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Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography

High-performance liquid chromatography (HPLC) is a recently introduced high-capacity automated method for detecting unknown mutations (denaturing HPLC) or for sizing DNA fragments under nondenaturing conditions. We have adapted the HPLC method for detection of loss of heterozygosity (LOH) and used g...

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Autores principales: Chernova, O B, Barnett, G H, Cowell, J K
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2003
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2741124/
https://www.ncbi.nlm.nih.gov/pubmed/12799632
http://dx.doi.org/10.1038/sj.bjc.6601025
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author Chernova, O B
Barnett, G H
Cowell, J K
author_facet Chernova, O B
Barnett, G H
Cowell, J K
author_sort Chernova, O B
collection PubMed
description High-performance liquid chromatography (HPLC) is a recently introduced high-capacity automated method for detecting unknown mutations (denaturing HPLC) or for sizing DNA fragments under nondenaturing conditions. We have adapted the HPLC method for detection of loss of heterozygosity (LOH) and used glial tumours as a model to evaluate its sensitivity and specificity in comparison to conventional denaturing polyacrylamide gel electrophoresis. A total of 20 oligodendrogliomas (grades II and III), and five astrocytic tumours (grades III and IV) were analysed using 14 polymorphic microsatellite markers mapping to regions on chromosomes 1p, 19q, and 10q using both DNA-HPLC and denaturing gel electrophoresis. This study demonstrated complete concordance between both methods. However, unlike gel electrophoresis, HPLC is automated, does not require post-PCR processing, and does not require hazardous radioactive or expensive fluorescent labelling. Our data suggest that HPLC is a reliable and sensitive method for detection of allelic losses in tumour samples and it is a favourable alternative for high-sensitivity LOH detection in both research and diagnostic environments.
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spelling pubmed-27411242009-09-10 Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography Chernova, O B Barnett, G H Cowell, J K Br J Cancer Molecular and Cellular Pathology High-performance liquid chromatography (HPLC) is a recently introduced high-capacity automated method for detecting unknown mutations (denaturing HPLC) or for sizing DNA fragments under nondenaturing conditions. We have adapted the HPLC method for detection of loss of heterozygosity (LOH) and used glial tumours as a model to evaluate its sensitivity and specificity in comparison to conventional denaturing polyacrylamide gel electrophoresis. A total of 20 oligodendrogliomas (grades II and III), and five astrocytic tumours (grades III and IV) were analysed using 14 polymorphic microsatellite markers mapping to regions on chromosomes 1p, 19q, and 10q using both DNA-HPLC and denaturing gel electrophoresis. This study demonstrated complete concordance between both methods. However, unlike gel electrophoresis, HPLC is automated, does not require post-PCR processing, and does not require hazardous radioactive or expensive fluorescent labelling. Our data suggest that HPLC is a reliable and sensitive method for detection of allelic losses in tumour samples and it is a favourable alternative for high-sensitivity LOH detection in both research and diagnostic environments. Nature Publishing Group 2003-06-16 2003-06-10 /pmc/articles/PMC2741124/ /pubmed/12799632 http://dx.doi.org/10.1038/sj.bjc.6601025 Text en Copyright © 2003 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Molecular and Cellular Pathology
Chernova, O B
Barnett, G H
Cowell, J K
Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title_full Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title_fullStr Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title_full_unstemmed Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title_short Rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
title_sort rapid detection of allelic losses in brain tumours using microsatellite repeat markers and high-performance liquid chromatography
topic Molecular and Cellular Pathology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2741124/
https://www.ncbi.nlm.nih.gov/pubmed/12799632
http://dx.doi.org/10.1038/sj.bjc.6601025
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