兔源抗钩端螺旋体—多克隆抗体的制备及治疗效果研究

发布时间：2018-04-09 00:08

本文选题：钩端螺旋体病　切入点：多克隆抗体　出处：《吉林大学》2017年硕士论文

【摘要】：钩端螺旋体病,是由钩端螺旋体引起的一种最重要的被忽视热带人兽共患病之一,已经对人类健康产生了重要威胁。某些病原性疾病,如钩端螺旋体病,缺乏有效的治疗方法,而多克隆抗体治疗方法是一种极具潜力的治疗方法。首先经过改良后科学的免疫程序,用黄疸出血群的赖型56601株钩体免疫接种新西兰大白兔后,使耐受动物兔子血清内产生了抗钩端螺旋体的抗体,显微凝集实验鉴定了该免疫血清内具有与钩端螺旋体结合的抗体。经过正辛酸-硫酸铵两步沉淀法纯化后,获得多克隆抗体。用56601钩体与秋季群秋季型56606钩体分别作为抗原,经过ELISA检测,该多克隆抗体被两种血清型的钩体所识别。本研究评估了兔源多克隆抗体针对同血清型(赖型)钩端螺旋体感染或一种异血清型(秋季型)钩端螺旋体感染的早期治疗效果。该多克隆抗体提高了存活率,无论56601钩体感染还是56606钩体感染。在同血清型钩端螺旋体病早期治疗中提高了金黄地鼠钩体病的存活率,而且呈剂量依赖性,因此也为其他动物或人的钩端螺旋体病提供了剂量参考;在异血清型钩端螺旋体病早期治疗中,高剂量组也显著提高了存活率,虽然并未做到完全保护。为以后研究制备抗多血清型多克隆抗体奠定了基础。通过病理组织学变化的观察,治疗组动物的肝脏,肾脏和肺脏的病例损伤明显减轻,说明该多克隆抗体具有病理上的治疗作用。而且通过qPCR证明该多克隆抗体具有清除器官内钩端螺旋体的作用。另外还进行了晚期治疗试验,多克隆抗体和抗生素的治疗之间并未出现显著差异,但发现该多克隆抗体和抗生素联合用药,相比其他组可显著提高存活率。本研究认为该兔源多克隆抗体在早期治疗中对同血清型和一种异血清型的钩端螺旋体感染的金黄地鼠具有保护效果,并且在晚期治疗中和抗生素的联合用药可以显著提高存活率。
[Abstract]:Leptospirosis, one of the most important neglected zoonosis caused by leptospirosis, has posed an important threat to human health.Some pathogenic diseases, such as leptospirosis, lack of effective treatment, and polyclonal antibody therapy is a promising treatment.After inoculating New Zealand white rabbits with 56601 Leptospira strains of jaundice haemorrhage group, the antibody against leptospirosis was produced in the sera of tolerant rabbits.The antibody binding to leptospira was identified by microagglutination assay.The polyclonal antibody was purified by two-step precipitation method of n-octanoic acid-ammonium sulfate.Leptospira 56601 and fall Leptospira 56606 were used as antigens respectively. The polyclonal antibody was identified by two serotypes of Leptospira after ELISA detection.The aim of this study was to evaluate the early therapeutic effects of rabbit polyclonal antibodies against leptospirosis infection of the same serotype (Leptospira) or an alloserotype (autumn type) infection of Leptospira leptospira.The polyclonal antibody improves survival rate, whether 56601 leptospirosis or 56606 leptospirosis.In the early treatment of leptospirosis, the survival rate of hamster leptospirosis was increased in a dose-dependent manner, which provided a dose reference for other animals or human leptospirosis.In the early treatment of leptospirosis, the survival rate was also significantly increased in the high dose group, although not fully protected.It lays a foundation for the preparation of polyclonal antibodies against polyserotypes in the future.Through the observation of histopathological changes, the damage of liver, kidney and lung in the treatment group was obviously alleviated, which indicated that the polyclonal antibody had pathological therapeutic effect.Moreover, the polyclonal antibody was proved to be capable of scavenging leptospirosis in organs by qPCR.There was no significant difference between the treatment of polyclonal antibody and antibiotics, but it was found that the combination of polyclonal antibody and antibiotics could significantly improve the survival rate.This study suggested that the rabbit polyclonal antibody had protective effect on hamsters infected with leptospira leptospira in the early stage of treatment.The combination of antibiotics and advanced treatment can significantly improve survival rate.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S855

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