针对结直肠组织样本中少量受损DNA进行优化的微卫星不稳定性诊断引物组。
Diagnostic primer sets for microsatellite instability optimized for a minimal amount of damaged DNA from colorectal tissue samples.
作者信息
Umetani N, Sasaki S, Watanabe T, Ishigami H, Ueda E, Nagawa H
机构信息
Department of Surgical Oncology, School of Medicine, University of Tokyo, Japan.
出版信息
Ann Surg Oncol. 2000 May;7(4):276-80. doi: 10.1007/s10434-000-0276-6.
BACKGROUND
The diagnosis of microsatellite instability from a minimal amount of highly damaged DNA, extracted from formalin-fixed, paraffin-embedded tissue by the microdissection method, is difficult. Therefore, optimized primer sets were newly designed for substitution of documented ones.
METHODS
DNA was extracted from 15 archival colorectal carcinomas and used as templates for polymerase chain reaction. Nine standard microsatellite markers (BAT-25, BAT-26, BAT-40, D18S69, D2S123, D5S346, D10S197, D17S250, and D18S58) were selected for diagnosis of microsatellite instability in colorectal carcinomas. All polymerase chain reaction conditions for primer sets were unified to save experimental time.
RESULTS
The primer sets for the latter five markers documented in the literature were redesigned because of poor efficiency for damaged DNA. As a result, the number of DNA samples, sufficiently amplified at all markers, improved from 0% to 93%.
CONCLUSIONS
Diagnostic primer sets for microsatellite instability, optimized for a minimal amount of damaged DNA from colorectal tissue samples, were established.
背景
从通过显微切割法从福尔马林固定、石蜡包埋组织中提取的少量高度受损的DNA中诊断微卫星不稳定性是困难的。因此，新设计了优化的引物组以替代已记录的引物组。
方法
从15例存档的结肠直肠癌中提取DNA，并用作聚合酶链反应的模板。选择9个标准微卫星标记（BAT-25、BAT-26、BAT-40、D18S69、D2S123、D5S346、D10S197、D17S250和D18S58）用于诊断结肠直肠癌中的微卫星不稳定性。为节省实验时间，统一了所有引物组的聚合酶链反应条件。
结果
由于对受损DNA的扩增效率不佳，重新设计了文献中记录的后五个标记的引物组。结果，在所有标记上均充分扩增的DNA样本数量从0%提高到了93%。
结论
建立了针对结肠直肠组织样本中少量受损DNA进行优化的微卫星不稳定性诊断引物组。
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