Detection of colorectal cancer K-ras mutations using a simplified oligonucleotide ligation assay
Detection of colorectal cancer K-ras mutations using a simplified oligonucleotide ligation assay
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DOI：
10.1016/s0022-1759(97)00078-1
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发表时间：
1997-08-07
影响因子：
2.2
通讯作者：
Triscott, MX
中科院分区：
文献类型：
--
作者：
Rothschild, CB;Brewer, CS;Triscott, MX
已有研究表明，K-ras基因第12、13密码子的某些突变与结直肠腺瘤的癌变有关。这项研究的目的是建立一种快速、比色的方法来检测通常与结直肠癌相关的K-ras点突变。从结直肠癌DNA中扩增出K-ras外显子1，并结合免疫学和比色法检测到K-ras激活突变。使用OLA和针对单个K-ms突变的寡核苷酸，发现17个结直肠腺瘤/癌中有6个存在K-ras突变。该方法可以检测到低至10%的突变等位基因。开发了一种简化的OLA，旨在测试任何K-RNS激活突变的存在(+)或不存在(-)，通过使用比色显影的试纸方法进一步简化了检测。如果需要，可以通过使用最近描述的EDNA-ELCA检测系统来提高检测灵敏度。简化的(+/-)突变Ola结合试纸或EDNA-ELCA检测系统提供了一种快速、灵敏的K-ras点突变检测方法，适用于结直肠癌的临床评估。(C)1997年爱思唯尔科学公司。
It has been suggested that some mutations in codons 12 and 13 of the K-ras gene are associated with the progression of colorectal adenomas to carcinomas. The aim of this study was to develop a rapid, colorimetric assay for K-ras point mutations commonly associated with colorectal cancer. K-ras exon 1 was amplified from colorectal tumor DNA and K-ras activating mutations detected using an oligonucleotide ligation assay (OLA) in combination with immunological and colorimetric detection. Using the OLA with oligonucleotides specific to individual K-ms mutations, 6 (of 17 total colorectal adenomas/carcinomas) were found to have K-ras mutations. The assay could detect as little as 10% mutant allele. A simplified OLA designed to test for either the presence (+) or absence (-) of any of the K-rns activating mutations was developed, The assay was further streamlined by use of a dipstick methodology for colorimetric development. If required, assay sensitivity can be increased by the use of the recently described EDNA-ELCA detection system. The simplified (+/-) mutation OLA in combination with a dipstick or EDNA-ELCA detection system provides a rapid, sensitive assay for K-ras point mutations suitable for use as part of the clinical assessment of colorectal cancer. (C) 1997 Elsevier Science B.V.