旋毛虫排泄分泌物抗原在鼠和猪免疫学诊断中的应用研究

发布时间：2018-06-11 10:01

本文选题：旋毛虫 + 排泄分泌物　；参考：《吉林大学》2015年硕士论文

【摘要】：旋毛虫病是一种重要的食源性人畜共患寄生虫病，呈世界性分布，由组织栖线形旋毛虫引起。人类通过生食或半生食含旋毛虫肌幼虫的肉类（主要是猪肉）而感染旋毛虫病。此病目前在一些发展中国家（如泰国、阿根廷及中国等）仍常有暴发，究其原因除了缺少有效的预防控制措施外，关键在于至今没有良好的诊断方法。旋毛虫病的临床症状表现一般不具明显特征，且易与其它传染病相混淆，因此临床诊断较为困难。虽然肌肉活检发现幼虫或包囊即可确诊，但在轻度感染早期往往不易检出，即使是感染后期，因受组织局限性影响，活检阳性率也仅有50%左右。为此，寻找一种敏感性高、特异性强的检测诊断方法对于旋毛虫病的诊、防、治具有十分重要的意义，其关键是筛选到有效的免疫学检测诊断抗原。本研究拟以小鼠为实验动物模型，并应用于临床出栏猪的诊断研究，利用中国河南猪旋毛虫T1分离株(ISS534)分别感染小鼠和猪，采用旋毛虫不同发育时期的排泄分泌物抗原（ES）作为诊断抗原，应用ELISA方法检测感染小鼠和猪60天内不同天数的血清抗旋毛虫IgM和IgG抗体水平，并采用Excel软件绘制其抗体消长规律曲线并进行数据分析。结果表明：对于小鼠模型，，三种抗原检测到抗旋毛虫抗体IgM均成“波峰波谷”形式的变化曲线，可以利用旋毛虫三个发育时期ES抗原的混合物来检测旋毛虫45天前的感染（即5-14天的感染检测IgM、15-45天检测IgG），感染45天之后可以利用单一肌幼虫时期ES抗原检测IgG，最有意义的发现是利用旋毛虫成虫期和新生幼虫期的ES抗原可以有效检测旋毛虫的早期感染；对于猪的应用研究，低剂量感染最好利用旋毛虫不同发育时期的3至5种分泌物抗原的混合物来检测旋毛虫2个月前的感染；中高剂量感染可利用单一感染后10小时肠道期肌幼虫ES抗原来检测感染60天内血清抗旋毛虫IgM抗体。本研究成功获悉旋毛虫感染宿主后不同发育时期刺激机体产生抗旋毛虫抗体的消长规律，确定了旋毛虫不同发育时期的免疫学诊断抗原，为进一步合理利用旋毛虫不同发育时期的排泄分泌物抗原建立起有效检测旋毛虫感染的诊断制剂的研制提供了重要的理论基础和科学依据。
[Abstract]:Trichinellosis is an important zoonotic parasitic disease caused by histopathic nematode. Humans are infected with trichinellosis by eating raw or semi-raw meat (mainly pork) containing the muscle larvae of Trichinella spiralis. There are still many outbreaks in some developing countries (such as Thailand, Argentina and China). Besides the lack of effective prevention and control measures, the key lies in the lack of good diagnostic methods. The clinical symptoms of trichinellosis are not obvious and easily confused with other infectious diseases, so clinical diagnosis is difficult. Although the larva or cyst can be diagnosed by muscle biopsy, it is not easy to detect in the early stage of mild infection. Even in the late stage of infection, the positive rate of biopsy is only about 50% due to the influence of tissue localization. Therefore, it is of great significance for the diagnosis, prevention and treatment of trichinellosis to find a highly sensitive and specific method for detection and diagnosis. The key is to screen effective immunological diagnostic antigens. This study was designed to use mice as an experimental animal model and to be used in the clinical diagnosis of pigs. The mice and pigs were infected with ISS534, a T1 strain of Trichinella spiralis from Henan Province, China, respectively. The excretory secretion antigens of Trichinella spiralis were used as diagnostic antigens. Elisa was used to detect the levels of anti-trichinella IgM and IgG in sera of infected mice and pigs for different days within 60 days. Excel software was used to draw the curve of antibody growth and decline and to analyze the data. The results showed that for the mouse model, the three antigens detected the change curve of anti-trichinella antibody IgM in the form of "peak and trough". The mixture of es antigens of three developmental stages of Trichinella spiralis can be used to detect the infection of Trichinella spiralis 45 days ago (i.e. 5-14 days of infection detection of IgMN 15-45 days of detection of IgGN). After 45 days of infection, the single muscle larvae period es antigen can be used to detect IgGs. It is significant to find that the early infection of Trichinella spiralis can be detected effectively by using es antigens of adult and newborn larvae. For the application study of pigs, low dose infection is best to detect trichinella infection 2 months ago by using a mixture of 3 to 5 secretions antigens of Trichinella spiralis at different developmental stages. The serum anti-trichinella IgM antibody could be detected by using the es antigen of the intestinal muscle larvae 10 hours after a single infection in the middle and high dose of infection. In this study, we have learned the law of the growth and decline of anti-trichinella antibody in different development period after Trichinella spiralis infection, and determined the immunological diagnosis antigen of Trichinella spiralis in different development period. It provides an important theoretical basis and scientific basis for the development of an effective diagnostic preparation for the detection of Trichinella spiralis infection by using the excretory secretion antigens of Trichinella spiralis at different developmental stages.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.7

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