Sensitive detection of the c-KIT c.1430G>T mutation by mutant-specific polymerase chain reaction in feline mast cell tumours.
Here, we describe the establishment of mutant-specific polymerase chain reaction (PCR) for detection of a c-KIT c.1430G>T mutation in feline mast cell tumours. Several mutations in feline c-KIT have been identified, with the c.1430G>T mutation accounting for a significant portion of feline mast cell tumour mutations. The c.1430G>T mutation in c-KIT exon 9 was detected in 15.7% (11 of 70) of samples by mutant-specific PCR but in only 7.1% (5 of 70) by PCR-restriction fragment length polymorphism (RFLP) in the genomic DNA isolated from 70 formalin-fixed paraffin-embedded sections or cells collected by fine needle aspiration. Mutant-specific PCR showed remarkably higher detection rate than did PCR-RFLP. DNA sequence analysis did not always yield identical results to those of mutant-specific PCR, suggesting heterogeneity of tumour cells. Mutant-specific PCR is a valid and efficient screening tool for detection of the c-KIT c.1430G>T point mutation in feline mast cell tumours compared with PCR-RFLP and sequencing analysis.
Nasunogahara Animal Clinic, Ohtawara, Tochigi, Japan.
M
M
Takanosu
M
M
Sato
Y
Y
Kagawa
D000818:Animals; D002371:Cat Diseases / Q000235:genetics / Q000378:metabolism*; D002415:Cats; D005786:Gene Expression Regulation / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D008113:Liver Neoplasms / Q000378:metabolism / Q000662:veterinary; D034801:Mastocytoma / Q000378:metabolism / Q000662:veterinary*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D012680:Sensitivity and Specificity; D012878:Skin Neoplasms / Q000378:metabolism / Q000662:veterinary; D013160:Splenic Neoplasms / Q000378:metabolism / Q000662:veterinary
D019009:Proto-Oncogene Proteins c-kit