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目的 建立多色荧光原位杂交方法 ,并探讨用该方法进行早先辐射受照射者的剂量重建的可行性。方法 筛选 8对染色体端粒和着丝粒特异性人工细菌染色体 (BAC)克隆 ,建立多色荧光原位杂交方法 ,用该方法分析60 Coγ射线离体照射的新生儿脐带血淋巴细胞染色体畸变 ,并建立相应的标准剂量 效应曲线 ,参照标准剂量 效应曲线来估算两例早先受照射者的累积吸收剂量。结果 本研究中建立的多色荧光原位杂交方法用生物素和 (或 )地高辛将端粒和着丝粒BACDNA标记成绿、红、黄 (绿 +红 ) 3种荧光染色 ,使得 1 ,2 ,3 ,7,8,9,1 4和 1 5号染色体很容易辨认。用该方法分析受60 Coγ射线照射的新生儿脐带血淋巴细胞染色体畸变 ,除断片外 ,其他染色体畸变的剂量 效应曲线均为线性二次剂量反应模式。用所有细胞或稳定性细胞中的完全相互易位率作指标 ,估算了两例早先受射照者的吸收剂量。结论 本研究所建立的多色荧光原位杂交方法可以用来进行早先受照射者的剂量重建
Objective To establish a multi-color fluorescent in situ hybridization (FISH) method and to explore the feasibility of using this method to reconstitute the dose of previous irradiation. Methods Eight pairs of chromosomal telomere and centromere-specific artificial bacterial chromosome (BAC) clones were screened to establish a multicolor fluorescence in situ hybridization method. Chromosome aberrations of neonatal umbilical cord blood lymphocytes irradiated by 60 Co γ -ray were analyzed by this method. And establish the corresponding standard dose-response curve, with reference to the standard dose-response curve to estimate the cumulative absorbed dose of two previously irradiated subjects. Results The multi-color fluorescence in situ hybridization method established in this study labeled telomere and centromeric BAC DNA with green fluorescent dye, red fluorescent dye and green fluorescent dye with digoxin, making 1 2, 3, 7, 8, 9, 14 and 15 chromosomes are readily identifiable. This method was used to analyze the chromosomal aberrations of neonatal umbilical cord blood lymphocytes irradiated by 60 Co γ-rays. The dose-response curves of other chromosomal aberrations were linear quadratic dose response patterns except fragment. Using the complete reciprocal translocation rate in all cells or stable cells as an index, the absorbed dose of two previously irradiated subjects was estimated. Conclusions The multi-color fluorescence in situ hybridization method established in this study can be used to perform dose reconstruction of previously irradiated subjects