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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[革兰氏阴性菌:细菌内毒素(0.0625,0.125,0.25,0.5,1.0 EU.mL-1);真菌:酵母多糖(2.0 mg.mL-1);革兰氏阳性菌:脂磷壁酸(0.5μg.mL-1)]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0.5μg.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 study the feasibility of detecting the pyrogen by rabbit whole blood in vitro. Methods: Bacterial endotoxin test water (negative control), three kinds of pyrogen solution [Gram-negative bacteria: bacterial endotoxin (0.0625,0.125,0.25,0.5,1.0 EU.mL-1), fungi : Zymosan (2.0 mg.mL-1); Gram-positive bacteria: Lipoteichoic acid (0.5 μg.mL-1)] were added into whole rabbit blood and incubated in a CO2 incubator (37 ° C, 5% 8 h. The concentration of cytokines (IL-6) in the incubation system was detected by enzyme-linked immunosorbent assay (ELISA), and the content of pyrogen was indicated by IL-6 concentration. Results: The lowest limit of detection (LOD) of bacterial endotoxin, zymosan and lipoteichoic acid was 0.5 EU.mL-1, 2.0 mg.mL-1 and 0.5 μg.mL-1 respectively. France’s minimum detection limit are greater than the rabbit law. Conclusion: In vitro rabbit whole blood method is not suitable for the detection of pyrogen by using cytokines (IL-6) as the detection index.