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目的对Pig-a基因突变试验设计和统计学分析方法提出建议。方法使用本实验室阴性对照历史数据128例,使用描述统计学获得数据分布特征。在此基础上对不同条件(检测细胞数目、每组动物数量、试验组数)进行模拟试验,提出可达到理想统计学功效的试验设计建议和统计学分析方法。结果阴性样本突变成熟红细胞(RBCsCD59-)和突变网织红细胞(RETsCD59-)均数为26.63×10-6、35.85×10-6,标准差为27.71×10-6、31.06×10-6。频率分布图显示RBCsCD59-和RETsCD59-呈偏态分布,经对数转换后呈正态分布。在精度为2个标准差时,每样本1×106个RBCs和0.3×106个RETs的总体均数置信度分别为100%和92%;若RETs的检测数量扩大至1×106时,置信度可达100%。当检测最小差异为2倍时,使用每组5只动物的统计学功效均较低;当检测最小差异为4倍或5倍时功效为>80%。结论本实验室对Pig-a基因突变时试验设计建议如下:①建议将数据进行log(10)转化后可进行参数检验,如方差分析。②建议选用年轻成年动物进行试验,大鼠约为6周龄。③进行流式细胞术检测时建议获取细胞量为:RETs>1×106,RBCs>5×107。④检测差异为4倍以上时,使用3个剂量组和1个阴性对照组可获得理想的统计学功效。⑤在剂量组为4组时(包括对照组),每组5只动物可基本满足试验要求,但为获得更良好的结果推荐使用的动物数量为每组6只或7只。
Abstract：Objective To make recommendations on the design and analysis of Piga gene mutation assay on the basis of the historical control data.Methods All negative historical control data( total: 128) were collected to create the historical control data base,and descriptive statistics was used to determine the distribution of the data.The power of the number of animals per group,number of measured cells per animal and number of groups were discussed through simulation test based on historical data base.Results The group mean of mutant red blood cells( RBCsCD59-) and mutant reticulocytes ( RETsCD59-) were 26.63 × 10-6 and 35.85 × 10-6,respectively,and the standard deviation were 27.71 × 10-6 and 31.06 × 10-6,respectively.By mapping the frequencies of data,the variables had skewed distributions and translated to normal distributions after logarithmic transformation.The confidence level for population means were 100% and 92% for RBCs( 1 × 106 cells per animal) and RETs( 0.3 × 106 cells per animal),and it increased to 100% when 1 × 106 cells was scored for RETs.Group sizes of 5 had low statistical power while the minimal detectable difference was 2 times.The power had increased to > 80% when 4- or 5- fold of minimal detectable difference was employed.Conclusion In brief,the recommendations include:① A log( 10) transformation of mutant frequencies often has been found satisfactory for data when parametric method such as an analysis of variance are used.② Young adult animals( approximately 6 weeks of age rats) are recommended in this assay.③ The recommended number of RETs and RBCs are > 1 × 106 and > 5 × 107,respectively.④ Based on power calculations,3 treatment groups and 1 negative control group are appropriate when fourfold increase was employed.⑤ Group sizes of 6 or 7 are recommended but 5 analyzable animals per group may be acceptable when 4 test groups were used.
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基本信息:
DOI：10.19813/j.cnki.weishengyanjiu.2018.04.002
中图分类号:R346
引用信息:
[1]霍娇,刘运杰,游一屏,等.大鼠体内Pig-a基因突变试验设计及统计学分析方法建议[J].卫生研究,2018,47(04):525-529+553.DOI:10.19813/j.cnki.weishengyanjiu.2018.04.002.
基金信息:
国家自然科学基金面上项目(No.8157120809)
2017-10-25
2017
2017-12-28
2018-01-24
2018
1
2018-07-27
2018-07-27