1 Ton DucThang University, Vietnam
2 Sac Ky Hai Dang science technology services joint stock conpany, Vietnam
3 University of Natural Science, Vietnam
Citation: Bui QA, Tong D, Chu PN, Hoang CL (2019) Develop the first elisa kit detecting melamine residues in feed and milk products in Vietnam. SciTech Immuno-Microbiology 2019. Dubai: UAE
Received: April 02, 2019 Accepted: April 04, 2019 Published: April 04, 2019
The adulteration of melamine into food is banned but its misuse has been widely reported in milk products and animal feed. The development of a rapid screening immunoassay for monitoring this substance is an urgent requirement. In this research, a hapten of melamine was successfully synthesized by introducing six carbon chain length spacer arm via a reaction between 6-aminocaproic acid and 2-chloro-4,6-diamino-1,3,5-triazine(CAAT). The molecular structure of this hapten was indentified by H1 nuclear magnetic resornance spectrometry. An immunogen was prepared by coupling this hapten to Keyhole limpet hemocyanin (KLH). An enzyme conjugated was prepared by coupling synthesizedhapten to Horse raddish peroxidase (HRP). A direct competive ELISA (cELISA) was developed to evaluate assay performance. The results showed that high titer polyclonal antibodies were obtained, and the performance of this cELISAkit showed good specification: IC50 of 230 ng mL-1, a LOD of 8.3 ng mL-1, and LOQ of 10ng mL-1. Regarding selectivity performance, no obvious cross-reactivity to common compound was found. This data showed that this ELISA kit met the requirement of melamine maximum residues level (1000 ng mL-1 in milk products, 2500 ng mL in animal feed), and could be use in an immunoassay for the rapid and sensitive detection of this banned chemical.