Microchimica Acta

, 186:448 | Cite as

Colorimetric aggregation assay for kanamycin using gold nanoparticles modified with hairpin DNA probes and hybridization chain reaction-assisted amplification

  • Chengnan Xu
  • Yibin Ying
  • Jianfeng PingEmail author
Original Paper


The authors describe a colorimetric method for determination of kanamycin by using gold nanoparticles (AuNPs) as the element of signal-conversion and by applying hybridization chain reaction-assisted signal amplification. The assay is carried out by monitoring the absorbance change and color change adding salt to the reaction solution containing kanamycin (analyte), hairpin DNA probe, and AuNPs. Three hairpin DNA probes with sticky ends were absorbed on the AuNPs via their sticky ends. Cating with DNA prevents them from salt-induced aggregation (which leads to a color change from red to blue) in the complete absence of kanamycin. In contrast, in the presence of kanamycin, the aptamer hairpin DNA probe binds kanamycin, and the newly exposed section of DNA triggers a cascade of hybridization chain reactions with formation of numerous dsDNAs. On addition of salt, the AuNPs form blue aggregates due to the repulsion between dsDNA and AuNPs. Under optimal conditions, the ration of absorbance at 520 and 630 nm drops with the kanamycin concentration in the range from 1 to 40 μM, and the limit of detection is 0.68 μM. The assay can selectively distinguish kanamycin from other antibiotics. The method was applied to kanamycin detection in (spiked) milk samples and gave excellent recoveries.

Graphical abstract

Schematic presentation of colorimetric method for kanamycin detection using gold nanoparticles modified with hairpin DNA probes and hybridization chain reaction-assisted amplification.


Kanamycin residue Colorimetric nanoprobe Hybridization chain reaction Gold nanoparticles Milk 



This research was supported by the National Natural Science Foundation of China (No. 31301468).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3574_MOESM1_ESM.docx (1.4 mb)
ESM 1 (DOCX 1438 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Biosystems Engineering and Food ScienceZhejiang UniversityHangzhouPeople’s Republic of China
  2. 2.Zhejiang A&F UniversityHangzhouPeople’s Republic of China

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