The Marfan (MFS) and Loeys-Dietz (LDS) syndromes are caused by mutations in the 
fibrillin-1 (FBN1) and Transforming Growth Factor Beta Receptor 1 and 2 (TGFBR1 
and TGFBR2) genes, respectively. With the current conventional mutation 
screening technologies, analysis of this set of genes is time consuming and 
expensive. We have tailored a cost-effective and reliable mutation discovery 
strategy using multiplex PCR followed by Next Generation Sequencing (NGS). In a 
first stage, genomic DNA from five MFS or LDS patient samples with previously 
identified mutations and/or polymorphisms in FBN1 and TGFBR1 and 2 were analyzed 
and revealed all expected variants. In a second stage, we validated the 
technique on 87 samples from MFS patients fulfilling the Ghent criteria. This 
resulted in the identification of 75 FBN1 mutations, of which 67 were unique. 
Subsequent Multiplex Ligation-dependent Probe Amplification (MLPA) analysis of 
the remaining negative samples identified four large deletions/insertions. 
Finally, Sanger sequencing identified a missense mutation in FBN1 exon 1 that 
was not included in the NGS workflow. In total, there was an overall mutation 
identification rate of 92%, which is in agreement with data published 
previously. We conclude that multiplex PCR of all coding exons of FBN1 and 
TGFBR1/2 followed by NGS analysis and MLPA is a robust strategy for time- and 
cost-effective identification of mutations.