Feasibility and Robustness of Amplification Refractory Mutation System (ARMS)-based KRAS Testing Using Clinically Available Formalin-fixed, Paraffin-embedded Samples of Colorectal Cancers

作者：Ogasawara Naomi; Bando Hideaki; Kawamoto Yasuyuki; Yoshino Takayuki; Tsuchihara Katsuya; Ohtsu Atsushi; Esumi Hiroyasu^*

来源：Japanese Journal of Clinical Oncology, 2011, 41(1): 52-56.

DOI：10.1093/jjco/hyq151

• colorectal cancer
• KRAS mutation
• ARMS-Scorpion method
• FFPE

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.

• 出版日期2011-1