title: [Mutation detection of ADPKD PKD1 gene in Hans by denaturing high-performance liquid chromatography].
abstract: OBJECTIVE
  To develop a screening system for more rapid and sensitive mutation detection of autosomal dominant polycystic kidney disease (ADPKD) gene 1 (PKD1) by using denaturing high-performance liquid chromatography (DHPLC) protocol.
  
  
  METHODS
  Using genomic DNA as templates extracted from blood samples of 19 Han pedigrees with 67 family members, the complete codon areas were amplified by long-range PCR and nested PCR in succession, and then the PCR products were analyzed by DHPLC. The mutations from screened abnormal PCR products were confirmed by DNA sequencing, and then compared with the mutations identified by single strand conformation polymorphism (SSCP) before.
  
  
  RESULTS
  There were 14 mutations found in this study, including 10 missense, 1 insertion, 1 deletion and 2 nonsense mutations. Besides 12 mutations identified before, mutations nt32819G>A and nt37137T>C were the novel mutations found. The mutation detection ratio was 73.7%.
  
  
  CONCLUSIONS
  This developed system via DHPLC can be used as a more effective approach for mutation detection of autosomal dominant polycystic kidney disease PKD1 in Hans.
authors.affiliation: Department of Cell Biology, College of Basic Medical Science, Second Military Medical University, Shanghai, 200433 PR China.
authors.forename: Shu-zhong
authors.initials: SZ
authors.lastname: Zhang
authors.forename: Yu-hong
authors.initials: YH
authors.lastname: Zhang
authors.forename: Dian-yong
authors.initials: DY
authors.lastname: Zhang
authors.forename: Chang-lin
authors.initials: CL
authors.lastname: Mei
mesh_terms: D000328:Adult; D044466:Asian People / Q000235:genetics; D001483:Base Sequence; D002681:China; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D005192:Family Health; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010375:Pedigree; D016891:Polycystic Kidney, Autosomal Dominant / Q000175:diagnosis* / Q000208:ethnology / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D050396:TRPP Cation Channels / Q000235:genetics*
chemical_list: D050396:TRPP Cation Channels