Determination of T-2 Toxin and HT-2 Toxin in Milk: A Comparison of Three Formats of Immunoassays

Abstract

This research paper presents a comparative study of three formats of immunoassays, including conventional enzyme-linked immunosorbent assay (ELISA), one-step ELISA, and reverse enzyme-linked immunosorbent assay (reELISA), for the determination of T-2 toxin (T-2) and HT-2 toxin (HT-2). This comparative study was performed with regard to specificity, sensitivity, matrix effect, accuracy, and precision. Among the three procedures, reELISA exhibited the best sensitivity with limits of detection (LOD) at 3.04 and 2.79 ng mL⁻¹ for T-2 and HT-2. For the other parameters, there were no apparent differences. Good recoveries in negative spiked samples at concentrations of 50, 200, and 500 ng mL⁻¹ were obtained, and the values were 67.9–112.9% for conventional ELISA, 60.8–108.8% for one-step ELISA, and 62.7–88.1% for reELISA. The relative standard deviation (RSD) for each procedure was less than 10.3%, 14.0%, and 11.5%, respectively. Among all three methods, the one-step protocol was the most time efficient, and the reELISA procedure exhibited the best sensitivity.

Keywords:

• Enzyme-linked immunosorbent assay
• HT-2 toxin
• Method comparison
• T-2 toxin

Acknowledgments

Yanshen Li and Yanrong Zhang contributed equally.

This work was financially supported by the National Basic Research Program of China (973 Program; Grant No.2009CB118801), and the Chinese-Belgian Joint Project of BELSPO, Belgium (BL/02/C58) and MOST, China (S2012GR0016).

Notes

RSD values are calculated by average of five determination (n = 5) of three successive day.