重组链激酶活性测定中图片上溶酶圈直径测量方法

目的: 研究建立在图片上测量重组链激酶溶解圈直径的方法。 方法: 在图片上溶解圈边缘人工取点(大于3个),利用最小二乘法,回归计算圆方程,并编写了相应的计算软件(circular_regression),并对所建方法的各项指标进行验证。通过在不同人员,在不同时间里的重复测量来评价软件所得数据的准确性、线性、重复性和精密度。通过在不同电脑上安装该软件来评估同图备份中同一溶解圈直径,进行软件粗犷度分析。用所建方法与直接测量方法进行比较。 结果: 在对参考圆的测定中,实测值与理论值之间差小于理论值的1%,并且所得数值的线性好,实测值与理论值之间的相关系数>0.9999。从重复性和精密度上看,试验结果测量的RSD<1%。从软件的粗犷度上看,2台电脑的分析结果无差别。所建方法测量结果与直接测量方法测量结果之间无显著性差异。 结论: 该自制软件具有准确性高,精密度高,重复性好,线性好等特点,可以用于替代直接测量方法,用于重组链激酶活性检测分析。

Objective:To establish methods to determine the diameter of transparent lysis ring in pictures derived from biological activity tests for recombinant streptokinase and to valid.Method:The diameters of transparent lysis rings in the picture derived from biological activity test of recombinant streptokinase were determined by software "circular_regression" programmed by ourselves.Accuracy and linearity was assessed by analyzing the diameters of a series of reference circles in the specific standard picture for the software.Intermediate precision and repeatability was assessed by repeating analysis of three transparent lysis rings in same picture by three different analysts in three different days.Ruggedness was assessed by comparing the measuring results from different computers installed with the same software.Results:The results of the accuracy and linearity study indicated that the differences between reference diameters and measuring diameters were all less than 1%.The r between reference diameters and measuring diameters was higher than 0.9999.The results based on precision and repeatability study showed the RSD of the measuring diameters was less than 1%.There was no significant difference of the measuring diameters of the transparent lysis ring in the picture copies between two computers.Conclusion:Results from the validation study demonstrated that software measurement is accurate,precise,and with good repeatability and linearity for measuring the diameters of transparent lysis rings.The method could be an alternative of measurement for transparent lysis rings.

1 ChP(中国药典).2005 Vol Ⅲ(三部).Appendix(附录) Ⅵ B
2 Bates DG,Cosentino C.Validation and invalidation of systems biology models using robustness analysis.IET Syst Biol,2011,5(4):229
3 Beck F,Lewandrowski U,Wiltfang M,et al.The good,the bad,the ugly:validating the mass spectrometric analysis of modified peptides.Proteomics,2011,11(6):1099
4 Strino F,Parisi F,Kluger Y.VDA,a method of choosing a better algorithm with fewer validations.PLoS One,2011,6(10):e26074
5 ZHANG Yi(张翊),DING You-xue(丁有学),RAO Chun-ming(饶春明),et al.The study of national reference material of rSK(重组链激酶(rSK)国家标准品的研制).Chin J Biol(中国生物制品学杂志),2000,13(4):46