Chemiluminescent Characteristic of a New Type of Enhanced Chemiluminescent Substrate Solution and It’s Application in Immunoassay
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摘要: 在辣根过氧化物酶 /鲁米诺 /增强剂 /过氧化氢 (HRP/luminol/enhancer/H2 O2 )增强化学发光体系中, 加入一系列辅助试剂, 获得了一种灵敏度高、发光持续稳定、可长期贮存的新型酶促化学发光增强液。该增强液灵敏度为 2 4amol/孔, 发光半衰期为 4 0min, 工作液在室温可稳定存放 1个月, 在 37℃下可稳定存放 14d。在此基础上, 在TSH BASECLEIA和AFPECLEIA上进行了应用。TSH BASE CLEIA测量范围为 0 17～ 35mIU/L, 信噪比大于 5;分析灵敏度为 0 0 4mIU/L, 功能灵敏度为0 0 6mIU/L 批内变异系数为 2 96 %～ 5 0 5%, 批间变异系数为 4 0 1%～ 7 56 % 回收率为 10 1%～119% 稀释实验相关方程为y =4 4.6 50x + 0. 352, r =0. 996 与BayerACS :180系统进行方法学比较, 相关方程为 y=0. 889x - 0. 351,r =0. 984。AFPECLEIA测量范围为 5～ 2 0 0 0ng/mL, 信噪比大于 4。动力学实验表明, 加入发光液之后 15min测量, 结果准确可靠。
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关键词:
- 辣根过氧化物酶(HRP) ,
- 鲁米诺(luminol) ,
- 增强剂(enhancer) ,
- 增强化学发光酶免疫分析(ECLEIA) ,
- 促甲状腺素(TSH)
Abstract: A series of assistant reagents were added to the horseradish peroxidase (HRP)/luminol/enhancer/H 2O 2 system, and got a new type of enhanced chemilumine- scent substrate solution catalyzed by HRP. It has the advantage of high sensitivity, slow decay of light emission, being stored for long time, and so on. The assay sensitivity for HRP is 2.4 amol per well. The half-life of decay of light emission is about 40 min. The work solution can be stored for more than 14 d at 37 ℃ or 30 d at room temperature. A two-site sandwich BAS ECLEIA for TSH and a two-site sandwich ECLEIA for AFP were developed. In the TSH-BAS ECLEIA, the range of detection is 0.17～35 mIU/L. The signal to background ratio is above 5.The assay sensitivity and function sensitivity are 0.04 mIU/L and 0.06 mIU/L, respectively. The intra-assay and inter-assay coefficients of variation are 2.96%～5.05% and 4.01%～7.56%, respectively. The recovery is 101%～119%. The correlation coefficient and the correlation equations of sample dilution are 0.996, y=44.65x+0.352. The correlation coefficient and the correlation equations of assay with Bayer ACS:180 system are 0.984, y= 0.889x-0.351. In the AFP ECLEIA, the range of detection is 5～2 000 ng/mL. The signal to background ratio is above 4. The kinetics experiment shows that the result is accurate and reliable in 15 min after initiation the chemiluminescent reaction. -
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