Genetic analysis
DNA was isolated from peripheral blood samples using the MagNa Pure
Compact Nucleic Acid Isolation Kit I (Roche) according to the
manufacturer’s instructions. A next generation sequencing (NGS) panel
including genes related to CMC (STAT1, TRAF3IP2, CARD9, IL17F,
IL17RA, and IL17RC ), was performed using an AmpliSeq strategy on
the Ion Torrent PGM platform. Primary sequence data analysis was
performed with Torrent server 5.1 software. Single-nucleotide
polymorphisms were denoted as proposed by the Torrent Variant Caller
v5.12.0.40 and analyzed by Ion Reporter Sofware v5.12. Identified
specific mutations were validated by Sanger sequencing using specific
primers amplifying exon 6 of TRAF3IP2 gene and exon 17 ofSTAT1 .