Detection of an activating c-kit mutation by real-time PCR in patients with anaphylaxis

• Published in: Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Vol. 572; no. 1-2; pp. 1 - 13
• Main Authors: Lawley, Wendy, Hird, Heather, Mallinder, Philip, McKenna, Sue, Hargadon, Beverley, Murray, Alistair, Bradding, Peter
• Format: Journal Article
• Language: English
• Published: Netherlands 02.05.2005
• Subjects: Anaphylaxis - genetics; Aspartic Acid - genetics; Base Sequence; Blotting, Western; Cell Line; Cloning, Molecular; DNA Primers; Humans; Point Mutation; Polymerase Chain Reaction - methods; Proto-Oncogene Proteins c-kit - genetics; Valine - genetics
• ISSN: 0027-5107; 1386-1964; 0027-5107
• DOI: 10.1016/j.mrfmmm.2004.08.015

A well-characterised gain-of-function point mutation within exon 17 of the c-kit proto-oncogene known as Asp816Val is present in patients with mastocytosis. Activation of mast cells through this receptor primes them for IgE-dependent activation, and patients with mastocytosis are at increased risk of anaphylaxis. We hypothesised that the Asp816Val mutation is associated with a history of anaphylaxis in the general population. A mismatch amplification real-time PCR assay was developed and validated to test for the Asp816Val mutation. Subjects were recruited to four subject groups: normal non-atopics, atopics without anaphylaxis, food-induced anaphylactics and non-food anaphylactics. Blood samples collected from forty subjects were tested for the presence of Asp816Val. Thirteen subjects were found to carry the mutation; normals (2/9), atopics (2/10), food anaphylactics (5/11) and non-food anaphylactics (4/10). Statistical analysis of the data determined that there was no significant difference between the numbers of subjects found to carry the Asp816Val mutation in each of the groups although a trend towards an increased occurrence in anaphylactics was observed. In summary, the hypothesis that the presence of the Asp816Val mutation is linked to the occurrence of anaphylaxis was not supported, but interestingly, we have shown for the first time Asp816Val may occur more frequently than previously reported within the general population.