMJ Publishing Group Ltd & Association of Clinical Pathologists
Contract Date Feb 7, 2018