RESUMO
The knowledge of at least 21 different mutations and several polymorphisms in the coproporphyrinogen oxidase (CPO) gene demonstrates that the molecular basis of hereditary coproporphyria is heterogeneous. We developed a DGGE-based assay for the analysis of exons 2 to 7, including 14-96 nucleotides of the flanking intronic sequences of the CPO gene. To render it suitable for the clinical diagnostic laboratory, we designed the assay to allow use of identical PCR conditions and the same DGGE gel for analyses of all the regions. Using this assay, and subsequent sequencing of gene regions containing interallelic variations, two novel mutations in the CPO gene were identified: a missense mutation (607G-->A), leading to the substitution of an alanine with a threonine, and a nonsense mutation (1281G-->A), giving rise to a stop codon 28 codons upstream to the wild-type stop codon.
Assuntos
Coproporfirinogênio Oxidase/genética , DNA/genética , Eletroforese em Gel de Poliacrilamida/métodos , Mutação , Porfirias Hepáticas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Coproporfirinogênio Oxidase/química , Primers do DNA , Dinamarca , Feminino , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Porfirias Hepáticas/enzimologia , Porfirias Hepáticas/etnologia , Homologia de Sequência de AminoácidosRESUMO
Simple screening tests, urinary porphobilinogen (PBG) for acute intermittent porphyria (AIP) and fecal coproporphyrin for hereditary coproporphyria (HCP), were performed in a family study of AIP and HCP. Urinary PBG was positive in 93 of 211 members of 10 AIP families, but was negative in 568 of 572 controls. Fecal coproporphyrin was positive in 54 of 108 members of 10 HCP families, but was negative in 188 controls. A dominant inheritance was assumed by a chi-square test and Weinberg segregation ratio. Worsening factors around puberty were suggested by the onset age and cumulative percentage of genetically loaded cases. Sex-related expression of symptoms was also inferred by a higher incidence of both porphyrias in females than in males. Fitness and penetrance of both porphyrias were good. An l-triiodothyronine loading test was the most useful for the detection of masked carriers of AIP. In conclusion, AIP and HCP in Japan show a dominant inheritance with sex-related metabolic and clinical manifestations.