Application and evaluation of denaturing HPLC for molecular genetic analysis in tuberous sclerosis

Jones, Alistair C., Sampson, Julian Roy ORCID: https://orcid.org/0000-0002-2902-2348, Hoogendoorn, Bastiaan ORCID: https://orcid.org/0000-0001-9753-169X, Cohen, David and Cheadle, Jeremy Peter ORCID: https://orcid.org/0000-0001-9453-8458 2000. Application and evaluation of denaturing HPLC for molecular genetic analysis in tuberous sclerosis. Human Genetics 106 (6) , pp. 663-668. 10.1007/s004390000316

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Abstract

Tuberous sclerosis (TSC) is an autosomal dom- inant disorder characterised by the development of hamar- tomas in multiple tissues and organs. TSC exhibits locus heterogeneity with genes at 9q34 (TSC1) and 16p13.3 (TSC2) that have 21 and 41 coding exons, respectively. The mutational spectrum at both loci is wide and previous studies have shown that 60%–70% of cases are sporadic and represent new mutations. We have formatted denatur- ing high performance liquid chromatography (DHPLC) for rapid screening of all coding exons of TSC1 and TSC2. DHPLC analysis detected likely disease-causing muta- tions in 103 of 150 unrelated cases (68%), compared with 92/150 (61%) and 87/150 (58%) for single-strand confor- mation polymorphism analysis (SSCP) and conventional heteroduplex analysis (HA), respectively. Capital, con- sumable and labour costs were determined for each exon screening procedure. Estimated costs per patient sample depended on throughput, particularly for DHPLC, where a high proportion of costs are fixed, and were £257, £216 and £242 for DHPLC, SSCP and HA, respectively, as- suming a throughput of 252 samples per year, or £354, £233 and £259, assuming a throughput of 126 samples per year. DHPLC had the advantages of increased sensitivity and reduced labour costs when compared with more tradi- tional approaches to exon screening but, unless expensive DHPLC equipment is being efficiently utilised for a very high proportion of the time available, overall costs are slightly higher.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Uncontrolled Keywords:	Genetics; molecular medicine; gene function; metabolic diseases.
Publisher:	Springer
ISSN:	0340-6717
Last Modified:	25 Oct 2022 09:46
URI:	https://orca.cardiff.ac.uk/id/eprint/59942

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