title: Molecular genetic analysis of PKHD1 by next-generation sequencing in Czech families with autosomal recessive polycystic kidney disease.
abstract: BACKGROUND
  Autosomal recessive polycystic kidney disease (ARPKD) is an early-onset form of polycystic kidney disease that often leads to devastating outcomes for patients. ARPKD is caused by mutations in the PKHD1 gene, an extensive gene that encodes for the ciliary protein fibrocystin/polyductin. Next-generation sequencing is presently the best option for molecular diagnosis of ARPKD. Our aim was to set up the first study of ARPKD patients from the Czech Republic, to determine the composition of their mutations and genotype-phenotype correlations, along with establishment of next-generation sequencing of the PKHD1 gene that could be used for the diagnosis of ARPKD patients.
  
  
  METHODS
  Mutational analysis of the PKHD1 gene was performed in 24 families using the amplicon-based next-generation sequencing (NGS) technique. In patients without 2 causal mutations identified by NGS, subsequent MLPA analysis of the PKHD1 gene was carried out.
  
  
  RESULTS
  Two underlying mutations were detected in 54% of families (n = 13), one mutation in 13% of families (n = 3), and in 33% of families (n = 8) no mutation could be detected. Overall, seventeen different mutations (5 novel) were detected, including deletion of one exon. The detection rate in our study reached 60% in the entire cohort of patients; but 90% in the group of patients who fulfilled all clinical criteria of ARPKD, and 42% in the group of patients with unknown kidney pathology. The most frequent mutation was T36M, accounting for nearly 21% of all identified mutations.
  
  
  CONCLUSIONS
  Next-generation sequencing of the PKHD1 gene is a very useful method of molecular diagnosis in patients with a full clinical picture of ARPKD, and it has a high detection rate. Furthermore, its relatively low costs and rapidity allow the molecular genetic analysis of patients without the full clinical criteria of ARPKD, who might also have mutations in the PKHD1 gene.
authors.affiliation: Institute of Biology and Medical Genetics of the First Faculty of Medicine, General University Hospital in Prague, Prague, Czech. lena.obeidova@gmail.com.
authors.forename: Lena
authors.initials: L
authors.lastname: Obeidova
authors.affiliation: Department of Paediatrics, 2nd Faculty of Medicine, Charles University in Prague and Motol University Hospital, Prague, Czech. tomas.seeman@lfmotol.cuni.cz.
authors.forename: Tomas
authors.initials: T
authors.lastname: Seeman
authors.affiliation: Institute of Biology and Medical Genetics of the First Faculty of Medicine, General University Hospital in Prague, Prague, Czech. velisakova@post.cz.
authors.forename: Veronika
authors.initials: V
authors.lastname: Elisakova
authors.affiliation: Department of Nephrology, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Prague, Czech. JReiterova@seznam.cz.
authors.forename: Jana
authors.initials: J
authors.lastname: Reiterova
authors.affiliation: Department of Biology and Medical Genetics, 2nd Faculty of Medicine, Charles University in Prague and Motol University Hospital, Prague, Czech. Alena.puchmajerova@lfmotol.cuni.cz.
authors.forename: Alena
authors.initials: A
authors.lastname: Puchmajerova
authors.affiliation: Institute of Biology and Medical Genetics of the First Faculty of Medicine, General University Hospital in Prague, Prague, Czech. jstek@lf1.cuni.cz.
authors.forename: Jitka
authors.initials: J
authors.lastname: Stekrova
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D018153:Czech Republic; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons / Q000235:genetics; D005192:Family Health; D005787:Gene Frequency; D005808:Genes, Recessive; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007223:Infant; D007438:Introns / Q000235:genetics; D009154:Mutation; D017044:Polycystic Kidney, Autosomal Recessive / Q000175:diagnosis / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011956:Receptors, Cell Surface / Q000235:genetics*
chemical_list: C451750:PKHD1 protein, human; D011956:Receptors, Cell Surface