兔全血法检测热原的可行性探讨

目的: 研究体外兔全血法检测热原的可行性。 方法: 分别将细菌内毒素检查用水(阴性对照)、3种来源的热原质溶液 加入兔全血,在CO₂培养箱(37 ℃,5% CO₂)内孵育8 h,用酶联免疫分析法(ELISA)检测孵育体系中细胞因子(IL-6)的浓度,以IL-6浓度指示热原质的含量。 结果: 家兔热原检查法对细菌内毒素、酵母多糖及脂磷壁酸的最低检测限为0.5 EU·mL^-1、2.0 mg·mL^-1及0.5 μg·mL^-1,体外兔全血法的最低检测限均大于家兔法。 结论: 以细胞因子(IL-6)为检测指标,体外兔全血法不适用于热原质的检测。

Objective: To discuss the feasibility of the in vitro pyrogen test based on the rabbit whole blood. Method: Taking rabbit whole blood incubated with water (negative control) and three different sources of pyrogen such as endotoxins (0.0625,0.125,0.25,0.5,1.0 EU·mL^-1) from Gram-negative bacterial,LTA (0.5 μg·mL^-1) from Gram-positive bacterial,zymosan (2.0 mg·mL^-1) from fungi for 8 h (37 ℃,5%CO₂),respectively.Then the concentration of interleukin-6 (IL-6) in the supernatants of the culture medium was measured by ELISA.The concentration of IL-6 was used to indicate concentration of pyrogen and deduced minimum detection limit.Stimultaneously,minimum detection limit was contrasted to the rabbit method's. Results: Minimum detection limit of rabbit method to endotoxins,LTA and zymosan is 0.5 EU·mL^-1,0.5 μg·mL^-1 and 2.0 mg·mL^-1 respectively.But minimum detection limit of rabbit whole blood method is higher than rabbit method's. Conclusion: It is not adaptive for the rabbit whole blood to detect pyrogen which using the IL-6 as the indicator.