High-resolution melting analysis, a simple and effective method for reliable mutation scanning and frequency studies in the ACADVL gene

Rikke Katrine Jentoft Olsen, Steven F. Dobrowolski, Margrethe Kjeldsen, David Hougaard, Henrik Simonsen, Niels Gregersen, Brage Storstein Andresen

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Expanded newborn screening uses tandem mass spectrometry (MS/MS) to identify patients affected with fatty acid oxidation defects by the presence of pathological acylcarnitine esters. A caveat to MS/MS assessment is that cut-off values for disease-specific acylcarnitines does not always clearly discriminate affected patients from carriers and healthy individuals. Diagnostic evaluation of screening-positive samples is required to confirm a metabolic deficiency. With MS/MS newborn screening becoming established in a growing number of countries, streamlined means for time-and-effective follow-on diagnostic evaluation is essential. Moreover, studies to evaluate the diagnostic accuracy of MS/MS newborn screening are needed for determination and adjustment of precise cut-off values for the disease-specific acylcarnitines. In the current study, we use the fatty acid oxidation disorder very-long-chain acyl-CoA dehydrogenase deficiency (VLCADD), the second most common fatty acid oxidation disorder detected by expanded newborn screening, to demonstrate accurate and fast diagnostic evaluation of the ACADVL gene utilizing DNA extracted from the newborn screening dried bloodspot and high resolution melt (HRM) profiling. We also demonstrate that HRM is a very effective means to determine carrier frequency of prevalent ACADVL mutations in the general population. Based on estimates of the expected disease incidence, we discuss the diagnostic accuracy of MS/MS-based newborn screening to identify VLCADD in Denmark.

TidsskriftJournal of Inherited Metabolic Disease
Udgave nummer3
Sider (fra-til)247-260
StatusUdgivet - jun. 2010
Udgivet eksterntJa

Bibliografisk note

Funding Information:
Acknowledgments This work was supported by grants from The Danish Medical Research Council, the March of Dimes Foundation (grant no.1-FY-2003-688), The Aase and Ejnar Danielsen Foundation, Læge Sofus Carl Emil Friis og Hustru Olga Friis’ legat, Vanførefonden and Oda and Hans Svenningsen Foundation.

Funding Information:
References to electronic databases: OMIM disorder: http://www.ncbi.nlm.nih.gov/omim Enzyme Commission (EC) number: http://www.chem.qmul.ac.uk/ iubmb/enzyme/ GenBank: http://www.ncbi.nlm.nih.gov/Genbank/ HGVS nomenclatures: http://www.hgvs.org/mutnomen/ The DNA mfold server programme (http://mfold.bioinfo.rpi.edu/cgi-bin/rna-form1.cgi/) Competing interest: Steven F. Dobrowolski is former employee at Idaho Technology. Rikke KJ Olsen presented preliminary data from the article at the 55th Annual Meeting, The American Society of Human Genetics. Travel expenses were supported financially by Idaho Technology. The content of the article has not been influenced by this connection to Idaho Technology nor by any other competing financial or non-financial interests.


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