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ARS Home » Plains Area » College Station, Texas » Southern Plains Agricultural Research Center » Food and Feed Safety Research » Research » Publications at this Location » Publication #319484
Title: Analysis of pirlimycin residues in beef muscle, milk and honey by a biotin-streptavidin-amplified enzyme-linked immunosorbent assay

Author
item JIANG, WENXIAO - Shenzhen University
item Beier, Ross
item LUO, PENGJIE - Center For Food Safety Risk Assessment
item ZHAI, PENG - Shenzhen University
item WU, NAN - Center For Food Safety Risk Assessment
item LIN, GUIMIAO - Shenzhen University
item WANG, XIAOMEI - Shenzhen University
item XU, GAIXIA - Shenzhen University

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/15/2015
Publication Date: 12/15/2015
Publication URL: https://handle.nal.usda.gov/10113/61964
Citation: Jiang, W., Beier, R.C., Luo, P., Zhai, P., Wu, N., Lin, G., Wang, X., Xu, G. 2015. Analysis of pirlimycin residues in beef muscle, milk and honey by a biotin-streptavidin-amplified enzyme-linked immunosorbent assay. Journal of Agricultural and Food Chemistry. 64(1):364-370.

Interpretive Summary: Antimicrobial residues in the food supply can lead to allergic or toxic reactions and to an increase in antimicrobial resistance in pathogenic bacteria. Pirlimycin (PIR) is one of the most commonly used lincosamide antibiotics in human and veterinary medicine. PIR is used to treat Staphylococcus aureus infections and for the treatment of mastitis in lactating dairy cows. The incorrect use of PIR may leave residues in edible animal tissues and in milk. The immunoassay is advantageous for screening large numbers of samples. Immunoassays make use of antibodies and are used as an analytical tool to analyze for a specific compound. Antibodies are substances that are produced by the immune system in response to foreign substances which enter the body. Once the antibodies to a foreign substance are isolated, they can be used in a method to detect the presence of that foreign substance. Here we have improved the sensitivity of the standard immunoassay by reacting the antibodies with biotin, then by using a streptavidin labeled enzyme, a number of enzyme molecules will bind the antibody and cause an enhanced detection signal. This method will improve the sensitivity of the immunoassay over three times. This method is useful for measuring PIR residues in beef, milk, and honey, and can specifically be used for routine screening analysis of PIR residues in milk.

Technical Abstract: Food contamination caused by veterinary drug residues is a world-wide public health concern and requires continuous monitoring. In this paper, we describe a biotin–streptavidin-amplified ELISA (BA-ELISA) for detecting pirlimycin residues in beef, milk, and honey. The IC50 value of the BA-ELISA was 1.6 ng/mL for pirlimycin in buffer, and the sensitivity was improved 3 times when compared to traditional ELISAs. The optimized ELISA can be used to quantify trace amounts of pirlimycin residues in beef, milk, and honey. This method was sensitive with a limit of detection (LOD) of 4.45 µg/kg in beef, 1.65 µg/L in milk, and 2.75 µg/L in honey. The average recoveries of the BA-ELISA ranged from 77% to 97%, and the coefficient of variation ranged from 5.3% to 13.5%. The developed BA-ELISA method was validated using LC–MS/MS, and the BA-ELISA method can be used for routine screening analysis of pirlimycin residues in milk.