The frequency of KRAS mutation detection in human colon carcinoma is influenced by the sensitivity of assay methodology: A comparison between direct sequencing and real-time PCR

2010 ◽  
Vol 395 (1) ◽  
pp. 158-162 ◽  
Author(s):  
Takashi Kobunai ◽  
Toshiaki Watanabe ◽  
Yoko Yamamoto ◽  
Kiyoshi Eshima
2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e22100-e22100
Author(s):  
Chi-Kuan Chen

e22100 Background: Identifying K-ras mutation has became a standard procedure in cancer treatment. Colorectal cancer patients with K-ras mutation are likely to poorly respond to cetuximab. Therefore, detecting K-ras gene mutations should be suggested before the selection of personalized treatment in colorectal cancer. To date, general molecular biology techniques contain HRM, PCR-RFLP, TaqMan PCR and CE-IVD-validated Cobas 4800 KRAS (Roche Diagnostics) are used for K-ras mutation detection in molecular diagnosis, but the sensitivity limitation of these method is approximately 1%. Methods: Therefore, we used a new approach, a universal genetic detecting method (FemtoPath), which improves sensitivity of K-ras mutation detection and the limitation of sensitivity is closed to 0.1% of mutation type. Results: We compared the sensitivity between FemtoPath/direct sequencing test and Cobas KRAS real-time PCR. Cobas real-time PCR identifies mutations in 21 (40.38%) of the 52 tumors. Surprisingly, the FemtoPath/direct sequencing test identified mutations in 40 (76.92%) of the 52 tumors. Our data showed that the FemtoPath/direct sequencing test can identify 19 additional mutation samples. In addition, the FemtoPath/direct sequencing test can identify more unknown K-ras mutations adjacent to codon 12 and 13. Conclusions: Ensure the most timely and appropriate therapy for cancer patients is the first priority of clinical application. FemtoPath/direct sequencing test is more sensitive, accurate and inexpensive and needs fewer sample amount than Cobas real-time PCR.


Planta Medica ◽  
2014 ◽  
Vol 80 (16) ◽  
Author(s):  
R Paduch ◽  
M Tomczyk ◽  
A Wiater ◽  
A Dudek ◽  
M Pleszczynska ◽  
...  

1990 ◽  
Vol 2 (10) ◽  
pp. 345-355 ◽  
Author(s):  
Lizabeth Deutsch Murphy ◽  
Eva M. Valverius ◽  
Maria Tsokos ◽  
Lyn A. Mickley ◽  
Neal Rosen ◽  
...  

1990 ◽  
Vol 265 (32) ◽  
pp. 19980-19989
Author(s):  
R V Iozzo ◽  
I Kovalszky ◽  
N Hacobian ◽  
P K Schick ◽  
J S Ellingson ◽  
...  

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