Collins A, Dusinská M, Franklin M, Somorovská M, Petrovská H, Duthie S, Fillion L, Panayiotidis M, Raslová K, Vaughan N
Rowett Research Institute, Bucksburn, Aberdeen, UK.
Environ Mol Mutagen. 1997;30(2):139-46. doi: 10.1002/(sici)1098-2280(1997)30:2<139::aid-em6>3.0.co;2-i.
The comet assay (single-cell gel electrophoresis), which measures DNA strand breaks at the level of single cells, is very easily applied to human lymphocytes, and therefore lends itself to human biomonitoring studies. For the examination of DNA base oxidation (a specific marker of oxidative damage), the assay is modified by including a stage at which the DNA is incubated with a suitable lesion-specific endonuclease. Here we report on the reliability and reproducibility of this approach, from the level of comparing results from duplicate gels prepared from the same sample of cells, up to an assessment of the natural intra- and interindividual variability in lymphocyte DNA damage measured in groups of normal, healthy human volunteers. We applied the assay in investigations of human disease and occupational exposure of factory workers.
彗星试验（单细胞凝胶电泳）可在单细胞水平上检测DNA链断裂情况，该方法很容易应用于人类淋巴细胞，因此适用于人体生物监测研究。为检测DNA碱基氧化（氧化损伤的一种特定标志物），可通过增加一个将DNA与合适的损伤特异性核酸内切酶一起孵育的步骤来对该试验进行改良。在此，我们报告了这种方法的可靠性和可重复性，范围从比较同一细胞样本制备的重复凝胶的结果，到评估正常健康人类志愿者群体中淋巴细胞DNA损伤的自然个体内和个体间变异性。我们将该试验应用于人类疾病研究以及工厂工人的职业暴露调查。
