Identification of novel mutations in Chinese Hans with autosomal dominant polycystic kidney disease

Abstract

BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is the most common inherited renal disease with an incidence of 1 in 400 to 1000. The disease is genetically heterogeneous, with two genes identified: PKD1 (16p13.3) and PKD2 (4q21). Molecular diagnosis of the disease in at-risk individuals is complicated due to the structural complexity of PKD1 gene and the high diversity of the mutations. This study is the first systematic ADPKD mutation analysis of both PKD1 and PKD2 genes in Chinese patients using denaturing high-performance liquid chromatography (DHPLC).MethodsBoth PKD1 and PKD2 genes were mutation screened in each proband from 65 families using DHPLC followed by DNA sequencing. Novel variations found in the probands were checked in their family members available and 100 unrelated normal controls. Then the pathogenic potential of the variations of unknown significance was examined by evolutionary comparison, effects of amino acid substitutions on protein structure, and effects of splice site alterations using online mutation prediction resources.ResultsA total of 92 variations were identified, including 27 reported previously. Definitely pathogenic mutations (ten frameshift, ten nonsense, two splicing defects and one duplication) were identified in 28 families, and probably pathogenic mutations were found in an additional six families, giving a total detection level of 52.3% (34/65). About 69% (20/29) of the mutations are first reported with a recurrent mutation rate of 31%.ConclusionsMutation study of PKD1 and PKD2 genes in Chinese Hans with ADPKD may contribute to a better understanding of the genetic diversity between different ethnic groups and enrich the mutation database. Besides, evaluating the pathogenic potential of novel variations should also facilitate the clinical diagnosis and genetic counseling of the disease.

Comprehensive PKD1 and PKD2 Mutation Analysis in Prenatal Autosomal Dominant Polycystic Kidney Disease

“…They are shown in Supplemental Table 2, with the scores established to evaluate their putative pathogenicity by different programs (see Methods). Six were previously reported, 10,[12][13][14][15] and three were present in our in-house database: p.Thr2710N and p.Arg3183Gln were associated with another PKD1 or PKD2 mutation in patients diagnosed in the fourth decade, and the p.R3277C mutation was identified at the homozygous state in a woman transplanted at 50 years. No additional PKD1 or PKD2 variations were identified in the other patients.…”

Analysis of gene mutations in PKD1/PKD2 by multiplex ligation-dependent probe amplification: some new findings

“…9 Li et al 10 found the mutation c.3623-3624 insGTGT in exon 15 of PKD1 using combined denatured high-performance liquid chromatography (DHPLC) and gene sequencing. Yu et al 11 detected a series of mutations in 65 autocephalous lineages. If all the exons of PKD1 and PKD2 were to be screened for mutations using amplification and sequencing, it would not only be difficult to design the primers and perform elaborate PCR methodologies, but also a large number of samples would be required.…”

A New PKD1 Mutation Discovered in a Chinese Family with Autosomal Polycystic Kidney Disease

“…The presence of these highly homologous pseudogenes has made genetic analysis of PKD1 challenging, thereby discrimination of these pseudogenes from PKD1 requires locus-specific amplification methods. Although some researchers performed denaturing high-performance liquid chromatography (DHPLC) and polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) to screen mutations in ADPKD for research and clinical purposes [17,18], direct sequencing is considered to be a better choice [18]. Recently, Tan et al proposed a novel LR PCR Sequencing Method [14].…”