大型溞ChE和NAGase间接竞争和非竞争ELISA方法的比较

[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/m L和1∶10 000,灵敏度为7.426 7 ng/m L;间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为5.277μg/m L和1∶8 000,灵敏度为0.163 4 ng/m L。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8.961μg/m L和1∶6 000,灵敏度为4.199 9 ng/m L;间接竞争ELISA的最适抗原浓度和抗体稀释倍数分别为6.450μg/m L和1∶4 000,灵敏度为0.250 8 ng/m L。[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。

Comparison of Competitive and Non-competitive ELISA for Determination of ChE and NAGase from Daphnia magna

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.