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Massive parallel next-generation sequencing (NGS) technology has dramatically increased the throughput and reduced the cost per nucleotide sequenced compared with traditional Sanger methods, enabling cost-effective sequencing of multiple genes simultaneously.Over the past 3 years, whole-exome sequencing has revealed new genes associated with SRNS in a few cases, expanding the genetic heterogeneity of the disease.Patients developing steroid resistance at a later stage of the disease or with recurrence after kidney transplantation were excluded as we considered that they likely had an immunological cause.Biopsy findings included FSGS, minimal change disease (MCD) or diffuse mesangial sclerosis.Pools of 24 individuals were prepared, hybridized to the custom Nimble Gen Seq Cap EZ Choice Library (Roche Nimble Gen) for 72 h, stringently washed, amplified 17 PCR cycles, and run in a Hi Seq2000 instrument (Illumina Inc.).Data analysis was performed blindly with an in-house developed pipeline previously described.Libraries were prepared with the Tru Seq DNA Sample Preparation Kit (Illumina, Inc.) according to the manufacturer’s instructions.In familial cases, only the proband was analyzed by NGS.
In these cases, renal histology typically shows focal segmental glomerulosclerosis (FSGS).
The study of the relative frequency of mutations in the most commonly altered genes in patients with SRNS and/or FSGS allowed the development of genetic testing algorithms based on age at onset, family history, or renal histology.
However, the genetic heterogeneity and significant phenotypic variability of SRNS make genetic testing using standard Sanger methods costly and time consuming, even if the analysis is restricted to the most frequently mutated genes.
We aimed to develop a glomerular disease gene panel for SRNS/FSGS and to study the influence of mutations in multiple genes on phenotype variability.
A total of 50 Spanish patients with idiopathic SRNS/FSGS were included.