PURPOSE: Familial hypercholesterolemia is a common Mendelian disorder associated with early-onset coronary heart disease that can be treated by cholesterol-lowering drugs. The majority of cases in the United Kingdom are currently without a molecular diagnosis, which is partly due to the cost and time associated with standard screening techniques. The main purpose of this study was to test the sensitivity and specificity of two next-generation sequencing protocols for genetic diagnosis of familial hypercholesterolemia. METHODS: Libraries were prepared for next-generation sequencing by two target enrichment protocols; one using the SureSelect Target Enrichment System and the other using the PCR-based Access Array platform. RESULTS: In the validation cohort, both protocols showed 100% specificity, whereas the sensitivity for short variant detection was 100% for the SureSelect Target Enrichment and 98% for the Access Array protocol. Large deletions/duplications were only detected using the SureSelect Target Enrichment protocol. In the prospective cohort, the mutation detection rate using the Access Array was highest in patients with clinically definite familial hypercholesterolemia (67%), followed by patients with possible familial hypercholesterolemia (26%). CONCLUSION: We have shown the potential of target enrichment methods combined with next-generation sequencing for molecular diagnosis of familial hypercholesterolemia. Adopting these assays for patients with suspected familial hypercholesterolemia could improve cost-effectiveness and increase the overall number of patients with a molecular diagnosis.
PURPOSE: Familial hypercholesterolemia is a common Mendelian disorder associated with early-onset coronary heart disease that can be treated by cholesterol-lowering drugs. The majority of cases in the United Kingdom are currently without a molecular diagnosis, which is partly due to the cost and time associated with standard screening techniques. The main purpose of this study was to test the sensitivity and specificity of two next-generation sequencing protocols for genetic diagnosis of familial hypercholesterolemia. METHODS: Libraries were prepared for next-generation sequencing by two target enrichment protocols; one using the SureSelect Target Enrichment System and the other using the PCR-based Access Array platform. RESULTS: In the validation cohort, both protocols showed 100% specificity, whereas the sensitivity for short variant detection was 100% for the SureSelect Target Enrichment and 98% for the Access Array protocol. Large deletions/duplications were only detected using the SureSelect Target Enrichment protocol. In the prospective cohort, the mutation detection rate using the Access Array was highest in patients with clinically definite familial hypercholesterolemia (67%), followed by patients with possible familial hypercholesterolemia (26%). CONCLUSION: We have shown the potential of target enrichment methods combined with next-generation sequencing for molecular diagnosis of familial hypercholesterolemia. Adopting these assays for patients with suspected familial hypercholesterolemia could improve cost-effectiveness and increase the overall number of patients with a molecular diagnosis.
Authors: Catherine E Cottrell; Hussam Al-Kateb; Andrew J Bredemeyer; Eric J Duncavage; David H Spencer; Haley J Abel; Christina M Lockwood; Ian S Hagemann; Stephanie M O'Guin; Lauren C Burcea; Christopher S Sawyer; Dayna M Oschwald; Jennifer L Stratman; Dorie A Sher; Mark R Johnson; Justin T Brown; Paul F Cliften; Bijoy George; Leslie D McIntosh; Savita Shrivastava; Tudung T Nguyen; Jacqueline E Payton; Mark A Watson; Seth D Crosby; Richard D Head; Robi D Mitra; Rakesh Nagarajan; Shashikant Kulkarni; Karen Seibert; Herbert W Virgin; Jeffrey Milbrandt; John D Pfeifer Journal: J Mol Diagn Date: 2013-11-06 Impact factor: 5.568
Authors: P Cacheiro; A Ordóñez-Ugalde; B Quintáns; S Piñeiro-Hermida; J Amigo; M García-Murias; S I Pascual-Pascual; F Grandas; J Arpa; A Carracedo; M J Sobrido Journal: Mol Diagn Ther Date: 2017-06 Impact factor: 4.074
Authors: Christopher Carroll; Paul Tappenden; Rachid Rafia; Jean Hamilton; Duncan Chambers; Mark Clowes; Paul Durrington; Nadeem Qureshi; Anthony S Wierzbicki Journal: Pharmacoeconomics Date: 2017-05 Impact factor: 4.981
Authors: Penny J Norsworthy; Jana Vandrovcova; Ellen R A Thomas; Archie Campbell; Shona M Kerr; Jennifer Biggs; Laurence Game; Anne K Soutar; Blair H Smith; Anna F Dominiczak; David J Porteous; Andrew D Morris; Generation Scotland; Timothy J Aitman Journal: BMC Med Genet Date: 2014-06-23 Impact factor: 2.103
Authors: Ruwan A Weerakkody; Jana Vandrovcova; Christina Kanonidou; Michael Mueller; Piyush Gampawar; Yousef Ibrahim; Penny Norsworthy; Jennifer Biggs; Abdulshakur Abdullah; David Ross; Holly A Black; David Ferguson; Nicholas J Cheshire; Hanadi Kazkaz; Rodney Grahame; Neeti Ghali; Anthony Vandersteen; F Michael Pope; Timothy J Aitman Journal: Genet Med Date: 2016-03-24 Impact factor: 8.822