Comparison of two different screening methods for the KRAS mutation in colorectal cancer.
BACKGROUND
To compare high resolution melting (HRM) and primer extension (PE) for mutation screening of KRAS codons 12 and 13.


METHODS
DNA samples were isolated from 60 colorectal cancer (CRC) patients. HRM and primer extension analyses were used for mutation screening of KRAS codons 12 and 13.


RESULTS
Both methods can detect KRAS mutations in a DNA mixture containing as little as 5% mutant DNA. The concordant rate between the two methods was 100%. HRM analysis is able to distinguish mutant from wild type, however, it is unable to detect the actual base change. PE analysis needs more procedures, is time-consuming and slightly more expensive but it is able to detect the precise mutation.


CONCLUSIONS
This study showed that HRM is a reliable screening method to identify mutations in the KRAS gene, and the PE can be used as a diagnostic method to accurately pinpoint the nature of mutations.
Er
Tze-Kiong
TK
Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Chang
Ya-Sian
YS
Yeh
Kun-Tu
KT
Chang
Tien-Jye
TJ
Chang
Jan-Gowth
JG
D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D001483:Base Sequence; D045744:Cell Line, Tumor; D003062:Codon / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D006147:Guanine; D006801:Humans; D008403:Mass Screening / Q000379:methods; D009154:Mutation; D009691:Nucleic Acid Denaturation / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D017384:Sequence Deletion; D013941:Thymine; D018631:ras Proteins / Q000235:genetics*
D003062:Codon; D017931:DNA Primers; D004273:DNA, Neoplasm; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D006147:Guanine; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins; D013941:Thymine