Detection of tumor-derived DNA in exfoliated cells of urine sediments with a novel quantitative PCR method based on allele-specific PCR.
We have established a novel quantitative method based on the allele-specific PCR, which uses the linearly amplified fragment of the PCR products as the internal control. The improved characteristics of the procedure are the high sensitivity for quantitation of the mutant alleles at ratios of up to 1:10000 and the reduced necessity of the optimization of the PCR conditions for each mutation. Using this modified allele-specific PCR, we could quantify the tumor alleles in the urine sediments of three patients with urothelial cancers that harbored different p53 gene mutations. This method can be applied to other genetic targets that have other types of alterations, such as deletions or insertions.
Yamada
H
H
Department of Urology, Faculty of Medicine, Kyoto University, Japan.
Kakehi
Y
Y
Okuno
H
H
Yoshida
O
O
D000483:Alleles; D004273:DNA, Neoplasm / Q000652:urine*; D016158:Genes, p53; D006801:Humans; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000652:urine; D014571:Urologic Neoplasms / Q000235:genetics / Q000652:urine*
D004273:DNA, Neoplasm