Development of an Ultra-sensitive Chemiluminescence Enzyme Immunoassay for the Determination of Diethylstilbestrol in Seafood

Abstract

An ultra-sensitive indirect competitive chemiluminescence enzyme immunoassay was developed for screening diethylstilbestrol in fish and shrimp samples. The concentration of diethylstilbestrol that caused 50% inhibition of the binding enzyme marker (IC₅₀) was 0.32 ng/mL and the limit of detection was 0.0068 ng/mL; the linear range was from 0.028 ng/mL to 3.60 ng/mL. The assay showed cross-reactivity of 7.1% and 2.8% with dienestrol and hexoestrol, respectively, but negligible cross-reactivity with estradiol, estrone, ethinyloestradiol, and progestin. The recovery from spiked fish and shrimp samples varied from 68.5% to 92.5%, and the mean coefficients of variation within groups and between groups were 6.2% and 8.0%, respectively. Our results indicated that the assay is a simple, sensitive, specific, and accurate method for screening fish and shrimp samples for diethylstilbestrol.

Keywords:

• Chemiluminescence enzyme immunoassay (CLEIA)
• Diethylstilbestrol
• Seafood

Acknowledgments

Jin Yi Yang and Yan Zhang represent equal contributors.

This work was supported by National Natural Science Foundation of China (31201361), the Science and Technology Project of Guangzhou City (11BppZXcc2100021), the Science and Technology Project of Guangdong Province (2010A032000001-4, 2012A020100002), the National Spark Program (2012GA780001), and the Science and Technology Project of Guangzhou City (Zhu Jiang Xin Xing) (1217000380).

Notes

LOD: Limit of detection.

^a LOD is 0.0068 ng/mL.

^b LOD is 0.041 ng/mL.