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Sensitive detection of the c‐ KIT c . 1430G >T mutation by mutant‐specific polymerase chain reaction in feline mast cell tumours
Author(s) -
Takanosu M.,
Sato M.,
Kagawa Y.
Publication year - 2014
Publication title -
veterinary and comparative oncology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 34
eISSN - 1476-5829
pISSN - 1476-5810
DOI - 10.1111/j.1476-5829.2012.00346.x
Subject(s) - mutant , microbiology and biotechnology , polymerase chain reaction , biology , mast cell , mutation , restriction fragment length polymorphism , point mutation , exon , real time polymerase chain reaction , gene , genetics , immunology
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.