Sensitive lateral flow immunoassay of an antibiotic neomycin in foodstuffs


Aminoglycosides belong to a class of antibiotics now widely used in agriculture and veterinary medicine and expected to contaminate food products. In this study, a sensitive lateral flow immunoassay (LFIA) of an aminoglycoside neomycin (NEO) was developed. Two methods of immunochromatographic detection based on various techniques of gold nanoparticles (AuNPs) introduction as a label were compared. It was demonstrated that the indirect labeling (a conjugation of anti-species antibodies with a marker) allowed for an increase in assay sensitivity by 80 times. The test system was characterized by an instrumental limit of detection of 0.1 ng/mL and the cutoff level of 10 ng/mL; the assay duration was 15 min. Specificity only toward NEO was demonstrated. The developed LFIA has been tested to detect NEO in different foodstuffs. It has been demonstrated that 70–119% of NEO (coefficients of variations < 10%) can be determined in milk, turkey meat, honey, and eggs using simple procedures of preliminary sample preparation. Testing the samples showed the coincidence of the results for the developed lateral flow assay and for commercial ELISA kit.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Antibiotic Residue Test Kits Market (2019) Global industry perspective, comprehensive analysis and forecast, 2018–2025. Zion Market Research, New York, p 110

    Google Scholar 

  2. Bacanli M, Basaran N (2019) Importance of antibiotic residues in animal food. Food Chem Toxicol 125:462–466

    CAS  Article  Google Scholar 

  3. Berlina AN, Bartosh AV, Sotnikov DV, Zherdev AV, Xu C, Dzantiev BB (2018) Complexes of gold nanoparticles with antibodies in immunochromatography: comparison of direct and indirect immobilization of antibodies for the detection of antibiotics. Nanotechnol Russ 13(7):430–438

    CAS  Article  Google Scholar 

  4. Byzova NA, Safenkova IV, Slutskaya ES, Zherdev AV, Dzantiev BB (2017) Less is more: a comparison of antibody-gold nanoparticle conjugates of different ratios. Bioconjug Chem 28(11):2737–2746

    CAS  Article  Google Scholar 

  5. Byzova NA, Zvereva EA, Zherdev AV, Eremin SA, Dzantiev BB (2010) Rapid pretreatment-free immunochromatographic assay of chloramphenicol in milk. Talanta 81(3):843–848

    CAS  Article  Google Scholar 

  6. Byzova NA, Zvereva EA, Zherdev AV, Eremin SA, Sveshnikov PG, Dzantiev BB (2011) Pretreatment-free immunochromatographic assay for the detection of streptomycin and its application to the control of milk and dairy products. Anal Chim Acta 701(2):209–217

    CAS  Article  Google Scholar 

  7. Cháfer-Pericás C, Maquieira Á, Puchades R (2010) Fast screening methods to detect antibiotic residues in food samples. TrAC Trends Anal Chem 29(9):1038–1049

    Article  Google Scholar 

  8. Chen J, Ying GG, Deng WJ (2019) Antibiotic residues in food: extraction, analysis, and human health concerns. J Agric Food Chem 67(27):7569–7586

    CAS  Article  Google Scholar 

  9. Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nat Phys Sci 241(105):20–22

    CAS  Article  Google Scholar 

  10. Jaimee G, Halami PM (2016) Emerging resistance to aminoglycosides in lactic acid bacteria of food origin-an impending menace. Appl Microbiol Biotechnol 100(3):1137–1151

    CAS  Article  Google Scholar 

  11. Jin Y, Jang JW, Lee MH, Han CH (2006) Development of ELISA and immunochromatographic assay for the detection of neomycin. Clin Chim Acta 364(1–2):260–266

    CAS  Article  Google Scholar 

  12. Hendrickson OD, Zvereva EA, Shanin IA, Zherdev AV, Dzantiev BB (2019) Development of a multicomponent immunochromatographic test system for the detection of fluoroquinolone and amphenicol antibiotics in dairy products. J Sci Food Agric 99(8):3834–3842

    CAS  Article  Google Scholar 

  13. Hendrickson OD, Zvereva EA, Shanin IA, Zherdev AV, Tarannum N, Dzantiev BB (2018) Highly sensitive immunochromatographic detection of antibiotic ciprofloxacin in milk. Appl Biochem Microbiol 54(6):670–676

    CAS  Article  Google Scholar 

  14. House JR 3rd, House LK (2014) Ototoxicity of polymyxin B, neomycin, and hydrocortisone suspension in tympanoplasty surgery. Otolaryngol Head Neck Surg 150(2):282–284

    Article  Google Scholar 


  16. Lu Y, Sheng W, Liu B, Wang S (2017) ELISA-based sensing in food safety and quality analysis. In: Lu X (ed) Sensing techniques for food safety and quality control. Royal Society of Chemistry, Cambridge, pp 141–163

    Google Scholar 

  17. Parthasarathy R, Monette CE, Bracero S, S Saha M (2018) Methods for field measurement of antibiotic concentrations: limitations and outlook. FEMS Microbiol Ecol 94(8).

  18. Peng J, Wang Y, Liu L, Kuang H, Li A, Xu C (2016) Multiplex lateral flow immunoassay for five antibiotics detection based on gold nanoparticle aggregations. RSC Adv 6(10):7798–7805

    CAS  Article  Google Scholar 

  19. Perez-Rodriguez F, Mercanoglu Taban B (2019) A state-of-art review on multi-drug resistant pathogens in foods of animal origin: risk factors and mitigation strategies. Front Microbiol 10:2091

    Article  Google Scholar 

  20. Scott HM, Acuff G, Bergeron G, Bourassa MW, Gill J, Graham DW, Kahn LH, Morley PS, Salois MJ, Simjee S, Singer R, Smith TC, Storrs C, Wittum TE (2019) Critically important antibiotics: criteria and approaches for measuring and reducing their use in food animal agriculture. Ann N Y Acad Sci 1441(1):8–16

    Article  Google Scholar 

  21. Shi Q, Huang J, Sun Y, Deng R, Teng M, Li Q, Yang Y, Hu X, Zhang Z, Zhang G (2018a) A SERS-based multiple immuno-nanoprobe for ultrasensitive detection of neomycin and quinolone antibiotics via a lateral flow assay. Microchim Acta 85(2):84

    Article  Google Scholar 

  22. Shi Q, Huang J, Sun Y, Yin M, Hu M, Hu X, Zhang Z, Zhang G (2018b) Utilization of a lateral flow colloidal gold immunoassay strip based on surface-enhanced Raman spectroscopy for ultrasensitive detection of antibiotics in milk. Spectrochim Acta A Mol Biomol Spectrosc 197:107–113

    CAS  Article  Google Scholar 

  23. Thompson LA, Darwish WS (2019) Environmental chemical contaminants in food: review of a global problem. J Toxicol 2345283

  24. Uhrovcik J (2014) Strategy for determination of LOD and LOQ values–some basic aspects. Talanta 119:178–180

    CAS  Article  Google Scholar 

  25. Urusov AE, Petrakova AV, Zherdev AV, Dzantiev BB (2016) "Multistage in one touch" design with a universal labelling conjugate for high-sensitive lateral flow immunoassays. Biosens Bioelectr 86:575–579

    CAS  Article  Google Scholar 

  26. Urusov AE, Zherdev AV, Dzantiev BB (2014) Use of gold nanoparticle-labeled secondary antibodies to improve the sensitivity of an immunochromatographic assay for aflatoxin B1. Microchim Acta 181(15):1939–1946

    CAS  Article  Google Scholar 

  27. van Duijkeren E, Schwarz C, Bouchard D, Catry B, Pomba C, Baptiste KE, Moreno MA, Rantala M, Ružauskas M, Sanders P, Teale C, Wester AL, Ignate K, Kunsagi Z, Jukes H (2019) The use of aminoglycosides in animals within the EU: development of resistance in animals and possible impact on human and animal health: a review. J Antimicrob Chemother 74(9):2480–2496

    Article  Google Scholar 

  28. Wu Q, Peng D, Liu Q, Shabbir MAB, Sajid A, Liu Z, Wang Y, Yuan Z (2019) A novel microbiological method in microtiter plates for screening seven kinds of widely used antibiotics residues in milk, chicken egg and honey. Front Microbiol 10:436

    Article  Google Scholar 

  29. Xu F, Ren K, Yang Y-z, Guo J-p, Ma G-p, Liu Y-m, Lu Y-q, Li X-b (2015) Immunoassay of chemical contaminants in milk: a review. J Integr Agricult 14(11):2282–2295

    CAS  Article  Google Scholar 

  30. Yang B, Wang L, Luo C, Wang X, Sun C (2017) Simultaneous determination of 11 aminoglycoside residues in honey, milk, and pork by liquid chromatography with tandem mass spectrometry and molecularly imprinted polymer solid phase extraction. J AOAC Int 100(6):1869–1878

    CAS  Article  Google Scholar 

  31. Zheng J, Li Y, Guan H, Zhang J, Tan H (2019) Enhancement of neomycin production by engineering the entire biosynthetic gene cluster and feeding key precursors in Streptomyces fradiae CGMCC 4.576. Appl Microbiol Biotechnol 103(5):2263–2275.

    CAS  Article  Google Scholar 

Download references


The work was supported by the Russian Foundation for Basic Research, project 18-58-00038, and the Belarusian Republican Foundation for Fundamental Research, project X18P-060. The authors are grateful to S. M. Pridvorova (Federal Research Center "Fundamentals of Biotechnology") for obtaining electronic microphotographs of AuNPs and D. S. Popravko for developing Fig. 3.

Author information



Corresponding author

Correspondence to Boris B. Dzantiev.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interest.

Ethical approval

The studies did not involve human participants or animals.

Informed consent

Informed consent not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 186 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Hendrickson, O.D., Byzova, N.A., Zvereva, E.A. et al. Sensitive lateral flow immunoassay of an antibiotic neomycin in foodstuffs. J Food Sci Technol (2020).

Download citation


  • Aminoglycosides
  • Antibiotics
  • Food safety
  • Lateral flow assay
  • Neomycin