Background: The Lesch-Nyhan syndrome is an X-linked recessive inherited disease caused by a complete deficiency of hypoxanthine guanine phosphoribosyl-transferase (HPRT) activity. Many different mutations throughout the HPRT coding region of Lesch-Nyhan patients have been described, including single base substitutions, partial or entire gene deletions, gene insertions or endoduplication of exons. However, study of gene mutation in Chinese patients has rarely been reported in Taiwan.
Methods: Polymerase chain reaction (PCR) and nucleotide sequence analysis were used to identify the location and the nature of the mutation at the HPRT locus in two brothers affected with Lesch-Nyhan syndrome. The HPRT cDNA, amplified from total RNA of patient's peripheral blood by reverse transcription-polymerase chain reaction, was cloned into a pGEM-3Zf(-) vector and then sequenced. Family study involved initial screening using single-strand conformation polymorphism, and further confirmation by direct sequencing of the exon encompassing the mutation.
Results: The mutation identified in these two affected siblings was a single nucleotide substitution, from cytosine to guanine, in exon 3 of the HPRT coding region. This transversion putatively caused a single amino acid substitution from phenylalanine to leucine at codon 74 in the translated protein. This base change was further confirmed by direct sequencing of both the HPRT cDNA fragment and the exon 3 of HPRT gene amplified from genomic DNA. The family study revealed that the patient's mother was a heterozygous carrier, and the mutation seemed to have occurred de novo in a germinal cell from one of the maternal grandparents.
Conclusions: This is the first family study on Chinese patients with Lesch-Nyhan syndrome identified by molecular analysis in Taiwan. The mutation described herein is a novel substitution which occurs in a suggested "hotspot" of mutation (exon 3) of the HPRT gene. The application of molecular analysis of HPRT-gene allows not only DNA diagnosis by directly detecting the mutant alleles, but also prenatal diagnosis and carrier identification within individual families affected by Lesch-Nyhan syndrome.