【摘 要】
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Circulating tumor cells in blood from metastatic breast and lung cancer patients have been reported as a surrogate marker for tumor response and shorter sur
【机 构】
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St.Director, Genomic Assays R&D Molecular Biology Systems Division, Life Technologies Corporation US
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Circulating tumor cells in blood from metastatic breast and lung cancer patients have been reported as a surrogate marker for tumor response and shorter survival.Detection of these rare tumor cells in blood is important for early cancer detection.Here we report a new competitive allele-specific TaqMan-based PCR (castPCR) method for rare mutation detection.CastPCR combines allele-specific TaqMan qPCR with allele-specific minor grove binder (MGB) as blockers to effectively suppress non-specific amplification from the wild type allele.We have successfully designed and validated castPCR assays for more than 80 cancer-related SNPs of RAS, EGFR, KIT, pTEN, p53 and others.Results demonstrate that castPCR not only maintains wide dynamic range, high sensitivity, and reproducibility of TaqMan assays but also improve assay specificity of detecting rare mutant alleles for up to 1 in 100,000,000.Our data suggest castPCR is capable of detecting circulating tumor cells in blood.
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