LCHADD Mutation Panel

LCHAD deficiency is a disease that prevents the body from burning certain fats for energy. The age of onset is typically in infancy, and symptoms include feeding problems, lethargy, hypoglycemia, hypotonia, liver problems, abnormalities of the retina, muscle pain, muscle breakdown, and peripheral neuropathy. The disease also has increased heart problems, breathing difficulties, coma, and sudden death risks. The condition can be managed with a strict diet and avoiding periods of fasting. The incidence of LCHAD deficiency is unknown.

Gene analysis for the various targeted mutations is performed by polymerase chain reaction and melting curve analysis to detect the mutant and wild-type forms of the gene. Sequence-specific oligonucleotide probes are labeled with fluorescent dyes, which hybridize to their complementary sequence target in PCR products. The fluorescence resonance energy transfer (FRET) from one fluorophore to another adjacent fluorophore is measurable and is directly proportional to the amount of target DNA generated during PCR. Allele-specific melting curves are generated by slow thermal denaturing of the probe: template hybrid. Melting curves are generated by monitoring fluorescence throughout denaturation, and melting peaks are generated by plotting the inverse derivative of fluorescence versus temperature (−dF/dT).