辐射诱导染色体畸变的快速FISH方法的建立

目的： 建立一种检测辐射诱导染色体畸变的快速荧光原位杂交(FISH)方法。方法：两步简并引物PCR法扩增人α-卫星DNA，制备荧光基团直接标记的泛着丝粒探针；对60Coγ射线照射后的人外周血淋巴细胞染色体标本进行FISH分析，荧光显微镜下检测着丝粒及染色体形态。结果：直接标记泛着丝粒探针杂交后显示所有染色体着丝粒均有较强的信号；应用制备的FISH探针检测到γ射线照射诱导的双着丝粒、着丝粒环和易位染色体畸变，37 ℃杂交5-12 h后经过简单的洗涤即可在荧光显微镜下检测到较好的信号。结论：本研究制备的直接标记泛着丝粒探针，可用于FISH快速检测辐射诱导的染色体双着丝粒体、着丝粒环和易位畸变。

method to analyze chromosomeaberrations induced byirradiation

OBJECTIVE： To establish a rapid fluorescence in situ hybridization （FISH） method and to analyzethe chromosome aberrations induced by irradiation. METHODS：Human centromeric alpha satellite DNA was amplifiedand directly labeled with Cy3-dUTP or Fluorescein-12-dUTP by degenerate oligonucleotide priming-PCR （DOP-PCR） toprepare two pan-centromeric probes. Chromosome aberrations in lymphocytes of healthy irradiated peripheral blood60 samples with Coγ-rays were analyzed with FISH. RESULTS：Two directly labeled pan-centromeric probes hybridizedto the centromeres of all 46 chromosomes. The established rapid FISH method could easily detect chromosome aberrationsafter hybridizing 5 h to 12 h in 37 ℃ and simple washing. CONCLUSION： FISH method established in this study couldbe use to rapidly detect the dicentrics,centric rings,and translocations induced by irradiation.