布鲁氏菌

发布时间：2018-12-29 17:28

【摘要】：布鲁氏菌病严重危害着人类的健康和畜牧业的快速发展,它是一种动物源性、自然疫源性人兽共患病。莱姆病是一种人兽共患传染性疾病,是由蜱传播的若干个不同基因种的伯氏疏螺旋体引起的疾病。目的:(1)本研究基于血清学诊断方法,建立一种快速、便捷的适用于基层诊断布鲁氏菌的免疫胶体金试纸条检测方法。(2)本研究应用免疫渗滤技术,建立一种早期鉴别莱姆病的快速诊断方法。方法:(1)提取纯化羊种布鲁氏菌强毒株16M LPS,对其纯度、浓度进行鉴定后,以LPS作为包被抗原,优化完善胶体金免疫层析技术以及试纸条的制作条件,完成试纸条的组装后,对不同动物、人类血清样品进行检测,并与虎红平板凝集试验、试管凝集试验结果进行对比,分析以16M LPS作为包被抗原的试纸条的可行性。(2)使用石河子大学人兽共患病实验室构建的莱姆重组蛋白Bmp A,经过纯化、浓缩,达到最佳包被浓度后,优化免疫渗滤技术的条件,使用免疫渗滤技术对疑似莱姆患者血清及未知绵羊血清进行检测,通过检测结果,探究莱姆膜脂蛋白Bmp A作为包被抗原应用免疫渗滤技术,对莱姆病诊断的可行性。结果:(1)免疫胶体金试纸条LPS最佳包被浓度为2.0 mg/m L。440份血清样品检测结果为:试纸条与试管凝集试验的符合率为95.45%,明显高于虎红平板凝集试验与试管凝集试验的符合率(79.55%);且试纸条的阳性检出率为84.32%,试管凝集试验阳性检出率为86.59%,相差较小。(2)重组蛋白Bmp A以2.0 mg/m L为制作免疫渗滤卡盒的最适包被量,用免疫渗滤卡对8份确定感染莱姆病患者的血清进行检测,检测出3份阳性血清;对30份的绵羊血清进行检测,检测出2份阳性血清,经过WB验证结果一致。结论:本研究建立了以LPS作为包被抗原的布鲁氏菌病胶体金试纸条诊断方法,可用于基层对布鲁氏菌病的初步筛查;优化免疫渗滤技术,建立了以纯化的Bmp A莱姆病伯氏疏螺旋体膜脂蛋白作为包被抗原的莱姆病免疫渗滤卡诊断方法。为布鲁氏菌病和莱姆病的临床诊断提供了技术支持。
[Abstract]:Brucellosis seriously endangers human health and the rapid development of animal husbandry, it is an animal-derived, natural epidemic zoonosis. Lyme disease is a zoonotic infectious disease caused by several tick-borne spirochetes. Objective: (1) based on the method of serological diagnosis, this study established a rapid and convenient method for the detection of immuno-colloidal gold strip for the diagnosis of brucella. To establish a rapid diagnostic method for early differential diagnosis of Lyme disease. Methods: (1) the purity and concentration of 16m LPS, were extracted and purified from Brucella virulent strain of sheep. LPS was used as the coating antigen to optimize the preparation conditions of colloidal gold immunochromatography and test strip. After the assembly of the test strip, different animal and human serum samples were detected, and the results were compared with the results of the Tiger Red plate agglutination test and the test tube agglutination test. The feasibility of using 16m LPS as the test strip of coated antigen was analyzed. (2) the recombinant protein Bmp A was constructed by the laboratory of zoonoses of Shihezi University. After purification and concentration, the recombinant protein Bmp A was obtained to the best coating concentration. The condition of immune filtration technique was optimized, and the serum of suspected Lyme patients and unknown sheep serum were detected by using immunofiltration technique. Through the test results, the application of immunofiltration technique was studied as the coating antigen of Lim membrane lipoprotein Bmp A. The feasibility of diagnosis of Lyme disease. Results: (1) the best coating concentration of LPS was 2.0 mg/m L. 440 serum samples. The results showed that the coincidence rate between the test strip and the test tube agglutination test was 95.45%. The coincidence rate of Tiger red plate agglutination test and test tube agglutination test (79.55%) was higher than that of Tiger red plate agglutination test (79.55%). The positive rate of the test strip was 84.32 and the positive rate of the test tube agglutination test was 86.59, the difference was small. (2) the recombinant protein Bmp A used 2.0 mg/m L as the optimal envelope for making the immune leachate card box. Eight sera from patients with Lyme disease were detected with immune leachate and 3 positive sera were detected. Two positive sera were detected from 30 sheep sera, and the results were consistent by WB. Conclusion: in this study, a method for the diagnosis of brucellosis colloidal gold strip with LPS as the coating antigen was established, which can be used for the primary screening of brucellosis at the basic level. A method for the diagnosis of Bmp A Lyme disease by using purified membrane lipoprotein of Borrelia burgdorferi as antigen was established by optimizing immunofiltration technique. It provides technical support for the clinical diagnosis of brucellosis and Lyme disease.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S855

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