Noninvasive detection through REMS-PCR technique of K-ras mutations in stool DNA of patients with colorectal cancer.
Noninvasive detection through REMS-PCR technique of K-ras mutations in stool DNA of patients with colorectal cancer.
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发表时间：
2007-03
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通讯作者：
F. Mixich;M. Ioana;F. Voinea;A. Saftoiu;T. Ciurea
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作者：
F. Mixich;M. Ioana;F. Voinea;A. Saftoiu;T. Ciurea
背景和目的脱落到粪便中的肿瘤脱落细胞是结肠恶性肿瘤分子筛查和早期检测的有吸引力的目标。许多研究表明，检测激活的 ras 可能具有诊断或预后的重要性。本研究的目的是确定粪便中 K-ras 突变测定的无创筛查测试在诊断实验室中的适用性及其对结直肠癌的常规预后价值。方法 对 28 名仅通过组织病理学结果诊断为原发性结直肠癌的患者进行息肉切除和结直肠活检后获得的配对粪便和组织标本，通过限制性内切酶介导的选择性 (REMS)-PCR 前瞻性研究 K-ras 密码子 12 突变。结果 28 份组织样本（100%）和 26 份粪便样本（92.8%）获得 DNA。在 14 份 (50.0%) 配对粪便和组织样本中发现 K-ras 密码子 12 突变。突变检测可以在野生型序列的 1000 倍过量的情况下进行。这些结果对于结肠恶性肿瘤患者的基因筛查方案的设计、预后的确定、早期检测和治疗可能具有重要意义。结论 K-ras 测定结直肠癌患者粪便来源 DNA 的敏感性和特异性支持开展大规模试验以验证其作为筛查试验的用途。 REMS-PCR 不是劳动密集型的，而是一种灵敏、快速且稳健的点突变检测方法，由我们在常规诊断实验室中引入。
BACKGROUND AND AIMS Tumor exfoliated cells that shed into stool are attractive targets for molecular screening and early detection of colon malignancies. Many studies have suggested that the detection of activated ras may have diagnostic or prognostic importance. The aim of this study was to establish the suitability for use in diagnostic laboratories of the noninvasive screening test of K-ras mutation determination in the stool and its routine prognostic value in colorectal cancer. METHODS Paired stool and tissue specimens obtained after polypectomy and colorectal biopsy from 28 patients diagnosed solely by histopathological findings with primary colorectal carcinoma, were prospectively studied for K-ras codon 12 mutations by restriction endonuclease-mediated selective (REMS)-PCR. RESULTS DNA was obtained in 28 of tissue samples (100%) and 26 of stool samples (92.8%). K-ras codon 12 mutation was seen in 14 (50.0%) paired stool and tissue samples. Mutation detection was possible in 1000-fold excess of wild-type sequence. These results may be important in the design of genetic screening programs, determination of prognosis, early detection and treatment for patients with colon malignancy. CONCLUSIONS The sensitivity and specificity of K-ras determination on stool-derived DNA in patients with colorectal carcinoma, support the opportunity of a large-scale trial to validate its use as a screening test. REMS- PCR is not labor intensive, but a sensitive, rapid, and robust assay for the detection of point mutations, and was introduced by us in a routine diagnostic laboratory.