Feasibility and robustness of amplification refractory mutation system (ARMS)-based KRAS testing using clinically available formalin-fixed, paraffin-embedded samples of colorectal cancers.
JAPANESE JOURNAL OF CLINICAL ONCOLOGY(2011)
摘要
KRAS mutation testing is recommended for the discernment of metastatic colorectal cancer patients who are unlikely to benefit from anti-epidermal growth factor receptor antibodies. A recently developed amplification refractory mutation-Scorpion system is becoming a standard method for KRAS mutant detection. The feasibility and robustness of this system using DNA samples from clinically available formalin-fixed, paraffin-embedded specimens were evaluated. Genomic DNA from macro-dissected 110 specimens was applied for the KRAS mutant detection using a commercial amplification refractory mutation-Scorpion system kit. Success rate and mutant detection rate of the test were evaluated. Small intra- and inter-lot deviations of the testing kit and a good concordance among different real-time polymerase chain reaction systems suggested the reliability of the amplification refractory mutation-Scorpion system. Though one-third of the 110 samples that were tested did not contain a sufficient amount of DNA to detect a 1% concentration of mutant alleles, the mutant detection rate was not impaired using tumor DNA concentrated by macro-dissection. Using a higher amount of template DNA, which supposedly contained abundant interfering substances, prevented the detection of the exogenous control amplicons, resulting in a reduced success rate. Adjusting the template amount according to the total DNA concentration might reduce the failure rate. The amplification refractory mutation-Scorpion system with formalin-fixed, paraffin-embedded specimen-derived DNA samples exhibited an acceptable feasibility and robustness suitable for routine clinical practice.
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关键词
colorectal cancer,KRAS mutation,ARMS-Scorpion method,FFPE
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