High Resolution Melt analysis for mutation screening in PKD1 and PKD2

We aim to prove that high resolution melt analysis (HRM), a recent technique in molecular biology, can facilitate molecular diagnosis of ADPKD. We screened for mutations in PKD1 and PKD2 with HRM in 37 unrelated patients with ADPKD.We identified 440 sequence variants in the 37 patients. One hundred and thirty eight were different. We found 28 pathogenic mutations (25 in PKD1 and 3 in PKD2 ) within 28 different patients, which is a diagnosis rate of 75% consistent with literature mean direct sequencing diagnosis rate. We describe 52 new sequence variants in PKD1 and two in PKD2.HRM analysis is a sensitive and specific method for molecular diagnosis of ADPKD. HRM analysis is also costless and time sparing. Thus, this method is efficient and might be used for mutation pre-screening in ADPKD genes.Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary kidney disorder affecting approximately one in 500 to one in 1000 human live births [1]. It is characterized by focal development and progressive enlargement of renal cysts, leading to end-stage renal disease (ESRD) [2].ADPKD is genetically heterogeneous and involves two genes, PKD1 (MIM 601313, chromosome region 16p13.3) [3] and PKD2 (MIM 173910, 4q21-22) [4]. Mutations in PKD1 account for approximately 85% of ADPKD cases and are associated with a more severe disease than PKD2 . The median age at onset of ESRD is 54.3 [52.7-55.9] years for individuals with mutation in PKD1 compared to 74.0 [67.2-80.8] years in PKD2 [5]. It has been hypothesized that a third gene could be implicated in ADPKD [6] but there is no evidence for this.Genetic analysis of ADPKD is difficult owing to the existence of at least two distinct genes that can cause disease and the lack of an exhaustive list of PKD1 and PKD2 mutations that are associated with it. Genomic features of PKD1 also cause difficulty in identifying sequence variants [7]. The open reading frame of PKD1 is approximately 13 kb split in 46 exons. Exons 1-33 are duplica