应用荧光多重聚合酶链反应法检测结直肠癌微卫星不稳定

目的建立一种敏感、稳定、高通量的结直肠癌微卫星不稳定(MSI)检测技术。方法105例散发性结直肠癌患者取其新鲜癌组织及正常肠黏膜,DNA提取,荧光多重聚合酶链反应(PCR)扩增5个微卫星位点,应用GeneScan方法分析PCR产物。结果105例结直肠癌患者26例(24.7%)存在微卫星不稳定,其中MSI-H 14例(13.3%),MSI-L 12例(11.4%),MSS 79例(75.3%)。D5S346、BAT26、BAT25、D17S250、D2S123突变率分别为5.6%、8.6%、10.5%、8.6%、10.5%。结论应用荧光多重PCR可以高通量检测结直肠癌微卫星不稳定,敏感性高,结果可靠,适合临床应用。

The detection of microsatellite instability in colorectal cancer by fluorescence multiplex PCR

Objective To set up a sensitive and stable technique which has high throughout to detect the microsatellite instability of colorectal cancer. Methods Genomic DNA extracted from the cancer tissues and their normal tissues were subjected to microsatellite instability analysis on 5 microsatellite loci in 105 cases of sporadic colorectal cancer by means of fluorescence multiplex polymerase chain reaction and fragment analysis methods of GeneScan. Results All 5 microsatellite markers were amplified in 105 cases. Of the 105 cases,there were 14 (13.3%) cases of MSI-H,12 (11.4%) cases of MSI-L and 79 (75.3%) cases of MSS. Instability of D5S346 was found in 6 cases (5.6%) , BAT-26 in 9 cases (8.6%) ,BAT-25 in 11 cases (10.5%) , D17S250 in 9 cases (8.6%) and D2S123 in 11 cases ( 10. 5% ) respectively. Conclusion Using fluorescence multiplex polymerase chain reaction to detect the microsatellite instability of colorectal cancer is an effective, stable and sensitive method and can be used in practice.