牛奶中替米考星间接竞争酶联免疫吸附检测法的建立

doi:10.16431/j.cnki.1671-7236.2015.10.017

Abstract

Abstract: An indirect competitive enzyme-linked immunosorbent assay (ELISA) for detecting tilmicosin residue in milk was developed and applied to milk samples after the determination of dilution of coating antigen and antibody to timicosin by using checkerboard method,and the optimization of coating condition,reaction time and temperature,and reaction time of enzyme-labled secondary antibody.The results showed that the 50% inhibition concentration (IC₅₀) of the indirect competitive ELISA was 2.1 ng/mL and the limit of detection (LOD) of tilmicosin in milk was 2 μg/L.The recoveries of tilmicosin added in milk at 5,10 and 20 μg/L ranged from 79.2% to 90.1% with the coefficients of variation (CV) not higher than 9.0%.The indirect competitive ELISA would not react with other common macrolide drugs.The sensitivity of indirect competitive ELISA developed in the study complied to the maximum residue limit of tilmicosin set by China,and laid the foundation for the test kit of the rapid detection of tilmicosin in milk.

Key words: indirect competitive enzyme-linked immunosorbent assay (ELISA); tilmicosin; milk; residue

CLC Number:

S852.4

HAN Qing, WANG Jing, XU Hao, YANG Juan. Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay for Tilmicosin in Milk[J]. , 2015, 42(10): 2625-2630.

References

[1] 江海洋,沈建忠,胡顶飞.兽用抗菌药物替米考星[J].中国兽药杂志,2002,36(8):28-31.
[2] 郑亚秋,曹湛,郭宏斌,等.替米考星分子印迹聚合物的制备及其固相萃取研究[J].分析化学,2010,38(1):95-99.
[3] 李存,沈建忠,江海洋,等.猪组织中替米考星残留的高效液相色谱检测方法研究[J].畜牧兽医学报,2005,36(10):1075-1078.
[4] 王海涛,张睿,段宏安,等.高效液相色谱法同步检测牛奶中替米考星、泰乐菌素和螺旋霉素残留量[J].分析试验室,2008,27(7):98-101.
[5] Cooper J,Delahaut P,Fodey T L,et al.Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA[J].Analyst,2004,129:169-174.
[6] Jeon M,Kim J,Paeng K J,et al.Biotin-avidin mediated competitive enzyme-linked immunosorbent assay to detect residues of tetracyclines in milk[J].Microchem J,2008,88:26-31.
[7] Cliquet P,Cox E,Haasnoot W,et al.Extraction procedure for sulfachloropyridazine in porcine tissues and detection in a sulfonamide-specific enzyme-linked immunosorbent assay (ELISA)[J].Anal Chim Acta,2003,494:21-28.
[8] 罗晓琴,万宇平,孙震,等.牛奶中泰乐菌素和替米考星的胶体金免疫层析法测定[J].中国乳品工业,2012,40(10):42-45.
[9] 朱事康,聂继斌,刘星,等.动物源性食品中泰乐菌素残留ELISA试剂盒的研制[J].中国兽药杂志,2009,8:5-8.
[10] Zhang J K,Liu J X,Wang L L,et al.Production of the monoclonal antibody against tylosin for immunoassay of macrolide antibiotics in milk[J].J Environ Sci Health,2012,47(9):876-882.
[11] Burkin M,Galvidis I.Simultaneous separate and group determination of tylosin and tilmicosin in foodstuffs using single antibody-based immunoassay[J].Food Chem,2012,132(2):1080-1086.
[12] Le T,He H Q,Niu X D,et al.Development of an immunochromatographic assay for detection of tylosin and tilmicosin in muscle,liver,fish and eggs[J].Food Agric Immunol,2013,24(4):467-480.
[13] Wang Z H,Mi T J,Zhang H Y,et al.Hapten synthesis,monoclonal antibody production,and development of a competitive indirect enzyme-linked immunosorbnent assay for erythromycin in milk[J].Food Chem,2015,171:98-107.
[14] Dickson L C,O'Byrne C,Chan W.A quantitative method for residues of macrolide antibiotics in porcine kidney by liquid chromatography/tandem mass spectrometry[J].J AOAC Int,2012,95:567-575.
[15] Jiang W,Wang Z,Beier R C,et al.Simultaneous determination of 13 fluoroquinolone and 22 sulfonamide residues in milk by a dual-colorimetric enzyme-linked immunosorbent assay[J].Anal Chem,2013,85:1995-1999.
[16] Zhang Y,Lu S,Liu W,et al,Preparation of anti-tetracycline antibodies and development of an indirect heterologous competitive enzyme-linked immunosorbent assay to detect residues of tetracycline in milk[J]. J Agric Food Chem,2007,55:211-218.

Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay for Tilmicosin in Milk

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	MA Xiaojiao, ZHAO Yankun, SHAO Wei, WU Yating, LI Ming, LIU Huimin, MENG Lu, CHEN He. Isolation,Identification and Drug Resistance Analysis of Staphylococcus aureus in Raw Milk in Some Areas of Xinjiang [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 789-797.
[2]	WU Tao, QI Yunxia, LIU Wei, ZHU Honglong, HUANG Dongwei, DING Haisheng, ZHAO Huiling, CHENG Guanglong, ZHAO Xiaowei. Effect of Colostrum and Milk Feeding on Hepatic Proteins of Calves Based on Proteomics Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 37-45.
[3]	ZENG Zhiqin, LOU Wenqi, XU Jingyi, MA Yehua, ZHANG Yongfu, REN Xiaoli, CHEN Shaokan, YAN Lei, GUO Gang, ZHANG Zhen, WANG Yachun. Analysis of Impact Factors and Phenotype Prediction for Transforming Progesterone Concentration in Chinese Holstein Cows During Lactation [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 186-197.
[4]	XU Haoqi, XU Jingyi, HU Lirong, ZHANG Fan, LUO Hanpeng, ZHANG Hailiang, SHI Rui, LI Xiang, LIU Lin, LIU Qiaoxiang, GUO Gang, WANG Yachun. Polymorphism of PIK3CB Gene and Its Association Analysis with Reproductive and Milk Production Traits in Chinese Holstein Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3438-3452.
[5]	JI Haonan, GAO Qing, CHANG Yao, ZHANG Hailiang, WU Hongjun, CHAI He, YU Ying, WANG Yachun. Analysis of Influencing Factors on Milking Speed Indicators in Holstein Cows [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3475-3483.
[6]	TANG Qingfeng, LI Ling, HUANG Jiaxiang, FANG Wenyuan, NONG Haoru, TANG Yan, LU Chengwei, WEI Panqiu, FENG Ling. Survey on Mycotoxin Contamination of Buffalo Milk in Different Seasons Under Different Feeding Modes in Guangxi [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3665-3675.
[7]	ZHANG Zhenyu, WANG Yong. Advances on the Detection of Aflatoxins in Milk and Dairy Products [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2820-2830.
[8]	SUN Xingya, LI Qiang, WANG Zhanhui, YU Wenbo, SHI Weimin, ZHANG Suxia. Comparative Study on Veterinary Drug Residue Limit Standards for Poultry Meat and Eggs in China, EU and USA [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2362-2375.
[9]	SUN Xiaoshan, LIN Dongmei, SHAO Yuxin, WANG Zheng, HUANG Jianguo. Study on the Changes of Crude Protein and Mineral Elements in Pigeon Milk at Different Ages [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1697-1706.
[10]	LIU Jinhuan, ZHANG Shanling, SONG Wei, LUO Wanhe, CHEN Wei. Preparation of Quercetin-tilmicosin Mixture Nanoparticles and Its Antibacterial Activity Against Small Colony Variant Strains of Staphylococcus aureus [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1470-1478.
[11]	XU Liyun, ZHENG Menghe, XU Zihao, NIU Gaowei, WANG Yongli, QIU Hui, WU Yuxuan, JIN Tong, SHAO Wei. Effects of Astragalus Polysaccharide Supplementation on Milk Yield,Immune and Antioxidant Performance of Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3819-3828.
[12]	LIANG Shasha, PANG Chunying, DENG Tingxian, LU Xingrong, DUAN Anqin, MA Xiaoya, FANG Yanyan, LIANG Xianwei. Polymorphism of MFSD2A Gene and Its Association with Milk Production Traits in Buffalo [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3856-3865.
[13]	CHANG Lingling, TANG Qingping, LIU Jiajia, ZHANG Rui, FU Shengyong, MU Chunyu, BU Zhu. Analysis on Dynamic Changes of Nutrient Levels and Physiological Active Substances in Pigeon Milk at Different Periods of Squabs [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3215-3222.
[14]	SHI Fangquan, WANG Hui, ZHANG Fan, XIONG Benhai. Influencing Factors and Preventive Measures of Milk Fever in Cows [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3254-3263.
[15]	NING Jingyang, MA Longgang, CHEN Ziwei, AN Tao, CHANG Yao, WANG Kai, WANG Lei, CHEN Shaokan, ZHANG Fan, GUO Gang, ZHANG Hailiang, WANG Yachun. Population Characteristic of Dry Period Length and Its Relationship with Production Performance in Chinese Holstein Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3368-3377.