实验误差因素对人双核淋巴细胞微核结果影响的研究

Effects of Experiment Error Factors on Micronucleus Frequency Scoring in Cytokinesis-Blocked Lymphocyte

背景与目的:探讨不同染色时间、不同阅片时间及阅片人的熟练程度对微核识别率的影响。材料与方法:血样采自1名30岁健康男性。分离淋巴细胞并以RPMI1640培养,分2个照射组(1Gy、2Gy)和1个对照组,共3组。采用胞质分裂阻滞微核实验方法低渗制片,每组制备2张玻片,每张玻片滴制a、b2个计数点,并分2批在不同时间用Giemsa染色。6名对微核实验熟练程度不同的阅片人分别按要求阅片。结果:在统一计数标准的前提下,阅片人熟练程度导致的微核判读结果差异不显著(P>0.05);全部阅片人在不同时间对同一玻片a、b2个计数点的微核计数结果之间差异无统计学意义(P>0.05);不同时间染色标本的核分裂指数(NDI)、微核细胞数(MNed)、微核数(MNi)和核分裂指数(NDI)等结果之间差异无统计学意义(P>0.05)。结论:在本实验条件下,阅片人熟练程度、不同时间染色或阅片均不会对微核识别的判断造成明显影响。

BACK（ROUND ＆ AIM：To study the effect on MN frequency identification caused by different skill level, staining time and scoring time. MATERIALS AND METHODS： A blood sample was collected from a healthy 30-year-old male. Lymphocytes was isolated and then cultured in RPMI medium in triplicates. Two of the 3 cultured groups were then exposed to 1Gy or 2 Gy gamma-rays and the other one was control. Parallel scoring spots （a and b） in same slide was made according to CBMN assay method, 2 simultaneously parepared slides of each group was stained by Giemsa at 2 different times, and then 6 scorers with different skill levels scored the MN frequency. RESULTS： Variation of MN frequency scored by scorers with different skill levels was not significant. The MN frequency scoring for spot a and b at different times showed no significant difference among different scorers. The observed endpoints, MNed, MNi, NPB and NDI, stained at different times, were not different significantly either. CONCLUSION： Skill level, staining time or scoring time did not show significant interference on estimation of the dose-effect relationship between MN frequency and radiation dose.