N_LyST: a simple and rapid screening test for Lynch Syndrome
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 mononucleoti...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
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BMJ Publishing Group Ltd
2018
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| Online Access: | https://eprints.nottingham.ac.uk/49611/ |
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| author | Susanti, Susanti Fadhil, Wakkas Ebili, Henry O. 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 |
| author_facet | Susanti, Susanti Fadhil, Wakkas Ebili, Henry O. 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 |
| author_sort | Susanti, Susanti |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T20:13:24Z |
| format | Article |
| id | nottingham-49611 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:13:24Z |
| publishDate | 2018 |
| publisher | BMJ Publishing Group Ltd |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-496112020-05-04T19:33:23Z https://eprints.nottingham.ac.uk/49611/ N_LyST: a simple and rapid screening test for Lynch Syndrome Susanti, Susanti Fadhil, Wakkas Ebili, Henry O. 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 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. BMJ Publishing Group Ltd 2018-08-31 Article PeerReviewed Susanti, Susanti, Fadhil, Wakkas, Ebili, Henry O., Asiri, Abutaleb, Nestarenkaite, Ausrine, Hadjimichael, Efthymios, Ham-Karim, Hersh A., Field, Joanne, Stafford, Katherine, Matharoo-Ball, Balwir, Hassall, James C., Sharif, Abid, Oniscu, Anca and Ilyas, Mohammad (2018) N_LyST: a simple and rapid screening test for Lynch Syndrome. Journal of Clinical Pathology, 71 (8). pp. 713-720. ISSN 1472-4146 Microsatellite Instability (MSI); High Resolution Melting (HRM); Colorectal cancer; Lynch Syndrome; BRAF mutation; Promoter methylation http://jcp.bmj.com/content/early/2018/02/21/jclinpath-2018-205013 doi:10.1136/jclinpath-2018-205013 doi:10.1136/jclinpath-2018-205013 |
| spellingShingle | Microsatellite Instability (MSI); High Resolution Melting (HRM); Colorectal cancer; Lynch Syndrome; BRAF mutation; Promoter methylation Susanti, Susanti Fadhil, Wakkas Ebili, Henry O. 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 |
| title | N_LyST: a simple and rapid screening test for Lynch Syndrome |
| title_full | N_LyST: a simple and rapid screening test for Lynch Syndrome |
| title_fullStr | N_LyST: a simple and rapid screening test for Lynch Syndrome |
| title_full_unstemmed | N_LyST: a simple and rapid screening test for Lynch Syndrome |
| title_short | N_LyST: a simple and rapid screening test for Lynch Syndrome |
| title_sort | n_lyst: a simple and rapid screening test for lynch syndrome |
| topic | Microsatellite Instability (MSI); High Resolution Melting (HRM); Colorectal cancer; Lynch Syndrome; BRAF mutation; Promoter methylation |
| url | https://eprints.nottingham.ac.uk/49611/ https://eprints.nottingham.ac.uk/49611/ https://eprints.nottingham.ac.uk/49611/ |