| 溶藻弧菌单克隆抗体的制备及应用 |
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| 引用本文： | 宋晓玲.溶藻弧菌单克隆抗体的制备及应用[J].水产学报,2001,25(6):522-527. |
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| 作者姓名： | 宋晓玲 |
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| 作者单位： | 中国水产科学研究院黄海水产研究所,山东青岛,266071 |
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| 基金项目： | 中国水产科学研究院科研基金资助(95-011号) |
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| 摘 要： | 小鼠骨髓瘤细胞系SP2/0与经溶藻弧菌(1.1587)免疫的BALB3/C雌鼠脾细胞在PEG条件下融合,有45.8%的培养孔有杂交瘤细胞生长,其培养上清液抗体阳性率为77.4%.经反复有限稀释法克隆杂交瘤细胞,获得7株抗溶藻弧菌的单克隆抗体杂交瘤细胞株,分别为AC1-C9,AC1-C11,BB4-D4,AD1-A3,AD1-F3,AD2-E7,AD2-F7.取AD1-F3扩大培养,注射小鼠,制备了抗溶藻弧菌的单克隆抗体腹水,滴度为11000.该腹水与其他3株溶藻弧菌菌株有强交叉反应,与其他13株标准菌株无交叉反应.利用制备的单抗腹水,建立了检测溶藻弧菌的单抗-ELISA技术.该反应系统可应用于溶藻弧菌的快速诊断,反应时间为6-7h,检测灵敏度为104cells·-1.并利用该检测方法进行了11份待测菌样本溶藻弧菌的检测.
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| 关 键 词： | 单克隆抗体 溶藻弧菌 酶联免疫吸附试验 对虾病害 |
| 文章编号： | 1000-0615(2001)06-0522-06 |
| 收稿时间： | 2014/4/14 0:00:00 |
| 修稿时间： | 2001年3月2日 |
Production and application of monoclonal antibodies of Vibrio alginolyticus |
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| SONG Xiao ling.Production and application of monoclonal antibodies of Vibrio alginolyticus[J].Journal of Fisheries of China,2001,25(6):522-527. |
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| Authors: | SONG Xiao ling |
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| Institution: | (Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Science, Qingdao 266071,China |
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| Abstract: | Monoclonal antibodys (Mab) raised against Vibrio alginolyticus strains (1. 1587) by that primed spleen
cellsfusedwithSP2/0myelomacellsusingpolyethylingglycol. Therateoffusionwas 45.8%andtherateof
positiveinfusionwas 77.4%byELISA. Mabs, AC1-C9,AC1-C11 ,BB4-D4,AD1-A3,AD1-F3,AD2E7 ,AD2 -F7 , were harvested using usable hybridomas clone twice by limiting dilution. Ascites with Mabs against
Vibrio alginolyticus were harvested by injecting BALB/ C mice. The titer of the ascites was 1 : 1000. In
comparison , Mab AD1 -F3 reacted with only four Vibrio alginolyticus strains . No cross2reactions were observed
among thirteen non2Vibrio alginolyticus. Using this Mab ascites , an indirect Enzyme2Linked Immunosorbent
Assay (ELISA) for rapid diagnosis of Vibrio alginolyticus has been developed. This method can detect 1 ×104
bacteria per ml with in 6 -7 h. After the method was optimized , 11 samples were detected for Vibrio
alginolyticus. |
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| Keywords: | monoclonal antibodies(Mab) Vibrio alginolyticus Enzyme Linked Immunosorbent Assay(ELISA) |
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