Hao Gui-jie, Shen Jin-yu, Cao Zheng, Pan Xiao-yi
Institute of Freshwater Fisheries of Zhejiang, Huzhou 313001, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Sep;23(9):838-40.
To prepare the mAb against Vibrio harveyi and analyse their biological properties.
8-week-old female BALB/c mice were immunized with pathogenic V.harveyi GYC1108-1, which was isolated from the diseased great yellow croaker in Zhejiang China in 2003. Spleen cells collected from the immunized mice after the fifth immunization were fused with Sp2/0-Ag-14 myeloma cells. ELISA was used to screen hybridoma cells and identify the specificity and sensitivity of mAbs.
Limited dilution method was used to subclone the positive clones. After three cycles of subcloning, 5 mAbs against GYC1108-1 were obtained and were proved to have high titer of ELISA. The test of specificity suggested that 1B7 reacted on V.harveyi, V.alginolyticus and V.parahaemolyticus but did not react on V.mimicus, V.anguillarum, Aeromonas hydrophila and the other isolates among the 14 bacterium species tested. 1B7 was used to detect V.harveyi at the concentration of 10(4) CFU/mL by indirect ELISA.
The obtained mAbs could be used to diagnosis and prevent of V.harveyi in aquaculture rapidly.
