PMID- 31984575
OWN - NLM
STAT- MEDLINE
DCOM- 20210708
LR  - 20210708
IS  - 1521-2254 (Electronic)
IS  - 1099-498X (Linking)
VI  - 22
IP  - 5
DP  - 2020 May
TI  - Implementation of high-resolution melting analysis of the porcupine (PORCN) gene 
      for molecular diagnosis of focal dermal hypoplasia: Identification of a novel 
      mutation.
PG  - e3165
LID - 10.1002/jgm.3165 [doi]
AB  - BACKGROUND: Focal dermal hypoplasia (FDH) is rare X-linked dominant disease 
      characterized by atrophy and linear pigmentation of the skin, split hand/foot 
      deformities and ocular anomalies. FDH is caused by mutations of the Porcupine 
      (PORCN) gene, which encodes an enzyme that catalyzes the palmitoylation of Wnt 
      ligands required for their secretion. High resolution melting analysis (HRM) is a 
      technique that allows rapid, labor-efficient, low-cost detection of genomic 
      variants. In the present study, we report the successful implementation of HRM in 
      the molecular diagnosis of FDH. METHODS: Polymerase chain reaction and HRM assays 
      were designed and optimized for each of the coding exons of the PORCN gene, 
      processing genomic DNA samples form a non-affected control and a patient 
      complying with the FDH diagnostic criteria. The causal mutation was characterized 
      by Sanger sequencing from an amplicon showing a HRM trace suggesting heterozygous 
      variation and was validated using an amplification-refractory mutation system 
      (ARMS) assay. RESULTS: The melting profiles suggested the presence of a variant 
      in the patient within exon 1. Sanger sequencing revealed a previously unknown C 
      to T transition replacing a glutamine codon for a premature stop codon at 
      position 28, which was validated using ARMS. CONCLUSIONS: Next-generation 
      sequencing facilitates the molecular diagnosis of monogenic disorders; however, 
      its cost-benefit ratio is not optimal when a single, small or medium size causal 
      gene is already identified and the clinical diagnostic presumption is strong. 
      Under those conditions, as it is the case for FDH, HRM represents a cost- and 
      labor-effective approach.
CI  - (c) 2020 John Wiley & Sons, Ltd.
FAU - Martinez-Saucedo, Mirna
AU  - Martinez-Saucedo M
AD  - Laboratory of Research in Genomics, Genetics and Bioinformatics, Hospital 
      Infantil de Mexico Federico Gomez, Mexico City, Mexico.
FAU - Ornelas-Fuentes, Carolina
AU  - Ornelas-Fuentes C
AD  - Department of Medical Genetics and Experimental Teratogenesis, Hospital Infantil 
      de Mexico Federico Gomez, Mexico City, Mexico.
FAU - Dedden, Mark
AU  - Dedden M
AD  - Laboratory of Research in Genomics, Genetics and Bioinformatics, Hospital 
      Infantil de Mexico Federico Gomez, Mexico City, Mexico.
FAU - Sanchez-Urbina, Rocio
AU  - Sanchez-Urbina R
AD  - Laboratory of Developmental Biology and Experimental Teratogenesis, Hospital 
      Infantil de Mexico Federico Gomez,, Mexico City, Mexico.
FAU - Diaz-Garcia, Hector
AU  - Diaz-Garcia H
AD  - Laboratory of Developmental Biology and Experimental Teratogenesis, Hospital 
      Infantil de Mexico Federico Gomez,, Mexico City, Mexico.
FAU - Aquino-Jarquin, Guillermo
AU  - Aquino-Jarquin G
AUID- ORCID: 0000-0003-4762-6695
AD  - Laboratory of Research in Genomics, Genetics and Bioinformatics, Hospital 
      Infantil de Mexico Federico Gomez, Mexico City, Mexico.
FAU - Moreno-Salgado, Rodrigo
AU  - Moreno-Salgado R
AD  - Department of Medical Genetics and Experimental Teratogenesis, Hospital Infantil 
      de Mexico Federico Gomez, Mexico City, Mexico.
FAU - Granados-Riveron, Javier T
AU  - Granados-Riveron JT
AUID- ORCID: 0000-0001-5485-654X
AD  - Laboratory of Research in Genomics, Genetics and Bioinformatics, Hospital 
      Infantil de Mexico Federico Gomez, Mexico City, Mexico.
LA  - eng
PT  - Case Reports
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200216
PL  - England
TA  - J Gene Med
JT  - The journal of gene medicine
JID - 9815764
RN  - 0 (Codon, Nonsense)
RN  - 0 (Membrane Proteins)
RN  - EC 2.3.- (Acyltransferases)
RN  - EC 2.3.1.- (PORCN protein, human)
SB  - IM
MH  - Acyltransferases/*genetics
MH  - Amino Acid Sequence
MH  - Codon, Nonsense
MH  - Exons/*genetics
MH  - Female
MH  - Focal Dermal Hypoplasia/*diagnosis/*genetics/physiopathology
MH  - Heterozygote
MH  - Humans
MH  - Infant
MH  - Membrane Proteins/*genetics
MH  - Mutation
MH  - Nucleic Acid Denaturation/*genetics
MH  - Phylogeny
MH  - Polymerase Chain Reaction/methods
MH  - Sequence Alignment
OTO - NOTNLM
OT  - PORCN
OT  - focal dermal hypoplasia
OT  - high-resolution melting analysis
EDAT- 2020/01/28 06:00
MHDA- 2021/07/09 06:00
CRDT- 2020/01/28 06:00
PHST- 2019/11/25 00:00 [received]
PHST- 2020/01/18 00:00 [revised]
PHST- 2020/01/19 00:00 [accepted]
PHST- 2020/01/28 06:00 [pubmed]
PHST- 2021/07/09 06:00 [medline]
PHST- 2020/01/28 06:00 [entrez]
AID - 10.1002/jgm.3165 [doi]
PST - ppublish
SO  - J Gene Med. 2020 May;22(5):e3165. doi: 10.1002/jgm.3165. Epub 2020 Feb 16.