Use of the single-strand conformational polymorphism method to detect recurrent and novel mutations in the low-density lipoprotein receptor gene in patients with familial hypercholesterolaemia: detection of a novel mutation Asp200-->Gly.

The single-strand conformational polymorphism (SSCP) method was used to look for mutations in the 3' half of exon 4 of the low-density lipoprotein receptor gene in patients with familial hypercholesterolaemia (FH). One set of conditions were found which allowed the detection of four of the mutations that have previously been reported in this part of the gene and detected in patients in the United Kingdom: the 3-bp deletion (del Gly197) the 2-bp deletion (STOP 216), the Asp206-->Glu mutation and the Cys210-->STOP. The method was used to screen 50 patients with definite or probable FH from London. Two were identified who were carriers of the 3-bp deletion of Gly197, one who was a carrier of the Asp206-->Glu mutation and one who was a carrier of a novel mutation that alters Asp200-->Gly. This mutation creates a cutting site for the restriction enzyme MspI. In a further sample of 200 patients from London with FH one additional apparently unrelated individual was detected who was a carrier of this defect. Thus in the sample of 50 patients, four (8%) had a mutation in this part of exon 4 that could be readily detected using the SSCP method, suggesting that this approach will be useful for rapid screening for mutations in patients with FH.