家蚕微孢子虫单抗金银染色法检测技术的研究

使用新构建的抗家蚕微孢子虫（N．b）的单克隆抗体，结合免疫金银染色（IGSS），进行检测N．b。设计6种温度和5种时问，对N．b等9种微孢子虫检测，结果表明，该细胞株单抗直接IGSS法最佳工作条件：胶体金标记pH值6.2～6.5，金标抗体染色37℃、保湿30min，经显影后光镜观察，N．b孢子周边被染上棕褐色，其它类型孢子呈无色或浅褐色；实验还比较了抗原涂片后，不同物理、化学方法固定，结果甲醇固定效果较好。

STUDIES ON THE MONOCLONAL ANTIBODY BASED IMMUNOGOLD-SILVER STAINING METHOD FOR DETECTING SPORES OF Nosema bombycis

The study was employed to detect Nosema bombycis(N. b)by the method of monoclonal antibodies (McAb)based immunoglod-silver staining (IGSS). The McAb was prepared and purified from the newly developed cell lines to N. b spores. Six temperatures and five time degrees were tested and the results of detecting nine kinds of microsporidians showed that the optimum staining conditions of direct McAb-IGSS assay was 37℃,30 min and PH 6.2~ 6.5. By the staining,N. b spores were dyed dark brown,the others were not dyed or dyed light brown. Moreover,the influence of different fixing methods on the morphology of the antigens was examined,and it showed that good staining results can be obtained by methanol fixative.