首页 | 本学科首页   官方微博 | 高级检索  
     

抗嗜肺军团菌血清8型单克隆抗体的制备及双抗体夹心酶联免疫吸附试验检测法的建立
引用本文:郭景玉,朱紫雯,王津,杨瑞馥,宋亚军. 抗嗜肺军团菌血清8型单克隆抗体的制备及双抗体夹心酶联免疫吸附试验检测法的建立[J]. 生物技术通讯, 2009, 20(4): 488-490,519. DOI: 10.3969/j.issn.1009-0002.2009.04.010
作者姓名:郭景玉  朱紫雯  王津  杨瑞馥  宋亚军
作者单位:军事医学科学院,微生物流行病研究所,病原微生物生物安全国家重点实验室,北京,100071
摘    要:目的:制备针对嗜肺军团茵血清8型的单克隆抗体,并建立双抗体夹心酶联免疫吸附试验(ELISA)检测方法。方法:用甲醛灭活的嗜肺军团菌血清8型菌免疫BALB/c小鼠,采用杂交瘤技术制备抗嗜肺军团菌血清8型单克隆抗体,建立双抗夹心ELISA检测方法。结果:研制出8株能特异性分泌抗嗜肺军团菌血清8型单克隆抗体的杂交瘤细胞株,Ig类型分别为IgM(2株)、IgG,(1株)和IgG,(5株);利用IgG1型单抗6G10与6C7配对,建立了双抗夹心ELISA检测方法,该方法的最低检出浓度为2.6×10^5cfu/mL,除与金黄色葡萄球菌有微弱的交叉反应外,与14株其他血清型嗜肺军团菌、17株非嗜肺军团菌及11株非军团菌均无交叉反应,具有较高的特异性。结论:制备了具有高特异性和亲和力的抗嗜肺军团菌血清8型单克隆抗体,并建立了双抗夹心EUSA检测方法。

关 键 词:嗜肺军团菌血清8型  单克隆抗体  双抗夹心酶联免疫吸附试验

Preparation of Monoclonal Antibodies Against Legionella pneumophila Serogroup 8 and Establishment of Antibody-Sandwich ELISA Method for Detecting this Pathogen
GUO Jing-Yu,ZHU Zi-Wen,WANG Jin,YANG Rui-Fu,SONG Ya-Jun. Preparation of Monoclonal Antibodies Against Legionella pneumophila Serogroup 8 and Establishment of Antibody-Sandwich ELISA Method for Detecting this Pathogen[J]. Letters in Biotechnology, 2009, 20(4): 488-490,519. DOI: 10.3969/j.issn.1009-0002.2009.04.010
Authors:GUO Jing-Yu  ZHU Zi-Wen  WANG Jin  YANG Rui-Fu  SONG Ya-Jun
Affiliation:(Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China)
Abstract:Objective: To preparate monoclonal antibodies(McAbs) anginst Legionella pneumophila serogroup 8 and establish an antibody-sandwich ELISA method for detection of the organism. Methods: BALB/c mice were immunized by formalin-killed L.pneumophila serogroup 8 and McAbs were prepared by using hybridoma technique. By using two McAbs, an antibody-sandwich ELISA was established. Results: Eight hybridomas producing antibodies against L.pneumophila serogroup 8 were obtained. Ig isotypes of the McAbs were IgM, IgG3 and IgG1. An antibody-sandwich ELISA was established and the sensitivity for sonicated cells was about 2.6x105 cfu/mL. No cross-reactivity was detectable in 14 other serogroup L.pneumophila, 17 non-pneumophila legionellae and 11 other bacteria, except Staphylococcus aureus which has a weak cross reaction. Conclusion: Two hybridomas producing high specificity and affinity monoclonal antibodies against L. pneumophila serogroup 8 can provide for antibody-sandwich ELISA.
Keywords:Legionella pneumophila serogroup 8  monoclonal antibody  antibody-sandwich ELISA
本文献已被 维普 万方数据 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号