Academic Journal
Molecular testing for familial hypercholesterolaemia-associated mutations in a UK-based cohort: development of an NGS-based method and comparison with multiplex polymerase chain reaction and oligonucleotide arrays
| العنوان: | Molecular testing for familial hypercholesterolaemia-associated mutations in a UK-based cohort: development of an NGS-based method and comparison with multiplex polymerase chain reaction and oligonucleotide arrays |
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| المؤلفون: | Reiman, Anne, Pandey, Sarojini, Lloyd, Kate L, Dyer, Nigel, Khan, Mike, Crockard, Martin, Latten, Mark J, Watson, Tracey L, Cree, Ian A, Grammatopoulos, Dimitris K |
| المصدر: | Annals of Clinical Biochemistry: International Journal of Laboratory Medicine ; volume 53, issue 6, page 654-662 ; ISSN 0004-5632 1758-1001 |
| بيانات النشر: | SAGE Publications |
| سنة النشر: | 2016 |
| الوصف: | Background Detection of disease-associated mutations in patients with familial hypercholesterolaemia is crucial for early interventions to reduce risk of cardiovascular disease. Screening for these mutations represents a methodological challenge since more than 1200 different causal mutations in the low-density lipoprotein receptor has been identified. A number of methodological approaches have been developed for screening by clinical diagnostic laboratories. Methods Using primers targeting, the low-density lipoprotein receptor, apolipoprotein B, and proprotein convertase subtilisin/kexin type 9, we developed a novel Ion Torrent-based targeted re-sequencing method. We validated this in a West Midlands-UK small cohort of 58 patients screened in parallel with other mutation-targeting methods, such as multiplex polymerase chain reaction (Elucigene FH20), oligonucleotide arrays (Randox familial hypercholesterolaemia array) or the Illumina next-generation sequencing platform. Results In this small cohort, the next-generation sequencing method achieved excellent analytical performance characteristics and showed 100% and 89% concordance with the Randox array and the Elucigene FH20 assay. Investigation of the discrepant results identified two cases of mutation misclassification of the Elucigene FH20 multiplex polymerase chain reaction assay. A number of novel mutations not previously reported were also identified by the next-generation sequencing method. Conclusions Ion Torrent-based next-generation sequencing can deliver a suitable alternative for the molecular investigation of familial hypercholesterolaemia patients, especially when comprehensive mutation screening for rare or unknown mutations is required. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1177/0004563216629170 |
| الاتاحة: | https://doi.org/10.1177/0004563216629170 https://journals.sagepub.com/doi/pdf/10.1177/0004563216629170 https://journals.sagepub.com/doi/full-xml/10.1177/0004563216629170 |
| Rights: | https://journals.sagepub.com/page/policies/text-and-data-mining-license |
| رقم الانضمام: | edsbas.8996F2B1 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1177/0004563216629170 |
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