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题名 酶联免疫S-HpAg快速诊断试剂盒的制备
姓名 李静贤
院系 生命科学学院
专业 应用生物化学
学位名称 理学硕士
外文题名 Rapid Detection of S-HpAg by ELISA
第一导师姓名 李红玉
关键词 幽门螺杆菌;可溶性Hp抗原; 杂交瘤;单克隆抗体;酶联免疫吸附
外文关键词 Helicobacter pylori (H. pylori);Soluble H. pylori antigen;hybridoma;Monoclonal Antibodies (McAbs);enzyme linked immunosorbent assay(ELISA)
学科 理学
摘要 1982年澳大利亚学者Warran和Marshall从慢性胃炎患者胃粘膜中培养出幽门螺杆菌(Helicobacter pylori ,H. pylori)以来,H. pylori受到了极大的关注。H. pylori是一种传染率高的有害菌,全世界约有50%人口被其感染。 H. pylori与胃癌(gastric carcinoma,GC)及胃MALT(Mucosa Association Lymphoid Tissue)恶性淋巴瘤的发生关系密切。此外,H. pylori 还可引起其它一些疾病,如冠心病,儿童生长迟缓或消化不良等。WHO和国际抗癌联盟已将其列为1类致癌因子。 目前,诊断幽门螺杆菌感染的方法从检测手段上分为侵入性和非侵入性两大类。侵入性诊断方法最大的缺点,或者说受到的限制是必需通过胃镜取样本,造成胃病患者额外增加一些损伤与痛苦。非侵入性诊断方法也有缺陷,如放射性同位素本身会污染环境,需用气体同位素质谱仪测定,以及受抗体回忆反应影响造成假阴(阳)性结果。 Hp-SAg 检测作为一种新的Hp检测方法,具有检测快速、经济、准确、宜于大规模普查等优点,且不受抗体回忆反应影响,结果稳定、特异性高,可用于Hp的临床诊断、根除疗效观察和随访复查等,也适合于儿童及孕妇检测。 本实验建立了4株稳定分泌Hp单克隆抗体的细胞株:E4、B7、G2、G3.完成了单克隆抗体的筛选、制备、纯化、鉴定、工作单抗配对等方面的工作,为进一步进行临床试验打下了基础。同时,Hp的酶联免疫试剂盒还为胶体金试剂盒的研制作出了重要的前期工作,给后者打下了必要的基础。
外文摘要 Helicobacter pylori (H. pylori) has been attracted more attention since it was separated from gastric mucosa and culture successfully by Warran and Marshall in 1982. . H. pylori is a worldwide epidemic bacteria. there are about half people infect H. pylori H. pylori is close with peptic ulcer.. it has near relationship between H. pylori and gastric carcinoma (or Mucosa Association Lymphoid Tissue Lymphoma ) . Hp can also give rise to eight other diseases: coronary heart disease colic, growth retardation and dyspepsia of childhood and so on. It was list as Grade 1 Carcinogen. Recently, there are two main ways, invade and non-invade, to diagnose the infection of Hp. But the invade way needs to have a gastroscope to got living gastric mucous membrane specimen that will bring the patient many unnecessary pains. At the same time, the non-invade way has its limitation too, such as using radioactive isotope is which will pollute environment, needing special facility, and being affected by memorizing reaction of serum antibody then bring false negative or false positive result The detection of Hp soluble antigen could be used as a new method which is accurate, convenient, not being affected by memorizing reaction of serum antibody, more sensitive, more specific and suitable for clinical diagnosis, evaluation of eradication and follow-up of Hp as well as for detection in children and pregnant women Four cell strains which can steadily excrete McAbs(monoclonal antibodies) were established in this study, named E4、B7、G2、G3. The filtration, preparation, purification, identify of the McAbs and work partnership which are the foundation of the clinical trial were finished. At the mean time The Hp-ELISA detection kit is the necessary foundation of the study of golden colloid kit.
研究领域 应用生物化学
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