Rapid and accurate denaturating high performance liquid chromatography protocol for the detection of alpha-L-iduronidase mutations causing mucopolysaccharidosis type I
2010
Abstract
We report protocol for a denaturating high performance liquid chromatography (dHPLC), which is used for the rapid and accurate detection of mutations in the IDUA gene. PCR running and dHPLC partial denaturing conditions for mutation detection were established for each PCR amplicon corresponding to 14 IDUA exons and their adjacent intronic/flanking sequences.
Twelve different mutations, 5 nonsense, 4 missense, 1 deletion, and 2 splice site (intron) were screened. All mutations revealed a distinct dHPLC pattern thus enabling their accurate detection.
The results show, that dHPLC method is robust and highly sensitive.