Food Analytical Methods

, Volume 12, Issue 11, pp 2430–2437 | Cite as

An Immunochromatographic Lateral Flow Strip Test for the Rapid Detection of Danofloxacin in Milk

  • Xingdong Yang
  • Yinbiao Wang
  • Jifei Yang
  • Zhongke Sun
  • Zonghao Yue
  • Lili Li
  • Le He
  • Xiaofei HuEmail author


An immunochromatographic lateral flow strip test (ILFST) for the rapid detection of danofloxacin (DAN) in milk samples was developed based on colloidal gold-labeled anti-DAN monoclonal antibody 3E9 and a competitive format. Quantitative detections of DAN-spiked samples using the ILFST gave a half inhibitory concentration (IC50) of 0.513 ng/mL and a limit of detection (LOD) of 0.092 ng/mL. Recoveries of DAN from spiked milk samples were from 80.0 to 95.1% within an assay (intra-assay) and from 87.0 to 89.8% between assays (inter-assay). The coefficients of variation (CV) for intra-assay and inter-assay were 2.09–9.93% and 2.42–12.07%, respectively. These data indicated that the ILFST was highly sensitive and reproducible. Detections of authentic milk samples using ILFST and high-performance liquid chromatography (HPLC) showed no significant differences between the two methods. Hence, the ILFST can be widely used for both qualitative and quantitative detection of DAN residues in milk samples.


Danofloxacin Monoclonal antibody Colloidal gold Immunochromatographic lateral flow strip test Rapid and quantitative test 


Funding Information

This study was supported by the National Science & Technology Pillar Program of the 12th Five-Year Plan (Grant No. 2014BAD13B05), the Natural Science Foundation of Henan Province (Grant No. 182300410093), and Henan Province Science and Technology Agency (Grant No. 182102310683).

Compliance with Ethical Standards

Conflict of Interest

Xingdong Yang declares that he has no conflict of interest. Yinbiao Wang declares that he has no conflict of interest. Jifei Yang declares that he has no conflict of interest. Zhongke Sun Wang declares that he has no conflict of interest. Zonghao Yue declares that he has no conflict of interest. Lili Li declares that she has no conflict of interest. Le He declares that he has no conflict of interest. Xiaofei Hu declares that he has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants performed by any of the authors. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed consent

Not applicable.


  1. Bucknall S, Silverlight J, Coldham N, Thorne L, Jackman R (2003) Antibodies to the quinolones and fluoroquinolones for the development of generic and specific immunoassays for detection of these residues in animal products. Food Addit Contam 20:221–228CrossRefGoogle Scholar
  2. Chen XL, Xu HY, Lai WH, Chen Y, Yang XH, Xiong YH (2012) A sensitive chromatographic strip test for the rapid detection of enrofloxacin in chicken muscle. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 29:383–391PubMedGoogle Scholar
  3. Chen M, Wen K, Tao XQ, Ding SY, Xie J, Yu XZ, Li JC, Xia X, Wang Y, Xie SL, Jiang HY (2014) A novel multiplexed fluorescence polarisation immunoassay based on a recombinant bi-specific single-chain diabody for simultaneous detection of fluoroquinolones and sulfonamides in milk. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 31:1959–1967CrossRefGoogle Scholar
  4. Dimitrova DJ, Haritova AM, Dinev TD, Moutafchieva RG, Lashev LD (2014) Comparative pharmacokinetics of danofloxacin in common pheasants, guinea fowls and Japanese quails after intravenous and oral administration. Br Poult Sci 55:120–125CrossRefGoogle Scholar
  5. Drlica K (1999) Mechanism of fluoroquinolone action. Curr Opin Microbiol 2:504–508CrossRefGoogle Scholar
  6. Drlica K, Zhao X (1997) DNA gyrase, topoisomerase IV, and the 4-quinolones. Microbiol Mol Biol Rev 61:388–392Google Scholar
  7. Duan JH, Yuan ZH (2001) Development of an indirect competitive ELISA for ciprofloxacin residues in food animal edible tissues. J Agric Food Chem 49:1087–1089CrossRefGoogle Scholar
  8. Holtzapple CK, Buckley SA, Stanker LH (1997) Production and characterization of monoclonal antibodies against sarafloxacin and cross-reactivity studies of related fluoroquinolones. J Agric Food Chem 45:1984–1990CrossRefGoogle Scholar
  9. Huet AC, Charlier C, Tittlemier SA, Singh G, Benrejeb S, Delahaut P (2006) Simultaneous determination of (fluoro)quinolone antibiotics in kidney, marine products, eggs, and muscle by enzyme-linked immunosorbent assay (ELISA). J Agric Food Chem 54:2822–2827CrossRefGoogle Scholar
  10. Inc P (1988) Nonclinical total residue depletion study in broiler chickens. Unpublished Study No. 1515N-60-88-006 from Pfizer Central Research, Terre Haute, IN and Groton, CTGoogle Scholar
  11. Inc P (1995) Determination of danofloxacin and N-desmethyl in cattle liver. Cross validation in cattle muscle, kidney, and fat. Cross validation in chicken liver and muscle. Unpublished Study No. DM95-Advocin-001 from Pfizer Central Research, Groton, CTGoogle Scholar
  12. Jacoby GA (2005) Mechanisms of Resistance to Quinolones. Clin Infect Dis 41, S:120-126.CrossRefGoogle Scholar
  13. Jiang WX, Wang ZH, Beier RC, Jiang HY, Wu YN, Shen JZ (2013) Simultaneous determination of 13 fluoroquinolone and 22 sulfonamide residues in milk by a dual-colorimetric enzyme-linked immunosorbent assay. Anal Chem 85:1995–1999CrossRefGoogle Scholar
  14. Klein U, de Jong A, Moyaert H, EI Garch F, Leon R, Richard-Mazet A, Rose M, Maes D, Pridmore A, Thomson JR, Ayling RD (2017) Antimicrobial susceptibility monitoring of Mycoplasma hyopneumoniae and Mycoplasma bovis isolated in Europe. Vet Microbiol 204:188–193CrossRefGoogle Scholar
  15. Kong DZ, Wu XL, Li Y, Liu LQ, Song SS, Zheng QK, Kuang H, Xu CL (2019) Ultrasensitive and eco-friendly immunoassays based monoclonal antibody for detection of deoxynivalenol in cereal and feed samples. Food Chem 270:130–137CrossRefGoogle Scholar
  16. Li XW, Zhang GP, Liu QT, Feng CH, Wang XN, Yang YY, Xiao ZJ, Yang JF, Xing GX, Zhao D, Cai SJ, Chen HC (2009) Development of immunoassays for the detection of sulfamethazine in swine urine. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 26:314–325CrossRefGoogle Scholar
  17. Licata A, Randazzo C, Morreale I, Butera G, D’Alessandro N, Craxì A (2012) Fluoroquinolone-induced liver injury: three new cases and a review of the literature. Eur J Clin Pharmacol 68:525–532CrossRefGoogle Scholar
  18. Liu LQ, Luo LJ, Suryoprabowo S, Peng J, Kuang H, Xu CL (2014) Development of an immunochromatographic strip test for rapid detection of ciprofloxacin in milk samples. Sensors (Basel) 14:16785–16798CrossRefGoogle Scholar
  19. Martinez M, McDermott P, Walker R (2006) Pharmacology of the fluoroquinolones: a perspective for the use in domestic animals. Vet J 172:10–28CrossRefGoogle Scholar
  20. Moreno-González D, Lara FJ, Gámiz-Gracia L, García-Campaña AM (2014) Molecularly imprinted polymer as in-line concentrator in capillary electrophoresis coupled with mass spectrometry for the determination of quinolones in bovine milk samples. J Chromatogr A 1360:1–8CrossRefGoogle Scholar
  21. Petrakova AV, Urusova AE, Voznyak MV, Zherdeva AV, Dzantiev BB (2015) Immunochromatographic test system for the detection of T-2 toxin. Prikl Biokhim Mikrobiol 51:616–623PubMedGoogle Scholar
  22. Rusenova N, Haritova AM, Parvanov PR (2014) Influence of cyclosporine A and quercetine on MIC values of danofloxacin mesylate in Escherichia coli strains isolated from poultry. Turk J Vet Anim Sci 33:247–251Google Scholar
  23. Sarasola P, Lees P, AliAbadi FS, McKellar QA, Donachie W, Marr KA, Sunderland SJ, Rowan TG (2002) Pharmacokinetic and pharmacodynamic profiles of danofloxacin administered by two dosing regimens in calves infected with mannheimia (Pasteurella) haemolytica. Antimicrob Agents Chemother 46:3013–3019CrossRefGoogle Scholar
  24. Shim WB, Kim JS, Kim MG, Chung DH (2013) Rapid and sensitive immunochromatographic strip for on-site detection of sulfamethazine in meats and eggs. J Food Sci 78:M1575–M1581CrossRefGoogle Scholar
  25. Suryoprabowo S, Liu LQ, Peng J, Kuang H, Xu CL (2014) Development of a broad specific monoclonal antibody for fluoroquinolone analysis. Food Anal. Methods 7:2163–2168CrossRefGoogle Scholar
  26. Tittlemier SA, Gélinas J-M, Dufresne G, Haria M, Querry J, Cleroux C, Ménard C, Delahaut P, Singh G, Fischer-Durand N, Godefroy SB (2008) Development of a direct competitive enzyme-linked immunosorbent assay for the detection of fluoroquinolone residues in shrimp. Food Anal. Methods 1:28–35CrossRefGoogle Scholar
  27. Tufa RA, Pinacho DG, Pascual N, Granados M, Companyó R, Marco MP (2015) Development and validation of an enzyme linked immunosorbent assay for fluoroquinolones in animal feeds. Food Control 57:195–201CrossRefGoogle Scholar
  28. Wang ZH, Zhu Y, Ding SY, He FY, Beier RC, Li JC, Jiang HY, Feng CW, Wan YP, Zhang SX, Kai ZP, Yang XL, Shen JZ (2007) Development of a monoclonal antibody-based broad-specificity ELISA for fluoroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds. Anal Chem 79:4471–4483CrossRefGoogle Scholar
  29. Wang Y, Li ZZ, Pei YF, Li QM, Sun YN, Yang JF, Yang YY, Zhi YB, Deng RG, Hou YZ, Hu XF (2017) Establishment of a lateral flow colloidal gold immunoassay strip for the rapid detection of soybean allergen β-conglycinin. Food Anal Methods 10:2429–2435CrossRefGoogle Scholar
  30. Wassenaar TM (2008) Use of antimicrobial agents in veterinary medicine and implications for human health. Crit Rev Microbiol 31:155–169CrossRefGoogle Scholar
  31. Watanabe H, Satake A, Kido Y, Tsuji A (2002) Monoclonal-based enzyme-linked immunosorbent assay and immunochromatographic assay for enrofloxacin in biological matrices. Analyst. 127:98–103CrossRefGoogle Scholar
  32. Yang XD, Wang FY, Song CM, Wu SY, Zhang GP, Zeng XY (2015a) Establishment of a lateral flow colloidal gold immunoassay strip for the rapid detection of estradiol in milk samples. LWT Food Sci Technol 64:88–94CrossRefGoogle Scholar
  33. Yang XD, Zhang GP, Wang FY, Wang YB, Hu XF, Li QM, Jia GC, Liu Z, Wang Y,Deng, RG, Zeng XY (2015b) Development of a colloidal gold-based strip test for the detection of chlorothalonil residues in cucumber. Food Agr Immunol 26:729–737CrossRefGoogle Scholar
  34. Yang MM, Zhao YT, Wang LM, Paulsen M, Simpson CD, Liu FQ, Du D, Lin YH (2018) Simultaneous detection of dual biomarkers from humans exposed to organophosphorus pesticides by combination of immunochromatographic test strip and ellman assay. Biosens Bioelectron 104:39–44CrossRefGoogle Scholar
  35. Zhang GP, Guo JQ, Wang XN, Yang JX, Yang YY, Li QM, Li XW, Deng RG, Xiao ZJ, Yang JF et al (2006) Development and evaluation of an immuno-chromatographic strip for trichinellosis detection. Vet Parasitol 137:286–293CrossRefGoogle Scholar
  36. Zhang GP, Guo JQ, Wang XN (2009) Immunochromatographic lateral flow strip tests. Methods Mol Biol 504:169–183CrossRefGoogle Scholar
  37. Zhang XD, Wang CC, Yang LY, Zhang W, Lin J, Li C (2017) Determination of eight quinolones in milk using immunoaffinity microextraction in a packed syringe and liquid chromatography with fluorescence detection. J Chromatogr B Anal Technol Biomed Life Sci 1064:68–74CrossRefGoogle Scholar
  38. Zhao YL, Zhang GP, Liu QT, Teng M, Yang J, Wang JH (2008) Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of enrofloxacin residues. J Agric Food Chem 56:12138–12142CrossRefGoogle Scholar
  39. Zhi AM, Li BB, Liu QT, Hu XF, Zhao D, Hu YZ, Deng RG, Chai SJ, Zhang GP (2010) Development of a lateral-flow immunochromatographic test device for the rapid detection of difloxacin residues. Food Agric Immunol 21:335–345CrossRefGoogle Scholar
  40. Zhu Y, Li L, Wang ZH, Chen YQ, Zhao ZM, Zhu L, Wu XP, Wan YP, He FY, Shen JZ (2008) Development of an immunochromatography strip for the rapid detection of 12 fluoroquinolones in chicken muscle and liver. J Agric Food Chem 56:5469–5474CrossRefGoogle Scholar
  41. Ziarrusta H, Val N, Dominguez H, Mijangos L, Prieto A, Usobiaga A, Etxebarria N, Zuloaga O, Olivares M (2017) Determination of fluoroquinolones in fish tissues, biological fluids, and environmental waters by liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 409:6359–6370CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Food and Drug InspectionZhoukou Normal UniversityZhoukouPeople’s Republic of China
  2. 2.Key Laboratory of Animal Immunology of the Ministry of Agriculture, Henan Provincial Key Laboratory of Animal ImmunologyHenan Academy of Agricultural SciencesZhengzhouPeople’s Republic of China
  3. 3.School of Public HealthXinxiang Medical UniversityXinxiangPeople’s Republic of China

Personalised recommendations