title: [The detection of mutations in VHL gene from a single cell in a patient with von Hippel-Lindau disease].
abstract: OBJECTIVE
  To explore a technology for diagnosing VHL mutations from a single cell and provide experimental evidences for the feasibility of applying technology in detecting genetic mutations from a single cell.
  
  
  METHODS
  After whole genome amplification (WGA) based on multiple displacement amplication (MDA) for a single cell, we did regular PCR following sequencing and detected the genotypes using the real time PCR based on TaqMan probes. We detected VHL mutations by the different terminal fluorescent changing.
  
  
  RESULTS
  The rate of amplification for single cell based on MDA was 90.91%. The rate of contamination was 0. After sequencing, the allele drop out (ADO) rate of heterozygotes was 26.67%(8/30); combined with the different terminal fluorescent changing, the rate of ADO of heterozygotes was 16.67%.
  
  
  CONCLUSIONS
  WGA based on MDA for a single cell followed by regular PCR with sequencing and real time PCR can specially and accurately detect the VHL genotypes of single cells.
authors.affiliation: Department of Urology, Renji Hospital, Medical College of Shanghai Jiaotong University, Shanghai, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Jin
authors.forename: Li-min
authors.initials: LM
authors.lastname: Wang
authors.forename: Ju-fen
authors.initials: JF
authors.lastname: Zheng
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.forename: Yi-ran
authors.initials: YR
authors.lastname: Huang
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis; D005260:Female; D005838:Genotype; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D019836:Preimplantation Diagnosis; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics*; D006623:von Hippel-Lindau Disease / Q000097:blood / Q000235:genetics*
chemical_list: D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human