Detecting colorectal cancer in stool with the use of multiple genetic targets
Detecting colorectal cancer in stool with the use of multiple genetic targets
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DOI：
10.1093/jnci/93.11.858
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发表时间：
2001-06-06
期刊：
影响因子：
--
通讯作者：
Jen, J
中科院分区：
文献类型：
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作者：
Dong, SM;Traverso, G;Jen, J
背景：结直肠癌细胞脱落到粪便中，为使用无创方法早期检测疾病提供了一种潜在的手段。我们的目标是开发可靠的、特异的分子基因检测方法来检测粪便样本中的结直肠癌。方法：从51例结直肠癌患者的配对粪便和原发肿瘤样本中分离粪便DNA。三个基因靶点- tp53， BAT26和k - ras -被用于在配对肿瘤的分子分析之前或不考虑的情况下检测粪便中的肿瘤相关突变。TP53基因突变检测与错配连接试验，检测9个常见的p53基因突变。采用改进的固相微序列法检测BAT26基因座的缺失。采用基于数字聚合酶链反应的方法检测K-RAS密码子12和13的突变。结果：在30例患者的肿瘤DNA中检测到TP53基因突变，所有患者的粪便中都有相同的TP53突变。来自三名患者的肿瘤在BAT26位点含有非遗传性缺失，并且在这些患者的粪便标本中发现了相同的改变。50个被检测的肿瘤中有19个有KRAS突变；在8名患者的配对粪便DNA样本中检测到相同的突变。没有一例在粪便中发现的突变在原发肿瘤中也存在。因此，这三种遗传标记共同检测出51例（95%置信区间[CI] = 56%至83%）结直肠癌患者中的36例（71%）和39例（95% CI = 79%至98%）肿瘤发生改变的患者中的36例（92%）。结论：我们能够通过分析粪便DNA中的三个遗传标记来检测大多数结直肠癌。需要进一步的工作来确定这些基因检测在无症状患者中检测结直肠肿瘤的特异性，并更准确地估计突变的发生率和检测的敏感性。
Background: Colorectal cancer cells are shed into the stool, providing a potential means for the early detection of the disease using noninvasive approaches. Our goal was to develop reliable, specific molecular genetic tests for the detection of colorectal cancer in stool samples. Methods: Stool DNA was isolated from paired stools and primary tumor samples from 51 colorectal cancer patients. Three genetic targets-TP53, BAT26, and K-RAS-were used to detect tumor-associated mutations in the stool prior to or without regard to the molecular analyses of the paired tumors. TP53 gene mutations were detected with a mismatch-ligation assay that detects nine common p53 gene mutations. Deletions within the BAT26 locus were detected by a modified solid-phase minisequencing method. Mutations in codons 12 and 13 of K-RAS were detected with a digital polymerase chain reaction-based method. Results: TP53 gene mutations were detected in the tumor DNA of 30 patients, all of whom had the identical TP53 mutation in their stools. Tumors from three patients contained a noninherited deletion at the BAT26 locus, and the same alterations were identified in these patients' stool specimens. Nineteen of 50 tumors tested had a KRAS mutation; identical mutations were detected in the paired stool DNA samples from eight patients. In no case was a mutation found in stool that was not also present in the primary tumor. Thus, the three genetic markers together detected 36 (71%) of 51 patients (95 % confidence interval [CI] = 56% to 83%) with colorectal cancer and 36 (92%) of 39 patients (95% CI = 79% to 98%) whose tumors had an alteration. Conclusion: We were able to detect the majority of colorectal cancers by analyzing stool DNA for just three genetic markers. Additional work is needed to determine the specificity of these genetic tests for detecting colorectal neoplasia in asymptomatic patients and to more precisely estimate the prevalence of the mutations and sensitivity of the assay.