The use of COLD-PCR, DHPLC and GeneScanning for the highly sensitive detection of c-KIT somatic mutations in canine mast cell tumours

摘要: The conventional polymerase chain reaction (PCR)/sequencing methods may be poorly suited for the detection of somatic mutations in canine mast cell tumour (MCT) samples owing to limited sensitivity. This study was aimed at establishing novel and more sensitive methods, assessing their limit comparing sensitivity with methods.Two different ‘driver’ c-KIT, together wild-type counterparts, were cloned plasmids prepare standard known concentrations mutated alleles a background alleles; standards assayed using either or novel, highly technique. Conventional PCR/sequencing showed 50–20%. Conversely, all obtained higher sensitivities allowed reaching as low 2.5–1.2% DNA.The demonstrates that early could likely have underestimated prevalence KIT MCTs, therefore affecting assessment relevance prognosis tyrosine kinase inhibitor (TKI) treatment effectiveness.