重组SjP38抗原诊断日本血吸虫病的研究

发布时间：2018-01-13 02:15

本文关键词：重组SjP38抗原诊断日本血吸虫病的研究　出处：《第一军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：血吸虫病是当今最重要的公共卫生疾病之一,全世界有2亿人口被感染,其中有1.2亿患者。血吸虫病的主要危害在于虫卵所分泌的可溶性虫卵抗原(soluble egg antigen, SEA)导致的肉芽肿病变,患者在晚期时易引起肝脾肿大,门脉高压和腹水,丧失劳动力,甚至失去生命。因此,实现早期诊断,成为血吸虫病防治工作的重点。血吸虫病常用诊断方法主要有病原学和免疫学方法。传统的病原学方法有较高特异性但敏感性较差,而且多数病原学方法操作繁琐,不利于普查。相对而言,免疫学方法是血吸虫病诊断的发展方向,目前存在的主要问题是:1) 诊断用抗原、抗体的大批量来源及其质量监控问题;2) 循环抗体(CAb)检测和循环抗原(CAg)检测各有利弊,临床方法以检测IgG抗体的存在为主,但难以区分现症与既往感染;3) 早期诊断的效果不理想;4) 诊断方法的进一步标准化、简易快速化、产业化而真正面向基层与现场。 P38蛋白是SEA中分子量约为38kD的分子,大量实验证明曼氏血吸虫天然和重组P38蛋白具有良好的免疫原性,具有潜在的抗病免疫和诊断应用价值。在本研究室先前的工作中,周晓红等通过可溶性虫卵(SEA)与感染兔血清Western-blot实验,发现在38kD左右蛋白能识别4周血清;表明该蛋白参与早期免疫反应。由于天然SjP38蛋白来源有限,且纯化过程繁琐。本室已经克隆了日本血吸虫主要虫卵P38基因(SjP38)的编码序列,本研究则进一步深入研究重组SjP38蛋白(rSjP38)的表达、纯化和rSjP38单克隆抗体的制备及其在血吸虫病诊断中的应用。
[Abstract]:Schistosomiasis is one of the most important public health diseases. 200 million people worldwide are infected with schistosomiasis. Among them, 120 million patients. The main harm of schistosomiasis is the granulomatous lesion caused by soluble egg antigen-secreted by eggs. It is easy to cause hepatosplenomegaly, portal hypertension and ascites, loss of labor force and even loss of life in patients with advanced stage. Therefore, early diagnosis has become the focus of schistosomiasis prevention and control. The common diagnostic methods of schistosomiasis include etiology and immunology. The traditional etiological methods have high specificity but poor sensitivity, and most of the etiological methods are complicated to operate. Relatively speaking, immunological method is the development direction of schistosomiasis diagnosis. The main problems at present are: 1) Antigen for diagnosis, mass source of antibody and its quality control; 2) the detection of circulating antibody and circulating antigen (CAG) have their advantages and disadvantages. The main clinical method is to detect the presence of IgG antibody, but it is difficult to distinguish the present disease from the previous infection. 3) the effect of early diagnosis is not ideal; 4) the further standardization of diagnostic methods, simple and rapid, industrialization and truly oriented to the grass roots and the scene. P38 protein is a molecular weight of about 38 KD in SEA. A large number of experiments have proved that the natural and recombinant P38 protein of Schistosoma mansoni has good immunogenicity. In the previous work of our laboratory, Zhou Xiaohong and so on through the soluble egg sea) and the infection rabbit serum Western-blot experiment. It was found that the protein of 38 KD could recognize the serum for 4 weeks. It is suggested that this protein is involved in the early immune response due to the limited source of natural SjP38 protein. The P38 gene of Schistosoma japonicum has been cloned and the coding sequence of SjP38) has been cloned. In this study, the expression and purification of recombinant SjP38 protein rSjP38, the preparation of monoclonal antibody against rSjP38 and its application in diagnosis of schistosomiasis were further studied.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

【引证文献】