Preparation of Anti-Tetracycline Antibodies and Development of an Indirect Heterologous Competitive Enzyme-Linked Immunosorbent Assay to Detect Residues of Tetracycline in Milk

Zhang, Yulan; Lu, Shengxin; Liu, Wei; Zhao, Chengbiao; Xi, Rimo

doi:10.1021/jf062627s

Cited by 103 publications

(67 citation statements)

References 25 publications

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“…Therefore, rapid and simple detection techniques are required to determine the CPFX residue level in food products. Enzymelinked immunosorbent assay (ELISA) is very common as a biochemical and clinical analytical method, and is available for the detection of pesticides and veterinary drugs (Watanabe et al, 2001;Zhang et al, 2007;Peng et al, 2008;Stanker et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

Huang

Yin

Lü

et al. 2010

J. Zhejiang Univ. Sci. B

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Abstract:A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC 50 ) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%-84.60%, with coefficients of variation of less than 12.2%.

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Section: Introductionmentioning

confidence: 99%

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

Huang

Yin

Lü

et al. 2010

J. Zhejiang Univ. Sci. B

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“…The enzyme-linked immunosorbent assay (ELISA) technique is sensitive, specific and cost effective, and has been used diagnostically in recent years. Furthermore, ELISA methods have also been used for detection of various drug residues in the real system, such as in food and biological samples [12,13].…”

mentioning

confidence: 99%

“…The titer of the antibody was tested by an indirect competitive ELISA method [13]. The procedure was carried out as described below.…”

mentioning

confidence: 99%

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Meng

He³

et al. 2011

Chin. Sci. Bull.

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Based on the preparation of an anti-diethylstilbestrol (DES) monoclonal antibody, a simple and convenient indirect competitive enzyme-linked immunosorbent assay (ELISA) method for DES detection has been developed. The monoclonal antibody demonstrated high sensitivity to DES with an IC 50 value of 275 pg mL -1 and detection limit (LOD) of 90 pg mL -1 . The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol (<7%), estrone (<0.1%), estriol (<0.1%), and diethylstilbestrol benzoate (<0.1%). Chicken, fish, shrimp, urine and bile spiked with different concentration of DES were detected by the developed method, and the recovery rates were greater than 79.5%. Intra-and inter-assay variations were about 6%. This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples. The LODs in fish/shrimp, liver, feed and urine spiked with DES were 600, 600, 4800 and 600 pg mL -1 , respectively. These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals. Diethylstilbestrol (DES; Figure 1) is an orally active synthetic non-steroid estrogen. Since the 1960s, DES has been used as a growth-promoting agent in livestock [1,2]. Later, DES was found to cause cancer and its use was "phased out in the 1970s" by the FAO/WHO. In 1971, DES was reported to be a teratogen when given to pregnant women and has been associated with a rare vaginal cancer in female offspring [3]. On February 5, 1975, the US Food and Drug Administration (FDA) ordered 25 and 100 mg tablets of DES withdrawn; however, it was still illegally used in the livestock industry.*Corresponding authors (email: xirimo2000@yahoo.com; jintliu@sdu.edu.cn)More than 30 years of researches have confirmed that DES is a teratogen, which can cause malformations of an embryo or fetus. Prenatal exposure to DES is associated with an increased incidence of fibroids of adulthood [4,5]. Therefore, the Ministry of Agriculture of the People's Republic of China issued a banned list (Announcement No. 235; http://www. agri.gov.cn/ zcfg/bmgz/ t20060123_540865.htm, accessed on October 24, 2002) of veterinary drugs and announced that the use of DES in food and animal feed was prohibited. The European Union (EU) requires that residual levels of DES in edible animal food should not exceed 2 ng mL -1 [6]. Consequently, a rapid screening method with high sensitivity and

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“…The polyclonal OTC-BSA antibodies were generated locally and produced in New Zealand White rabbits based on the method developed by Zhang et al (2007). Isolation and purification of the polyclonal antibodies were conducted on a HiTrap Protein A column (Pharmacia) (Widiastuti et al, 2013).…”

Section: Elisa Developmentmentioning

confidence: 99%

Detection of Oxytetracycline in Broiler Chicken Meat Marketed in Several Cities in Java Island Using Enzyme-Linked Immunosorbent Assay (Elisa) Method

Widiastuti

Anastasia

2015

J. Indonesian Trop. Anim. Agric.

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ABSTRAKOksitetrasiklin (OTC) adalah salah satu jenis antibiotika golongan tetrasiklin berpektrum luas yang banyak digunakan pada industri ternak ayam. Namun penyalahgunaan dengan dosis berlebihan berpotensi terhadap terbentuknya residu pada produk ternak yang dapat membahayakan konsumen berupa reaksi alergis maupun resistensi. Penelitian ini bertujuan untuk mendeteksi residu OTC dalam daging ayam pedaging asal pasar tradisional dan swalayan di Depok-Bekasi, Bandung, Cilegon, Surakarta dan Yogyakarta secara indirect competitive enzyme-linked immunosorbent assay (icELISA). Hasil analisis pada 67 daging ayam pedaging menunjukkan hanya ada 1 (1,5%) sampel yang terdeteksi adanya residu OTC sebesar 86,1 ng/g dan masih di bawah batas maksimum residu OTC yang diijinkan (100 ng/g). Meskipun demikian, penerapan regulasi yang ketat terhadap penggunaan OTC ini di bidang peternakan ayam serta monitoring keberadaan residunya pada produk ternak sebelum dipasarkan sangat diperlukan untuk mencegah pengaruh buruk dari keberadaan residu tersebut pada produk pangan asal ternak. Kata kunci : oksitetrasiklin (OTC) residu, icELISA, daging ayam pedaging ABSTRACTOxytetracycline (OTC) is one of the tetracycline (TCs) broad-spectrum antibiotics widely used in the chicken industry. However, improper use of OTC with excessive doses potentially leads to residue formation in animal products that can be harmful to consumers in the form of allergic reaction or resistance. This study aimed to detect OTC residues in broiler chicken meat, marketed in traditional markets and supermarkets in Depok, Bekasi, Bandung, Cilegon, Surakarta and Yogyakarta using indirect competitive enzyme-linked immunosorbent assay (icELISA) method. The analyses of 67 broiler meat samples showed only 1 (1.5%) sample was positive for the OTC residue at 86.1 ng/g which meant below the maximum residue limits of permissible OTC (100 ng/g). Nevertheless, a stricter regulation for the use of OTC in the poultry industry and the monitoring of its residue in chicken products prior to marketing is still necessary to avoid the adverse effects of the residue present in animal products.

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Preparation of Anti-Tetracycline Antibodies and Development of an Indirect Heterologous Competitive Enzyme-Linked Immunosorbent Assay to Detect Residues of Tetracycline in Milk

Cited by 103 publications

References 25 publications

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Detection of Oxytetracycline in Broiler Chicken Meat Marketed in Several Cities in Java Island Using Enzyme-Linked Immunosorbent Assay (Elisa) Method

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