新的体外热原试验方法研究进展

The progress of a new in vitro pyrogen test

非肠道给药的药品、生物制品及植入性医疗器械要求无热原,目前各国药典采用兔热原试验或细菌内毒素检查法(BET)这2种基于动物的方法进行试验,但它们还存在一些问题。外热原可刺激人单核细胞释放炎性因子(如 IL-1B,IL-6,TNF-α,IFN-λ等)即内热原,内热原使人体温调定点升高从而引起体温升高,这些内热原的含量可用 ELISA 方法进行测定,新的体外热原试验方法是基于此原理的系统。新方法结合了 BET 高灵敏度和家兔法宽检测谱的优点,与兔法相比,新方法更加灵敏、费用低,能直接反映人对多种热原的反应,与细菌内毒素检查法相比,新方法不仅仅局限于革兰氏阴性菌产生的细菌内毒素的检测,对各种热原质均能检测。在欧洲,目前有6种最突出的系统被确认,试验表明他们是安全的、不使用动物的、更有效的热原检查方法,可以作为热原检查替代方法向药政管理部门和药典推荐。

It is a requirement that parenteral medicines or biological products and embedded medical devices be free of pyrogens. All pharmacopoeia use one of two animal - based tests ： the rabbit pyrogen test and the BET present, but there are some problems to them. When human monocytic cells are stimulated by exogenous pyrogens in circulation, they will release proinflammatory cytokines （such as IL-1β,IL-6,TNF-α,IFN-λ, et al）i. e. endogenous pyrogens. These cytokines transmit signals to increase the thermostatic set point and trigger to increase the body temperature, the content of cytokines can be measured using ELISA. The new in vitro pyrogen test system is based on that. The new method combines the high sensitivity of the BET with the wide range of pyrogens detectable by the rabbit pyrogen test. In comparison with the rabbite pyrogen test, the new method is more sensitive, less expensive and reflects the human reaction to the pyrogen directly. Compared to bacterial endotoxin test, the new method is not restricted to endotoxins of gram - negative bacteria. In Europe, there are six most prominent test systems were formally validated,evidence indicated they are safer, animal -free and more efficient pyrogen detection system and they are suitable for introduction to regulatory acceptance and pharmacopoeia.