救治破伤风抗毒素过敏反应1例

凡外伤、烧伤和动物咬伤等病例 ,为预防破伤风的发生 ,采用被动免疫法 ,常用破伤风抗毒素( TAT) 1 5 0 0 IU肌肉注射。 TAT临床应用机会很多 ,虽过敏反应发生率不高 ,但仍有报道发生严重过敏反应 ,甚至过敏性休克的报道 [1]。现将我们第一门诊部救治的 1例 TAT过敏反应的临床情况和救治体会报道如下。1　病例报告　　患者 ,男 ,38岁 ,因头皮外伤来门诊部就诊 ,外科清洁消毒包扎后 ,来治疗室进行 TAT治疗 ,皮试按护理操作常规 TAT 1 :1 5 0 0 IU取上液0 .1 ml加注射用水至 1 ml。以前臂内侧皮内注射0 .0 2 ml,观察 30 min,患者皮丘直…     

破伤风抗毒素精制效果的比较研究

作者对破伤风抗毒素精制效果进行了比较研究。结果表明,采用改进方法精制的3批制品F(ab′)2含量平均达到79％,而IgG含量则降为无检出;3批制品的比活性与常规法相比平均提高14％。     

破伤风抗毒素渗透压的质量控制标准

目的：通过对不同厂家破伤风抗毒素产品的渗透压浓度的调查,了解国内该产品的渗透压浓度的波动范围,为生产过程中渗透压浓度质量控制提供依据。方法采用Advanced 3250型冰点渗透压仪检测破伤风抗毒素的渗透压浓度。结果不同厂家破伤风抗毒素制品的渗透压浓度均不低于240 mOsmol/L。结论破伤风抗毒素渗透压浓度波动范围均与国外同类制品基本一致,符合欧洲药典规定。     

顶空气相色谱法检测破伤风抗毒素中的甲苯残留量

目的建立检测破伤风抗毒素中甲苯残留量的顶空气相色谱法。方法参照《中国药典》三部(2015版)"残留溶剂测定法",采用Agilent DB-1301毛细管色谱柱(15.0 m×530μm×1.00μm)及氢火焰离子化检测器;顶空进样器各参数分别为:平衡温度70℃,平衡时间30 min,定量环温度80℃,传输线温度90℃,进样量1 m L;气相色谱仪各参数为:进样室温度100℃,柱温箱温度60℃,检测器温度220℃,载气氮气流速5 m L/min,采集时间5min;并对该方法的系统适用性、专属性、线性范围、准确度和精密度、检测限和定量限进行验证及初步应用。结果甲苯保留时间2.429 min,峰面积的RSD为1.9%,以甲苯色谱峰计算得到的理论塔板数N=12 900,甲苯色谱峰与其相邻色谱峰的分离度R=6.03。方法的专属性强,制品中的其他物质不干扰甲苯的出峰;标准曲线的范围为1.0×10-4%~2.0×10-3%,相关系数r大于0.99;加标试验的回收率分别为96.0%~102.9%、100.7%~111.0%,重复性和日间精密度RSD均小于5%。检出限为2.6×10-6%,定量限为1.0×10-5%。测定10批破伤风抗毒素成品和10批破伤风抗毒素原液,其甲苯残留量均低于检出限,远低于规定的限度0.089%。结论该方法简便、快速、准确度、灵敏度高、精密度好,适用于破伤风抗毒素类制品中甲苯残留量的检测。     

应用双抗原ELISA夹心法检测破伤风抗毒素效力的试验

作者建立了一套适用于破伤风抗毒素效力检测的双抗原夹心ELISA法,对破伤风类毒素免疫后的豚鼠和马匹分别进行了血清效价测定,并和传统的小鼠攻毒法的效力检测进行了比较,结果表明：该检测方法与小鼠攻毒法对不同水平的破伤风抗毒素效力检测,结果是一致的,有很好的相关性;此法省时、特异、灵敏,可成为另一种检测破伤风抗毒素的有效方法。     

有机溶剂/去污剂病毒灭活处理对破伤风抗毒素质量的影响

目的 探索用有机溶剂/去污剂(solvent/detergent, S/D)病毒灭活处理对破伤风抗毒素质量的影响。方法 3批破伤风抗毒素样品,每批样品取3等份,向其中2份中分别加入磷酸三丁酯(tri- n -butylphosphate, TNBP)和吐温-80(Tween 80)至终质量分数为0.3%和1%;一份放置在(25±1)℃水浴中振摇6 h后取样,另一份放置在(30±1)℃水浴中振摇4 h后取样;向第3份破伤风抗毒素原液中加入等量的生理盐水混匀后室温放置作为对照。各样品经超滤后检测其效价和蛋白质含量,同时用SDS-PAGE电泳和凝胶过滤色谱柱测定。结果 破伤风抗毒素原液样品经过S/D病毒灭活处理后,其动物效价与对照相比差异无统计学意义( P >0.05),蛋白质含量、分子大小分布无明显变化,多聚体、二聚体及F(ab′) 2含量也无明显变化。结论 S/D病毒灭活处理对破伤风抗毒素的效价,蛋白质含量,分子大小分布,多聚体、二聚体及F(ab′) 2含量均无明显影响,可以作为破伤风抗毒素病毒灭活的候选方法。     

人破伤风免疫球蛋白及其应用

临床上对破伤风的预防与治疗通常使用破伤风抗毒素(TAT)和人破伤风免疫球蛋白(TIG),前者由于来源于动物血清,常常发生过敏反应和血清病,而后者的过敏反应率很低,预防效果好。本文就TAT和TIG的应用效果比较、TIG的剂量、生产开发、市场前景等作了全面的综述。     

抗破伤风毒素人源单克隆抗体中和效力的研究

对抗破伤风毒素人源单克隆抗体Ｇ２、Ｇ６、Ｇ２＋Ｇ６与精制破伤风抗毒素进行了中和效力比较试验。结果表明,Ｇ２、Ｇ６二株单抗的中和毒力活性均比精制破伤风抗毒素的高。Ｇ２和Ｇ６混合后呈完全中和。表明这些人源单抗在中和毒力方面能够取代精制破伤风抗毒素,可满足临床试验需求。     

新型佐剂在破伤风抗毒素血浆生产中的应用研究

应用两种新型佐剂(Montan inde ISA 50V和Montan inde ISA 206)分别乳化精制破伤风类毒素制备成油乳抗原,免疫豚鼠三程,并与弗氏不完全佐剂(FIA)进行对比;在弗氏不完全佐剂免疫马匹到第7程后,改换新型佐剂抗原免疫三程(第8、9、10程)。采用小鼠中和试验测定血清效价。结果表明,两种新型佐剂使用安全,稳定性良好,且操作方便;使用Montan inde ISA 50V免疫豚鼠与马匹的免疫效果均有明显优势,Montan inde ISA 206次之,FIA效果相对较差。表明新型佐剂Montan inde ISA 50V的应用前景良好。     

Quantitation of commercial equine tetanus antitoxin by competitive enzyme-linked immunosorbent assay

In the USA, the potency of commercially prepared equine tetanus antitoxin is determined by the method outlined in the Code of Federal Regulations, Title 9, Part 113.451. In the current test, commercial equine tetanus antitoxin is tested by a toxin neutralization test in guinea pigs. The in vivo test measures antitoxin content through effectiveness of protection of guinea pigs injected with diluted mixtures of antitoxin and a standard toxin. A competitive enzyme-linked immunosorbent assay, designed as an in vitro alternative to the in vivo test, measures antitoxin content based on a competitive reaction between standard or unknown serum and murine monoclonal antibody specific for tetanus toxin. The monoclonal antibody used in the assay delayed death in mouse passive protection studies and reacted with the C fragment of tetanus toxin. No cross-reaction was observed when the antibody was tested with the toxins of Clostridium chauvoei, C. novyi, C. perfringens, or C. sordellii. The in vitro test will measure the antitoxin content of serum samples containing 100-1500 units of antitoxin. Tetanus antitoxin titers obtained by the competitive enzyme-linked immunosorbent assay compared favorably with the toxin neutralization test conducted in guinea pigs. The in vitro assay serves as a feasible alternative to the in vivo test because it can be completed in less time, is reproducible, and eliminates the use of test animals.     

In vitro titration of tetanus antitoxin

In vitro methods are used as an alternative to the expensive and time-consuming official method in mice for the titration of tetanus antibodies. Numerous techniques have been developed and used for this purpose, but with mixed results. The difficulty is to get good correlation with in vivo units whatever the vaccinal status of the individual. Today, some of these serological techniques have been virtually abandoned in favor of others which are becoming widespread. RIA is still used, but only in laboratories which already have the necessary equipment and skill. It is gradually replaced by ELISA, a rather delicate technique which however has kept its promises and is amenable to further progress. Passive haemagglutination has been greatly improved by the use of turkey erythrocytes. With this modification, this technique becomes reliable and sensitive and is apparently able to give good results for a large number of sera in case of mass investigation as well as for a quick estimate of the protective level in an individual.