摘要
[目的]探索间接非竞争ELISA和间接竞争ELISA法测定大型溞ChE和NAGase的试验条件及方法灵敏度。[方法]分别采用间接竞争和间接非竞争ELISA方法测定大型溞ChE和NAGase。[结果]对于ChE，间接非竞争ELISA的最适抗原浓度和抗体稀释倍数分别为1．466μg／mL和1∶10000，灵敏度为7．4267 ng／mL；间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为5．277μg／mL和1∶8000，灵敏度为0．1634 ng／mL。对于NAGase，间接非竞争ELISA 的最适抗原浓度和抗体稀释倍数分别为89．61μgm／L 和1∶6000，灵敏度为41．999 ng／mL；间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为6．450μg／mL和1∶4000，灵敏度为0．2508 ng／mL。[结论]无论是ChE还是NAGase，间接竞争ELISA在最适抗体浓度下的检测灵敏度均高于间接非竞争ELISA，加之其操作简便，间接竞争ELISA在生物和环境样品ChE和NAGase含量检测中值得推广应用。
Abstract
Objective] To explore the test condition and method sensitivity of Cholinestersae (ChE) and β-N-Acetyl-D-glucosaminidase (NA-Gase) from Daphnia magna by the methods of competitive and non-competitive ELISA.[Method] ChE and NAGase of Daphn ia magna were de-tected by the methods of indirect competitive and non-competitive ELISA,respectively.[ Result] As for ChE,the optimal dilutions for the antigen and the antibody were 1.466 μg/mL and 1∶10 000,respectively, and the sensitivity was measured to be 7.426 7 ng/mL if the enzyme was tested by indirect non-competitive ELISA.In case that the enzyme was tested by indirect and competitive ELISA,the optimal dilutions for the antigen and the antibody were 5.277 ug/mL and 1∶8 000,respectively,and the sensitivity was 0.163 4 ng/mL.As for the NAGase,the optimal dilutions for the antigen and the antibody were 8.961μg/mL and 1∶6 000,respectively,and the sensitivity was 4.199 9 ng/mL if the enzyme was tested by the indirect and non-competitive ELISA.In case that the enzyme was test by indirect and competitive ELISA,the optimal dilutions for the antigen and the antibody were 6.450 μg/mL and 1∶4 000,respectively,and the sensitivity was 0.250 8 ng/mL.[Conclusion] Indirect and competitive ELISA is more sensitive than the non-competitive ELISA under the optimal concentration antigen for both ChE and NAGase.Moreover,it is simple in op-eration.Thus,indirect and competitive ELISA is worth to be promoted in determining NAGase and ChE content in biological and environmental samples.
国家科技期刊平台
NSTL
维普
万方数据