鲎试剂的平衡浊度值的研究

Study on the equilibrium turbidimetric value of tachypleus amebocyte lysate

目的:研究鲎试剂的平衡浊度值的意义和作用。方法:在 BET-32C 细菌内毒素测定仪上,采用不同的鲎试剂浓度与内毒素进行反应,根据其反应过程中浊度值的变化,获得相应的平衡浊度值和特征反应时间 t_(0.077),研究二者之间关系。其中t_(0.077)表示浊度值达到0.077的时间,也是反应体系形成凝胶所需的时间。结果:(1)4个厂家相同灵敏度的19个鲎试剂,其t_(0.077)的平均 RSD 为25%左右,平衡浊度值为0.087～0.27,主要在0.10～0.20间,差别较大;(2)鲎试剂浓度影响着 t_(0.077)的大小;(3)平衡浊度值与鲎试剂浓度呈线性关系,R^2>0.90;浓度为0.1～1.0 EU·mL^(-1)的细菌内毒素与鲎试剂反应,其平衡浊度值为0.136±0.017;(4)平衡浊度值的变化影响 t_(0.077)值的大小,在试验范围内,t_(0.077)有最小值;对 A、B 2个厂家的6种鲎试剂进行了实验,当平衡浊度值为0.10～0.15时,t_(0.077)最小。结论:鲎试剂的平衡浊度值与凝胶形成的快慢有关,建议通过控制平衡浊度值以充分发挥鲎试剂的效率。

Objective ：To study the meaning of the equilibrium turbidimetric value（ODequi） and its usage in gel- clot assay. Methods ： Endotoxins were reacted with tachypleus amebocyte lysate （TAL） of different concentrations in BET- 32C bacterial endotoxin tester. Then the ODequi ,characteristic reactive time t0.077 were read and the relationship between them was investigated. The t0. 077, which was defined as the time spent when the turbidimetric value of the mixture reached to 0. 077, indicates the time needed when the gel - clot formed in the mixture. Results ： （ 1 ） The mean RSD of t0.077 of the same sensitivity TALs from four manufacturers was about 25% and their ODequi ranged from 0. 087 to 0. 27, mainly from 0. 10 to 0. 20 ; （2） The t0. 077 were changed with the TAL concentrations ; （ 3 ） The ODequi were proportioned to the TAL concentrations. The ODequi of the tested TAL was 0. 136 ±0. 017 when 0. 1 - 1.0 EU · mL^-1 endotoxin were used; （4） The minimum t0.077 were observed when the ODequi were from 0. 10 to 0. 15 for TALs of manufacturer A and B. Conclusion：The equilibrium turbidimetric values, read by the dynamic turbidimetric method in bacterial endotoxin tester, were proportioned to the TAL concentrations and less affected by the endotoxin concentration. The minimum t0.077 was observed when the ODequi changed.