通过检测粪便中人类DNA改变进行结直肠癌筛查：多靶点检测试剂盒的可行性

Colorectal cancer screening by detection of altered human DNA in stool: feasibility of a multitarget assay panel.

作者信息

Ahlquist D A, Skoletsky J E, Boynton K A, Harrington J J, Mahoney D W, Pierceall W E, Thibodeau S N, Shuber A P

机构信息

Division of Gastroenterology and Hepatology, Department of Biostatistics, and Division of Molecular Genetics, Mayo Clinic, Rochester, Minnesota 55905, USA.

出版信息

Gastroenterology. 2000 Nov;119(5):1219-27. doi: 10.1053/gast.2000.19580.

DOI:10.1053/gast.2000.19580

PMID:11054379

Abstract

BACKGROUND & AIMS: Assay of altered DNA exfoliated into stool represents an intriguing approach to screen for colorectal neoplasia, but multiple markers must be targeted because of genetic heterogeneity. We explored the feasibility of a stool assay panel of selected DNA alterations in discriminating subjects with colorectal neoplasia from those without.

METHODS

Freezer-archived stools were analyzed in blinded fashion from 22 patients with colorectal cancer, 11 with adenomas > or =1 cm, and 28 with endoscopically normal colons. After isolation of human DNA from stool by sequence-specific hybrid capture, assay targets included point mutations at any of 15 sites on K-ras, p53, and APC genes; Bat-26, a microsatellite instability marker; and highly amplifiable DNA.

RESULTS

Analyzable human DNA was recovered from all stools. Sensitivity was 91% (95% confidence interval, 71%-99%) for cancer and 82% (48%-98%) for adenomas > or =1 cm with a specificity of 93% (76%-99%). Excluding K-ras from the panel, sensitivities for cancer were unchanged but decreased slightly for adenomas to 73% (39%-94%), while specificity increased to 100% (88%-100%).

CONCLUSIONS

Assay of altered DNA holds promise as a stool screening approach for colorectal neoplasia. Larger clinical investigations are indicated.

摘要

背景与目的

检测粪便中脱落的异常DNA是一种用于筛查结直肠肿瘤的有趣方法，但由于基因异质性，必须针对多种标志物。我们探讨了一个选定DNA改变的粪便检测组合在区分结直肠肿瘤患者与非肿瘤患者方面的可行性。

方法

以盲法分析了22例结直肠癌患者、11例腺瘤直径≥1 cm的患者以及28例结肠镜检查正常患者的冷冻保存粪便。通过序列特异性杂交捕获从粪便中分离出人类DNA后，检测靶点包括K-ras、p53和APC基因15个位点中的任何一个的点突变；微卫星不稳定性标志物Bat-26；以及高度可扩增的DNA。

结果

所有粪便均回收了可分析的人类DNA。癌症的敏感性为91%（95%置信区间，71%-99%），腺瘤直径≥1 cm的敏感性为82%（48%-98%），特异性为93%（76%-99%）。从检测组合中排除K-ras后，癌症的敏感性不变，但腺瘤的敏感性略有下降至73%（39%-94%），而特异性提高到100%（88%-100%）。