建立可吸收医用膜细菌内毒素检查的新方法

[目的]检验粘克^TM可吸收医用膜是否符合生物材料的热原要求,建立粘克^TM可吸收医用膜的细菌内毒素检查方法。[方法]按《中华人民共和国药典》2005年版附录及GB/16175-1996规定,同时采用热原检查法和细菌内毒素检查法对粘克^TM可吸收医用膜72h的浸提液样品进行检测。[结果]该浸提液对细菌内毒检测不产生干扰,细菌内毒素试验结果为阴性。同时受试家兔体温升高均低于0.6℃,并且3只家兔体温升高总和低于1.4℃。[结论]粘克^TM可吸收医用膜不含引起机体发热的内毒素或致热原,符合医用生物材料的使用标准。用细菌内毒素检查法替代家兔热原法对该可吸收膜进行热原检查是可行的。     

透析用水细菌及内毒素含量在热消毒和化学消毒全过程中的动态规律特征研究

 目的 观察透析用水中细菌存活及内毒素含量在两种消毒过程中的动态变化情况。利用数据模型拟合消毒过程中微生物指标变化规律,为日常感染监控及管路消毒维护管理决策提供基础数据支撑。方法 分析消毒程序模式的时间和温度参数,采集热消毒和化学消毒前后输水管路回水口或混合室入口处的透析用水,检测内毒素,同时平行水样经薄膜过滤法富集,在20℃下TGEA培养7 d,计数细菌菌落。通过Origin软件的数据拟合,描述细菌和内毒素含量与温度、时间变化的特征关系。结果 细菌存活率随着消毒程序时间的延长而减少,在50~160 min时,细菌存活情况变化幅度大,热消毒细菌含量减少的变化比化学消毒平缓。热消毒中内毒素含量随着温度不断上升而增加,随着冷却温度下降而减少。在70、80℃时内毒素含量最高,首机处内毒素含量整体水平高于中间机和尾机。化学消毒内毒素含量集中升高在消毒剂浸泡冲洗时间段,之后下降而趋于平稳。热消毒和化学消毒完内毒素持续情况均为先下降到较低值,在热消毒后15 h、化学消毒后12 h,内毒素略呈现上升趋势,出现小峰值。结论 温度和时间是影响透析用水微生物指标决定性的参数,可利用微生物数据建立拟合数据描述两种消毒全过程透析用水中细菌失活和内毒素变化规律的异同。同时模型工具补充了透析用水监测方法,有助于更好地实现透析用水消毒控制及效果。     

鲎试剂法检测采血原辅材料细菌内毒素的应用

细菌内毒素是微生物的代谢产物,是引起热原反应的主要物质。鲎试剂为鲎血变形细胞裂解物的冻干精制品,含有能被内毒素激活的凝固系统,在一定的条件下极微量的细菌内毒素即可激活鲎试剂中的C因子,最终形成肉眼可见的凝胶。鲎试剂法检测细菌内毒素以其快速、简便、灵敏度高、重现     

输液连接件 ( 肝素帽 ) 可沥滤物检测浸提条件的确定

通过各种标准中浸提比例的比较和实际产品表面积的测量，确定浸提比例为1克样品加20m l蒸馏水。在不同浸提条件下，对产品化学性能（还原物质、酸碱度、重金属、紫外吸光度）测定结果进行了比较,确定最佳浸提条件为37＆#177;1？C,24h。     

葛根素注射液细菌内毒素检查方法的研究

目的:考察葛根素注射液细菌内毒素检查方法的可行性.方法:依据<中国药品检验标准操作规程>2000年版细菌内毒素检查法,确定葛根素注射液的有效稀释浓度和细菌内毒素限值.结果:葛根素注射液在最大有效稀释浓度下对鲎试剂无干扰作用,其内毒素限值符合规定.结论:可用细菌内毒素检查法取代<中国药典>2000年版(二部)规定的热原检查法.     

鲎试剂用于20%甘露醇注射液细菌内毒素检测

目的:研究甘露醇注射液细菌内毒素的检查方法。方法:按照《中国药典》2000年版(二部)附录方法和指导原则。结果:甘露醇注射液在最大有效稀释浓度下对鲎试剂反应无干扰作用。结论:可用细菌内毒素检查法取代热原检查法。     

生物学试验中不同浸提条件下镍钛合金中镍溶出的研究

研究镍钛合金医疗器械在进行生物学试验样品制备时,不同浸提条件下制备的浸提液中镍溶出情况,分析不同浸提条件之间是否具有等效性,找到最佳的浸提条件。该实验选用两种品牌的镍钛合金材料,按GB/T16886-12中推荐的浸提条件对材料进行浸提,采用等离子发射光谱仪对浸提液中镍浓度进行分析。结果发现对于镍钛合金材料,各浸提条件间不具有等效性,建议常规浸提选用(37±1)℃,(72±2)h,加速浸提可选用(70±2)℃,(24±2)h。     

安必洁多酶清洗液清除细菌内毒素实验研究

目的探讨安必洁多酶清洗液清除细菌内毒素效果的可行性. 方法将被人工污染细菌内毒素的玻璃注射器浸泡在不同浓度、时间的多酶清洗液中至规定时间,然后按GB/T14233.2-1993行细菌内毒素检测,并与三效热原灭活剂清除效果比较. 结果安必洁多酶清洗液1∶200作用30min、1∶150作用10min、1∶100作用5 min细菌内毒素检测全部阴性;与800 mg/L三效热原灭活剂作用60 min效果相当. 结论安必洁多酶清洗液具有快速清除细菌内毒素的效果.     

人全血IL-1β试验在医疗器械体外热原检测中的应用前景探讨

目的:探讨人全血IL-1β试验在医疗器械热原体外检测中应用的可行性。方法:该研究采用梯度稀释的内毒素O113与人新鲜全血孵育,检测上清所含IL-1β的量并建立标准曲线;采用脂磷壁酸(LTA)溶液与新鲜人全血孵育,检测上清所含IL-1β的量并根据回归方程计算出LTA溶液的内毒素当量。结果:成功建立标准曲线,并计算出LTA溶液的内毒素当量。结论:经分析比较,该研究认为人全血IL-1β试验可用于医疗器械热原检测并能弥补传统方法的诸多不足。     

血必净注射液细菌内毒素检查法的研究

目的:建立血必净注射液的细菌内毒素检查法.方法:将血必净注射液经12倍稀释,用标示灵敏度为0.25EU·ml-1的鲎试剂检测其细菌内毒素.结果:血必净注射液稀释12倍对鲎试剂无干扰作用.结论:可以用细菌内毒素检查法对血必净注射液进行热原检查.     

蓝域染色剂的细菌内毒素检测

目的:考察蓝域染色剂的细菌内毒素检查方法的可行性.方法:依据<中华人民共和国药典>2005年版附录XIE细菌内毒素检查法,确定蓝域染色剂的有效稀释浓度和细菌内毒素限值.结果:蓝域染色剂在稀释16倍后对细菌内毒素实验无干扰作用,其细菌内毒素限值符合规定.结论:该产品可以采用细菌内毒素检查法.     

参麦注射液细菌内毒素检查方法的探讨

目的：考察参麦注射液细菌内毒素检查方法的可行性,加强对参麦注射液的质量监控。方法：采用细菌内毒素检查法,考察参麦注射液对鲎试剂的干扰试验,并检测供试品细菌内毒素的限度。结果：对样品进行1∶4稀释,用0.125 EU/mL鲎试剂无干扰,与家兔热源检查法结果一致。结论：与家兔法检查本品热原相比,细菌内毒素法可行性高,此方法避免了家兔热源检查法操作的复杂性,且灵敏度高、方便快捷。     

枯痔注射液细菌内毒素检查方法研究

目的研究枯痔注射液的细菌内毒素检查法。方法依照《中国药典》2010年版(二部)附录方法和指导原则。结果枯痔注射液在最大有效稀释浓度下对鲎试剂反应无干扰作用。结论用细菌内毒素检查法对枯痔注射液进行热源检查是可行的。     

动态浊度法测定人凝血酶原复合物制品中细菌内毒素含量

[目的]建立动态浊度法定量测定人凝血酶原复合物制品中细菌内毒素含量的方法。[方法]通过对样品中定量添加标准内毒素的干扰试验,检测其回收率应在50%～200%范围。[结果]人凝血酶原复合物制品对鲎试验无干扰作用,动态浊度法定量检测结果与家兔实验结果基本一致。[结论]应用本方法定量测定人凝血酶原复合物制品中细菌内毒素含量是可行的。     

The role of fever in appetite suppression after endotoxin administration

In order to test the hypothesis that fever, and not some other aspect of the acute phase response, decreases food intake after administration of endotoxin, food intake of rats was studied under conditions of 1) fever, 2) antipyresis, and 3) endotoxin tolerance. Injection of endotoxin resulted in a significant elevation in rectal temperature and a significant reduction in food intake. Administration of the antipyretic drug sodium salicylate to endotoxin-injected animals lowered rectal temperatures to control levels, but food intake was still suppressed. When rats were made tolerant to endotoxin by repeated injections, an attenuated fever was observed, and food intake was not significantly different from that of control animals. We conclude that the effects of endotoxin on body temperature can be dissociated from its effects on food intake. We speculate that the failure of endotoxin to suppress food intake in endotoxin-tolerant rats may be due to a decreased production of endogenous pyrogen.     

In vitro pyrogenicity of the diphtheria, tetanus and acellular pertussis components of a trivalent vaccine

We have earlier found that a trivalent vaccine, containing antigenic components from both Gram-positive and Gram-negative bacteria, induced secretion of the endogenous pyrogen interleukin 6 (IL-6) when added to fresh human blood in vitro. The results of the present study showed that the IL-6 secretion was induced by toxoids derived from the Gram-positive bacterium Corynebacterium diphtheriae. However, fresh whole blood from different donors reacted differently to the stimulation. The blood from some donors induced secretion of large concentrations of IL-6, while the blood from other donors induced essentially no IL-6 secretion as a response to stimulation with diphtheria toxoid or a mixture of diphtheria and tetanus toxoids. Repeated testing over several years using blood from the same donor confirmed a donor-dependency of the reaction. This donor-dependency was only found for the toxoid, since blood from all donors reacted with approximately similar IL-6 production to stimulation by endotoxin from the Gram-negative bacterium Escherichia coli, known to be mediated via the toll-like receptor (TLR) 4. Also, no donor-dependecy was found to highly purified lipoteichoic acid from the Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus, known to be mediated via TLR-2 and TLR-6. The receptors involved in stimulation by diphtheria toxoid are not known, but may differ from those used by endotoxin and lipoteichoic acid.     

钛镍记忆合金组织吻合器细菌内毒素检测方法的研究

目的：对钛镍记忆合金组织吻合器进行细菌内毒素检查,建立其细菌内毒素检查方法。方法：根据《中国药典》2005版二部附录收载的细菌内毒素检查方法,使用鲎试剂对钛镍记忆合金组织吻合器进行干扰试验。结果：钛镍记忆合金组织吻合器可用标示灵敏度为0.25EU/mL的鲎试剂进行细菌内毒素检查。结论：该方法准确、可靠,具有良好的重复性和灵敏度,可用于钛镍记忆合金组织吻合器细菌内毒素的测定。     

Monocyte-activation test to reliably measure the pyrogenic content of a vaccine: An in vitro pyrogen test to overcome in vivo limitations

Pyrogen content is one of the critical quality attributes impacting the safety of a product, and there is an increasing need for assays that can reliably measure this attribute in vaccines. The Limulus amebocyte lysate (LAL) assay and the rabbit pyrogen test (RPT) are the canonical animal-based pyrogen tests currently used to release vaccines; however, there are several drawbacks associated with these tests when applied to Bexsero, intrinsically pyrogenic product, containing a meningococcal Outer Membrane Vesicle component. While the RPT, as applied to Bexsero at its given dilution, ensures safe vaccine, it is highly variable and prone to false positive results. On the other hand, the LAL assay although quantitative, can detect only endotoxin pyrogens and is not sufficient for monitoring the safety of Bexsero, which contains both LPS and non-endotoxin pyrogens. Being aware of these limitations of the RPT and LAL when applied to Bexsero, the Monocyte Activation Test (MAT) which is sensitive to both endotoxin and non-endotoxin based pyrogens has been developed as an alternative pyrogen test. Here, the development and the validation of a MAT assay adapted from the European pharmacopoeia for Bexsero, is described. The MAT assay is then used for monitoring the safety and consistency of Bexsero vaccines at release, providing great advantages in terms of reduced variability with respect to RPT, reduction of animal use, in line with the 3Rs principle concerning the protection of animals and faster time to market. In addition the correlation of the MAT to the RPT has been demonstrated supporting the replacement of the in vivo method and the potential application of the assay to other intrinsically pyrogenic vaccines.     

依替巴肽注射液细菌内毒素方法研究

目的：对依替巴肽注射液的细菌内毒素检查方法进行探讨,建立其细菌内毒素检查方法。方法：按《中国药典2010》二部附录Ⅺ E细菌内毒素检查方法进行。结果：在验证条件下,依替巴肽注射液可以采用稀释方法排除干扰。结论：可以采用此法对本品的细菌内毒素进行检查。