Wang Cai-Xia, Wang Guo-Ping, Hong Ni, Jiang Bo, Liu Hui, Wu Kang-Wei
National Indoor Conservation Center of Virus-free Gemplasms of Fruit Crops, Wuhan 430070, China.
Sheng Wu Gong Cheng Xue Bao. 2006 Jul;22(4):629-34.
Citrus tristeza virus (CTV) was purified from a citrus sample by a modified protocol, and the yield was about 1 mg from 100 g citrus tissues. Polyclonal antibody was prepared by immunizing rabbits with the purified CTV preparation with a titer 1:25600 in indirect ELISA test. Eighteen hybridoma-cell lines secreting monoclonal antibodies (MAbs) against CTV were screened after the fusion of mouse myeloma cells (SP2/0) with spleen cells from BALB/c immunized with the virus preparation. Four hybridoma-cell lines were selected randomly for later analysis. The results indicated that the titers of ascetic fluids against these hybridoma cell lines ranged from 1:51200 to 1:204800 in indirect ELISA, and their isotypes and subclasses were IgG2a for 2G and 3H and IgG2b for IE and 4H. These four Mabs were used to detect CTV in citrus samples in different sources. Results showed that TAS-ELISA with polyclonal antibody as trapping antibody and monoclonal antibody as testing antibody had a higher specificity and sensitivity than PAS-ELISA. Four Mabs showed different intensities of serological reaction with different CTV isolates. However, much work remains for realizing the characteristics and the serological relationships among these isolates.
采用改良方案从柑橘样品中纯化柑橘衰退病毒（CTV），100克柑橘组织的产量约为1毫克。用纯化的CTV制剂免疫兔子制备多克隆抗体，其在间接ELISA试验中的效价为1:25600。将小鼠骨髓瘤细胞（SP2/0）与用病毒制剂免疫的BALB/c小鼠的脾细胞融合后，筛选出18株分泌抗CTV单克隆抗体（MAb）的杂交瘤细胞系。随机选择4株杂交瘤细胞系用于后续分析。结果表明，间接ELISA法检测这些杂交瘤细胞系腹水的效价范围为1:51200至1:204800，其亚型和亚类中，2G和3H为IgG2a，IE和4H为IgG2b。用这4种单克隆抗体检测不同来源柑橘样品中的CTV。结果表明，以多克隆抗体为捕获抗体、单克隆抗体为检测抗体的TAS-ELISA比PAS-ELISA具有更高的特异性和敏感性。4种单克隆抗体与不同CTV分离株的血清学反应强度不同。然而，要了解这些分离株的特性和血清学关系，仍有许多工作要做。
