酶联免疫S-HpAg快速诊断试剂盒的制备
发布时间：2018-12-13 00:29
【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。 S—HpAg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株稳定分泌抗人HP单克隆抗体细胞株: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。已申请一项国家发明专利,申请号为2005102002286。
[Abstract]:Since Australian scholars Warran and Marshall cultured Helicobacter pylori (Helicobacter pylori,H.pylori) from gastric mucosa of patients with chronic gastritis in 1982, H.pylori has received great attention. H.pylori is a harmful bacterium with high infection rate. About 50% of the world's population is infected with H.pylori, which is closely related to the occurrence of gastric cancer (gastric carcinoma,GC) and gastric MALT (Mucosa Association Lymphoid Tissue) malignant lymphoma. In addition, H.pylori can also cause other diseases, such as coronary heart disease, growth retardation in children or indigestion, etc. WHO and the International Union against Cancer have classified it as a carcinogen of class 1. At present, the diagnostic methods of Helicobacter pylori infection can be divided into invasive and non invasive methods. The greatest drawback of invasive diagnosis, or limitation, is the need for gastroscopy to cause additional damage and pain in patients with gastropathy. Noninvasive diagnostic methods are also flawed, such as radioisotopes themselves that pollute the environment, gas isotope mass spectrometers and pseudo (positive) results due to the effect of antibody recall reactions. As a new method of HP detection, S-HpAg detection has the advantages of rapid, economical, accurate, suitable for large-scale screening, and can be used in the clinical diagnosis of HP without the influence of antibody recall reaction. The observation of eradication effect and follow-up examination are also suitable for children and pregnant women. In this study, we established four stable secreting monoclonal antibody cell lines against human HP: E4B7G2G3. The selection, preparation, purification, identification and matching of monoclonal antibodies were completed, which laid a foundation for further clinical trials. At the same time, the enzyme linked immunosorbent kit (Elisa) of HP has also made an important preliminary work for the development of colloidal gold kit and laid a necessary foundation for the latter. Has applied for a national invention patent, the application number is 200510 2002286.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
[Abstract]:Since Australian scholars Warran and Marshall cultured Helicobacter pylori (Helicobacter pylori,H.pylori) from gastric mucosa of patients with chronic gastritis in 1982, H.pylori has received great attention. H.pylori is a harmful bacterium with high infection rate. About 50% of the world's population is infected with H.pylori, which is closely related to the occurrence of gastric cancer (gastric carcinoma,GC) and gastric MALT (Mucosa Association Lymphoid Tissue) malignant lymphoma. In addition, H.pylori can also cause other diseases, such as coronary heart disease, growth retardation in children or indigestion, etc. WHO and the International Union against Cancer have classified it as a carcinogen of class 1. At present, the diagnostic methods of Helicobacter pylori infection can be divided into invasive and non invasive methods. The greatest drawback of invasive diagnosis, or limitation, is the need for gastroscopy to cause additional damage and pain in patients with gastropathy. Noninvasive diagnostic methods are also flawed, such as radioisotopes themselves that pollute the environment, gas isotope mass spectrometers and pseudo (positive) results due to the effect of antibody recall reactions. As a new method of HP detection, S-HpAg detection has the advantages of rapid, economical, accurate, suitable for large-scale screening, and can be used in the clinical diagnosis of HP without the influence of antibody recall reaction. The observation of eradication effect and follow-up examination are also suitable for children and pregnant women. In this study, we established four stable secreting monoclonal antibody cell lines against human HP: E4B7G2G3. The selection, preparation, purification, identification and matching of monoclonal antibodies were completed, which laid a foundation for further clinical trials. At the same time, the enzyme linked immunosorbent kit (Elisa) of HP has also made an important preliminary work for the development of colloidal gold kit and laid a necessary foundation for the latter. Has applied for a national invention patent, the application number is 200510 2002286.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
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