本文關(guān)鍵詞： 甲型副傷寒沙門氏菌 16S rRNA 外膜蛋白 單克隆抗體　出處：《中國藥品生物制品檢定所》2006年碩士論文　論文類型：學(xué)位論文

【摘要】：近年來，我國南方一些地區(qū)以及東南亞某些國家均有甲型副傷寒沙門氏菌流行的報道。甲型副傷寒暴發(fā)流行，顯示了傷寒流行菌型變遷的新特點，即由原來以傷寒沙門氏菌流行為主轉(zhuǎn)為以甲型副傷寒沙門氏菌為主導(dǎo)菌，因此對甲型副傷寒沙門氏菌的研究非常有必要。本論文從兩個方面對甲型副傷寒沙門氏菌進(jìn)行了研究，即甲型副傷寒沙門氏菌外膜蛋白免疫原性的研究和甲型副傷寒沙門氏菌菌體抗原單克隆抗體的制備。 論文第一部分的主要研究目的在于：探索應(yīng)用單一或混合的甲型副傷寒沙門氏菌外膜蛋白作為保護(hù)性抗原，進(jìn)而作為甲型副傷寒的候選疫苗的可能性。首先對本室保存的6株甲型副傷寒沙門氏菌株進(jìn)行鑒定和篩選。6株菌均符合革蘭氏陰性菌的形態(tài)特點，生化反應(yīng)結(jié)果顯示6株菌均為甲型副傷寒沙門氏菌，可信度為97％，并表現(xiàn)出不發(fā)酵木糖，也不產(chǎn)生H_2S的甲型副傷寒沙門氏菌主要生化特征。血清學(xué)檢測顯示6株菌與沙門氏菌診斷血清(02、012、Ha)發(fā)生了較強(qiáng)的凝集。建立小鼠攻毒模型對6株菌的毒力進(jìn)行比較，，選擇出毒力較強(qiáng)的50973為后續(xù)實驗用菌株。借助分子生物學(xué)技術(shù)對50973的16S rDNA進(jìn)行了測序，結(jié)果與GeneBank中已知的甲型副傷寒沙門氏菌的16s rRNA基因序列進(jìn)行比對，兩者同源性達(dá)到99％；其后建立了體外殺菌抗體檢測的實驗方法，確定了最佳的菌液濃度為1000CFU／ml，反應(yīng)時間為16—18h，乳兔的補(bǔ)體作為補(bǔ)體來源等，并進(jìn)行了驗證；最后建立了甲型副傷寒沙門氏菌外膜蛋白的提取方法，在此基礎(chǔ)上應(yīng)用SDS—PAGE電泳來分離外膜蛋白的各個組分，并用電洗
[Abstract]:In recent years, some areas in southern China and some countries in Southeast Asia have reported the epidemic of Salmonella paratyphoid A. the outbreak of paratyphoid A shows the new characteristics of the change of epidemic type of typhoid. That is, from the original epidemic of Salmonella typhimurium to the predominant bacillus paratyphoid A. Therefore, it is very necessary to study the salmonella paratyphoid A. in this paper, we studied the salmonella paratyphoid A from two aspects. The immunogenicity of the outer membrane protein of Salmonella paratyphi A and the preparation of monoclonal antibody against Salmonella paratyphi A. The main purpose of the first part of the thesis is to explore the application of a single or mixed outer membrane protein of Salmonella paratyphi A as a protective antigen. At first, 6 strains of Salmonella paratyphoid A preserved in our room were identified and screened. All the 6 strains were in accordance with the morphological characteristics of Gram-negative bacteria. The results of biochemical reaction showed that all of the 6 strains were paratyphoid A Salmonella A, and the reliability was 97%. No H2S production was found in the main biochemical characteristics of Salmonella paratyphi A. serological tests showed that 6 strains of bacilli and the diagnostic serum of Salmonella spp. Ha. the virulence of 6 strains of bacteria was compared by establishing a mouse model of attacking virus. The highly virulent strain 50973 was selected for further experiment. The 16s rDNA of 50973 was sequenced by molecular biology technique. Results compared with the 16s rRNA gene sequence of Salmonella paratyphoid A in GeneBank, the homology of the two genes was 99%. After that, an experimental method for the detection of bactericidal antibody in vitro was established. The optimal concentration of bactericidal liquid was 1000 CFU / ml, the reaction time was 16-18 h, and the complement of milk rabbit was used as the source of complement. And verified; Finally, the extraction method of outer membrane protein of Salmonella paratyphi A was established. On the basis of this, SDS-PAGE electrophoresis was used to separate the components of outer membrane protein, and electrowashing was carried out.
【學(xué)位授予單位】：中國藥品生物制品檢定所
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

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