OBJECTIVE: To detect pathogenic mutations in Marfan syndrome (MFS) using an Ion 
Torrent Personal Genome Machine (PGM) and to validate the result of targeted 
next-generation semiconductor sequencing for the diagnosis of genetic disorders.
METHODS: Peripheral blood samples were collected from three MFS patients and a 
normal control with informed consent. Genomic DNA was isolated by standard 
method and then subjected to targeted sequencing using an Ion Ampliseq(TM) 
Inherited Disease Panel. Three multiplex PCR reactions were carried out to 
amplify the coding exons of 328 genes including FBN1, TGFBR1 and TGFBR2. DNA 
fragments from different samples were ligated with barcoded sequencing adaptors. 
Template preparation and emulsion PCR, and Ion Sphere Particles enrichment were 
carried out using an Ion One Touch system. The ion sphere particles were 
sequenced on a 318 chip using the PGM platform. Data from the PGM runs were 
processed using an Ion Torrent Suite 3.2 software to generate sequence reads. 
After sequence alignment and extraction of SNPs and indels, all the variants 
were filtered against dbSNP137. DNA sequences were visualized with an Integrated 
Genomics Viewer. The most likely disease-causing variants were analyzed by 
Sanger sequencing.
RESULTS: The PGM sequencing has yielded an output of 855.80 Mb, with a > 100 × 
median sequencing depth and a coverage of > 98% for the targeted regions in all 
the four samples. After data analysis and database filtering, one known missense 
mutation (p.E1811K) and two novel premature termination mutations (p.E2264X and 
p.L871FfsX23) in the FBN1 gene were identified in the three MFS patients. All 
mutations were verified by conventional Sanger sequencing.
CONCLUSION: Pathogenic FBN1 mutations have been identified in all patients with 
MFS, indicating that the targeted next-generation sequencing on the PGM 
sequencers can be applied for accurate and high-throughput testing of genetic 
disorders.