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Detection of K‐ ras mutation in sputum by mutant‐allele‐specific amplification (MASA)
Author(s) -
Takeda Satoshi,
Ichii Shigetoshi,
Nakamura Yusuke
Publication year - 1993
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380020209
Subject(s) - biology , mutant , allele , sputum , mutation , microbiology and biotechnology , gene , lung cancer , exon , oligonucleotide , polymerase chain reaction , genetics , cancer research , pathology , tuberculosis , medicine
From 10 to 30% of lung carcinomas examined to date contain mutant K‐ras genes. We report here that the mutant‐allele‐specific amplification (MASA) method may be useful for detection of the K‐ ras mutations in cells obtained from the sputum of patients with lung cancer. The PCR product from one of five patients revealed an alteration when mixed oligonucleotides representing variants of the second letter at codon 12 of this gene were used as 5′ primers, and further experiments showed a mutation of GGT (Gly) to GAT (Asp)at codon 12. The MASA system could also be applied to an examination of metastatic lung carcinomas, particularly from adenocarcinomas in colon and pancreas in which frequent K‐ ras mutations are detected, and to mass‐screening for colorectal tumors using DNA isolated from feces as template. © 1993 Wiley‐Liss, Inc.