Sithigorngul W, Rengpipat S, Tansirisittikul A, Rukpratanporn S, Longyant S, Chaivisuthangkura P, Sithigorngul P
Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok, Thailand.
Lett Appl Microbiol. 2006 Oct;43(4):436-42. doi: 10.1111/j.1472-765X.2006.01969.x.
The present study was aimed to produce monoclonal antibodies (MAbs) for simple and specific identification of Vibrio alginolyticus infection in shrimp.
Mice were immunized with heat killed V. alginolyticus four times at 2-week intervals. The best response mouse was used for spleen donor in hybridoma production. Screening of hybridoma clones producing desired antibodies was performed by dot blotting against V. alginolyticus and other bacterial species, Western blotting and immunohistochemistry of infected shrimp tissues. Four groups of MAbs were obtained; the first group of MAbs demonstrated their limited specificity only to V. alginolyticus used for immunization, while the second and the third groups recognized all three isolates of V. alginolyticus used for testing. The fourth group of MAbs bound to all three isolates of V. alginolyticus and also recognized Vibrio parahaemolyticus, Vibrio harveyi, Vibrio fluvialis and Vibrio vulnificus but did not bind to Vibrio mimicus, Vibrio cholerae, Vibrio penaeicida and other bacterial species tested. MAbs in groups 1, 2 and 3 were able to use for the detection of bacterial infection in the tissues by means of immunohistochemistry.
MAbs specific to V. alginolyticus was produced. These MAbs can be used for specific identification of the bacteria by simple 'dot blotting' method and immunohistochemistry.
This study demonstrated an immunological tool that can be used for simple and accurate identification of V. alginolyticus as well as for the diagnosis of V. alginolyticus infection in animals. This immunological tool can replace costly and laborious biochemical tests.
本研究旨在制备单克隆抗体（MAbs），用于简单、特异性地鉴定虾类溶藻弧菌感染。
用热灭活的溶藻弧菌对小鼠进行免疫，每隔2周免疫一次，共免疫4次。选用免疫反应最佳的小鼠作为杂交瘤制备的脾细胞供体。通过对溶藻弧菌和其他细菌进行斑点印迹、蛋白质免疫印迹以及对感染虾组织进行免疫组织化学分析，筛选产生所需抗体的杂交瘤克隆。获得了四组单克隆抗体；第一组单克隆抗体仅对用于免疫的溶藻弧菌表现出有限的特异性，而第二组和第三组识别用于测试的所有三株溶藻弧菌。第四组单克隆抗体与所有三株溶藻弧菌结合，还识别副溶血性弧菌、哈维氏弧菌、河流弧菌和创伤弧菌，但不与模仿弧菌、霍乱弧菌、对虾弧菌及其他测试细菌结合。第1、2和3组的单克隆抗体能够通过免疫组织化学方法检测组织中的细菌感染。
制备了针对溶藻弧菌的单克隆抗体。这些单克隆抗体可通过简单的“斑点印迹”法和免疫组织化学用于该细菌的特异性鉴定。
本研究展示了一种免疫工具，可用于简单、准确地鉴定溶藻弧菌以及诊断动物的溶藻弧菌感染。这种免疫工具可替代昂贵且费力劳神的生化检测。
