王莹 白晶晶 李玉芳 钱鹏 曹玉姣 陈凌燕 郝丽影 邓均华 田克恭

摘要 制备猫泛白细胞减少症病毒（FPV）单克隆抗体，研制胶体金检测试纸条。用FPV临床分离株免疫Balb/c小鼠，取血凝抑制（HI）效价最高的小鼠脾细胞与SP2/0细胞融合，采用HI方法筛选获得2株可稳定分泌FPV单克隆抗体的杂交瘤细胞，并对其用于胶体金检测试纸条进行评价。2株杂交瘤细胞3C3和4A1株，分泌的单克隆抗体重链亚型分别为IgG2b、IgG2a，轻链亚型均为kappa;腹水纯化后单克隆抗体3C3、4A1的HI效价分别为1∶5 120、1∶10 240。用胶体金标记单克隆抗体4A1，单克隆抗体3C3作为检测线，羊抗鼠二抗作为对照线制备的胶体金检测试纸条，检测FPV病毒液的灵敏度为103.7TCID50/mL;检测猫源其他病毒如FHV-1、FCV均为阴性;批内和批间重复性检测不同含量的FPV结果一致;检测513份临床样品，与PCR的阳性符合率为76%（71/93），阴性符合率为100%（420/420），总符合率为96%（491/513）。该研究制备的FPV单克隆抗体可用于FPV胶体金检测试纸条的生产，用于FPV的现场快速检测。

关键词 猫泛白细胞减少症病毒;单克隆抗体;胶体金检测试纸条

中图分类号 S 855.3  文献标识码 A  文章编号 0517-6611（2021）15-0099-05

Abstract Feline panleukopenia virus （FPV） monoclonal antibodies （MAbs） were developed for the preparation of colloidal gold test strips. Balb/c mice were firstly immunized with FPV clinical strain， and then the spleen cells of the mouse with the highest hemagglutination inhibition （HI） titers were fused with myeloma cells SP2/0. Hybridoma cells 3C3 and 4A1 strains were screened by HI and chosen for the preparation of immunochromatographic test strips. The HI titers of the MAbs from purified ascites 3C3 and 4A1 are 1∶5 120 and 1∶10 240， and the heavy chain subtypes were IgG2b and IgG2a selectively， the light chain subtypes were both kappa. The MAb 4A1 was conjugated with colloid gold， the 3C3 was coated on a porous nitrocellulose membrane as the detection line， and the Goat antirat IgG antibody was coated as the control line， respectively. The detection limit of the strip was 103.7TCID50/mL and no crossreaction occurred between FPV and other feline viruses. Intraassay and interassay repeatability test results were consistent. 513 clinical samples were tested by both the test and PCR. The result showed that the positive coincidence rate of the test was 76% （71/93） with PCR， the negative coincidence rate was 100% （420/420）， and the overall consistency rate was 96% （491/513）. The MAbs 3C3 and 4A1 prepared in this study could be used for the preparation of colloidal gold test strip.

Key words Feline panleukopenia virus;Monoclonal antibody;Colloidal gold immnuochromatographic assay

基金項目 洛阳市河洛英才计划“新型兽用生物制品研发及产业化”。

作者简介 王莹（1986—），女，河南新乡人，高级工程师，硕士，从事诊断试剂研究;白晶晶（1987—），女，河南巩义人，助理工程师，从事诊断试剂研究。王莹和白晶晶为共同第一作者。

*通信作者：邓均华，兽医师，硕士，从事动物疫苗和诊断试剂研究;田克恭，研究员，博士，从事动物疫病诊断和防控技术研究。

收稿日期 2021-01-01

猫泛白细……