食品科学

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黄曲霉毒素B1酶联免疫试剂盒稳定性研究

韩春卉,江 涛*,李燕俊,高秀芬,李 楠,张宏元,赵 熙,张 靖   

  1. 国家食品安全风险评估中心,卫生部食品安全风险评估重点实验室,北京 100021
  • 出版日期:2013-07-25 发布日期:2013-08-02

Stability of Aflatoxin B1 ELISA Kit

Han chun-hui,Jiang tao*,Li yan-jun,Gao Xiu-fen,Li nan,Zhang hong-yuan,Zhao xi,Zhang jing   

  1. Key Laboratory of Food Safety Risk Assessment, Ministry of Health, China National Center for Food Safety Risk Assessment,Beijing 100021, China
  • Online:2013-07-25 Published:2013-08-02

摘要:

目的:研究自制黄曲霉毒素B1(AFB1)酶联免疫试剂盒在不同温度下的稳定性。方法:比较研制出的AFB1酶联免疫试剂盒在4℃和37℃保存条件下加入不同质量浓度的AFB1后其空白对照孔OD值的变化范围、IC20、IC50、IC80及RSD值的变化情况,及标准曲线的R²值等指标。结果:4℃保存270d空白对照孔OD值变化范围为1.129~1.777,IC20、IC50、IC80及RSD分别为0.05ng/mL和27.36%、0.27ng/mL和27.58%、0.92ng/mL和19.82%,该保存时间内各标准曲线线性关系良好(R²值均大于0.97);37℃加速破坏条件下保存13d内空白对照孔OD值变化范围为1.200~1.809,IC20、IC50、IC80及RSD变化分别为0.06ng/mL和27.71%、0.25ng/mL和25.66%、1.181ng/mL和21.89%,该保存时间内各标准曲线线性关系良好(R²值均大于0.98)。结论:研制的AFB1酶联免疫试剂盒在4℃和37℃加速破坏条件下分别保存270d和13d,试剂盒空白对照孔的OD值均大于1.0,各项指标可满足现场检测要求。

关键词: 黄曲霉毒素B1, 酶联免疫试剂盒, 稳定性

Abstract:

Objective: To explore the effect of storage temperature on the stability of the laboratory-fabricated enzyme linked
immunosorbent assay (ELISA) kit for AFB1 detection. Methods: Changes in OD value of blank control wells, IC20, IC50 and
IC80 (the concentrations required for 20%, 50% and 80% inhibition, respectively) and RSD were examined upon addition of
different concentrations of AFB1 when stored at 4 or 37 ℃ and the R² values of the standard curves at different storage times
were investigated. Results: On the 270th day of storage at 4 ℃, the OD value of blank control wells varied from 1.129 to 1.777
and the IC20, IC50 and IC80 were 0.05, 0.27 ng/mL and 0.92 ng/mL, with RSD of 27.36%, 27.58% and 19.82%, respectively. The
standard curves developed at different time points during the 270 d storage period exhibited good linearity (R² greater than 0.97).
The OD value of blank control wells varied from 1.200 to 1.809 over a storage period of 13 d under the accelerated conditions
at 37 ℃ and the IC20, IC50 and IC80 were 0.06, 0.25 ng/mL and 1.181 ng/mL with RSD of 27.71%, 25.66% and 21.89%,
respectively. Similarly, the standard curves developed at different time points during the 270 d storage period also showed good
linearity (R² greater than 0.98). Conclusion: The OD value of blank control wells in the ELISA kit remains above 1.0 regardless
of storage at 4 ℃ for 270 d or 37 ℃ for 13 d and all indexes examined can meet the requirements for in situ detection.

Key words: aflatoxin B1 (AFB1), enzyme linked immunosorbent assay (ELISA) kit, stability