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Background  Mutations in Wilm’s tumor 1 ( WT1 ) gene is one of the commonly reported genetic mutations in children with steroid resistant nephrotic syndrome (SRNS). We report the results of direct sequencing of exons 8 and 9 of  WT1  gene in 100 children with SRNS from a single centre. We standardized and validated High Resolution Melt (HRM) as a rapid and cost effective screening step to identify individuals with normal sequence and distinguish it from those with a potential mutation. Since only mutation positive samples identified by HRM will be further processed for sequencing it will help in reducing the sequencing burden and speed up the screening process.    Methods  One hundred SRNS children were screened for  WT1  mutations in Exon 8 and 9 using Sanger sequencing. HRM assay was standardized and validated by performing analysis for exon 8 and 9 on 3 healthy control and 5 abnormal variants created by site directed mutagenesis and verified by sequencing. To further test the clinical applicability of the assay, we screened additional 91 samples for HRM testing and performed a blinded assessment.    Results    WT1  mutations were not observed in the cohort of children with SRNS. The results of HRM analysis were concordant with the sequencing results.    Conclusion  The  WT1  gene mutations were not observed in the SRNS cohort indicating it has a low prevalence. We propose applying this simple, rapid and cost effective assay using HRM technique as the first step for screening the  WT1  gene hot spot region in a clinical setting.    Electronic supplementary material  The online version of this article (doi:10.1186/s12881-016-0362-7) contains supplementary material, which is available to authorized users.

bmc medical genetics

Screening of WT1 mutations in exon 8 and 9 in children with steroid resistant nephrotic syndrome from a single centre and establishment of a rapid screening assay using high-resolution melting analysis in a clinical setting