| 摘要: |
| 【目的】建立黄颡鱼仔鱼(Pelteboagrus fulvidraco)卵黄蛋白的ELISA检测方法，探讨黄颡鱼仔鱼体内卵黄蛋白含量的发生规律及仔鱼投喂生物饵料的最佳时间，为研究黄颡鱼仔鱼早期营养和开口饵料提供依据【方法】以黄颡鱼仔鱼为试验材料，卵黄蛋白抗血清为抗体，纯化的卵黄蛋白为抗原，建立间接竞争酶联免疫吸附反应(ELISA)检测方法，测定黄颡鱼仔鱼卵黄蛋白在不同发育时期（破膜43，48,57,65,72,81,89,96,105,113,120和129 h）的变化情况。【结果】黄颡鱼仔鱼卵黄蛋白ELISA检测中，抗血清的最佳稀释倍数为1∶20 000倍,包被抗原的最佳质量浓度为125 ng/mL，最适检测质量浓度为20～2 000 ng/mL,批内变异系数为6.529%，批间变异系数为6.873%。利用建立的ELISA方法检测发现，黄颡鱼卵黄蛋白含量随发育时间的延长而逐渐递减，至96 h时缓慢下降直至消失。【结论】利用ELISA方法能相对准确地检测黄颡鱼仔鱼卵黄蛋白含量的变化，破膜96 h是黄颡鱼仔鱼由内源营养阶段转向外源营养阶段的转折点，此时开始投喂适口的生物饵料更有利于黄颡鱼仔鱼开口摄食及生长发育。 |
| 关键词: 黄颡鱼仔鱼 卵黄蛋白 ELISA检测 |
| DOI： |
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| 基金项目:国家自然科学基金项目（30671621）;吉林省科技发展计划项目（吉科合字20040208-1） |
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| ELISA detection method for vitellin of yellow catfish(Pelteboagrus fulvidraco)larvae |
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HUANG Quan, WANG Weijie, ZHANG Dongming
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Jilin Agricultural University
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| Abstract: |
| 【Objective】This study intended to establish a vitellin ELISA method for exploring the occurrence of vitellin content and the best time of larvae feeding live prey for yellow catfish.【Method】Yellow catfish larvae (Pelteboagrus fulvidraco)was test material.Antisera was used as antibodies and purified vitellin of yellow catfish was used as antigen to establish indirect competitive enzyme-linked immunosorbent assay the reactions (ELISA) to determine the changes in vitellin of yellow catfish in different developmental stages (43，48,57,65,72,81,89,96,105,113,120 and 129 h after break of membranes).【Result】The optimal working concentrations of antiserum and coating antigen were 1∶20 000 dilution and 125 ng/mL of vitellin,optimum detection ranged 20－2 000 ng/mL.Within-assay coefficient of variation was 6.529% and inter-assay coefficient of variation was 6.873%.The established ELISA detection method found that the vitellin content gradually decreased as the extension of the development time and disappeared after 96 h.【Conclusion】ELISA method can accurately detect the changes in vitellin content of yellow catfish.96 h after rupture of membranes was the turning point of the yellow catfish larvae from endogenous vegetative stage to exogenous nutrition stage and it was the best time to feed with live prey and improve the eating and growth. |
| Key words: yellow catfish (\%Pelteboagrus fulvidraco\%) larvae vitellin ELISA detection |
