Calcium-sensing receptor mutations and denaturing high performance liquid chromatography.

[familial hypocalciuric hypercalcemia]

The calcium-sensing receptor (CASR ), a plasma membrane G-protein-coupled receptor , is expressed in parathyroid gland and kidney , and controls systemic calcium homeostasis . Inactivating CASR mutations are associated with familial hypocalciuric hypercalcemia (FHH ) and neonatal severe hyperparathyroidism , and activating mutations cause autosomal dominant hypocalcemia (ADH ). CASR mutation identification plays an important role in the clinical management of mineral metabolism disorders . We describe here a high -throughput method using screening with denaturing high performance liquid chromatography (DHPLC ) to initially interrogate 12 amplicons covering translated exons and exon /intron boundaries , followed by sequencing of any amplicon with a modified melting curve relative to wild type , and direct sequencing of a 13 th amplicon encoding the COOH-terminal tail to distinguish causative mutations from three common missense single nucleotide polymorphisms . A blinded analysis of 32 positive controls representing mutations throughout the CASR sequence , as well as 22 negative controls , yielded a concordance rate of 100 %. We report eight novel and five recurrent FHH mutations , along with six novel and two recurrent ADH mutations . Thus , DHPLC provides a rapid and effective means to screen for CASR mutations .

Diseases
Validation

Diseases presenting "high performance" symptom

alpha-thalassemia

benign recurrent intrahepatic cholestasis

canavan disease

cystinuria

familial hypocalciuric hypercalcemia

neonatal adrenoleukodystrophy

pyruvate dehydrogenase deficiency

You can validate or delete this automatically detected symptom