Detection of low frequency variants of the NLRP3 gene in “mutation- negative” CAPS patients using massive parallel sequencing

Methods The exons of the NLRP3 gene were amplified via PCR from genomic PBMC DNA. The obtained PCR products were sequenced with an Illumina HiSeq platform. For SNP calling we used the GATK pipeline of the 1000 Genome Project, if the coverage attained 40,000 fold. In order to prove the accuracy of the method, we quantified dilutions of a known heterozygous mutation (T348M) mixed with wildtype DNA. For the correlation between the test results and the phenotype of the patients we developed a survey including symptoms and medical treatment.