title: [The research of combining high resolution melting with multiplex ligation-dependent probe amplification technology in the mutation scanning for PAH gene].
abstract: OBJECTIVE
  To explore the value of combining high resolution melting (HRM) with multiplex ligation-dependent probe amplification (MLPA) for detecting mutations underlying phenylketonuria.
  
  
  METHODS
  HRM was used for detecting small mutations in phenylalanine hydroxylase gene (PAH) of 26 phenylketonuria patients. The results were verified with DNA sequencing. MLPA was used for detecting potential deletions/duplications in the PAH gene. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was performed for additional potential mutations.
  
  
  RESULTS
  A total of 21 mutations were found in 44/52 alleles (84.62%), which included a dupEx4. Among the 21 types of mutation, 19 were reported previously, and the remaining two were novel mutations: c.584_585insA and IVS10+1G>T. In addition, the mutation of R243Q (25%) was the most common type in China.
  
  
  CONCLUSIONS
  The study showed that the combination of HRM and MLPA could increase the detection rate for mutation in PKU. The study has added new information to the human mutation database of PAH and provided a basis for clinical diagnosis and prenatal counseling.
authors.affiliation: The Research Center for Medical Genomics, China Medical University, Shenyang, Liaoning, People's Republic of China.
authors.forename: Chun-yan
authors.initials: CY
authors.lastname: Ji
authors.forename: Li-li
authors.initials: LL
authors.lastname: Sun
authors.forename: Li-hua
authors.initials: LH
authors.lastname: Cao
authors.forename: Yu
authors.initials: Y
authors.lastname: Hu
authors.forename: Hong
authors.initials: H
authors.lastname: Huang
authors.forename: Shu-sen
authors.initials: SS
authors.lastname: Wang
authors.forename: Yang
authors.initials: Y
authors.lastname: Luo
mesh_terms: D019943:Amino Acid Substitution; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D010661:Phenylketonurias / Q000235:genetics*
chemical_list: D010651:Phenylalanine Hydroxylase