Microchimica Acta

, 186:82 | Cite as

A lateral flow assay for copper(II) utilizing catalytic and stem-loop based signal amplification

  • Yulong Wang
  • Limin Wang
  • Cunzheng ZhangEmail author
  • Fengquan LiuEmail author
Original Paper


A DNAzyme-based catalytic and stem-loop based amplification scheme is used in a Cu(II)-specific lateral flow assay (LFA). Three test lines with given cut-off value on the test strip are set as the signal indicating zone for semiquantitative analysis by the number of red color bands that appear after lateral flow. The colored bands are generated by accumulation of gold nanoparticles. Four detection ranges can be visualied: (a) 0–2 ng·mL−1 (= negative); 2–50 ng·mL−1; 50–200 ng·mL−1 and > 200 ng·mL−1 of Cu(II) (= positive). The visual detection limit is thus considered as being 2 ng·mL−1 which is much lower than the U.S. EPA limit in drinking water (1.25 μg·mL−1). The highly specific DNAzyme, the strong multiple-turnover catalytic target recycling property and highly efficient amplification strategy warrant the high specificity, sensitivity and rapidity of this LFA. Conceivbly, this detecton scheme can be extended to other metal ions by proper choice of the ion-specific DNAzyme.

Graphical abstract

Schematic presentation of a semiquantitative lateral flow test strip for Cu2+ analysis by three visual cut-off value test lines using catalytic and stem-loop based signal amplification strategy.


Target recycling DNAzyme Gold nanoparticle Semiquantitative Visual detection Copper ions 



This work was supported by the earmarked fund for China Agriculture Research System (CARS-28-16), the Ministry of Science and Technology of People’s Republic of China (2016YFD0200803-3), the Department of Finance of Jiangsu Province [CX(17)1003] and the Department of Science and Technology of Jiangsu Province (BE2014722).

Compliance with ethical standards

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

Supplementary material

604_2018_3197_MOESM1_ESM.doc (140 kb)
ESM 1 (DOC 139 kb)


  1. 1.
    Xie QY, Wu YH, Xiong QR, Xu HY, Xiong YH, Liu K, Jin Y, Lai WH (2014) Advantages of fluorescent microspheres compared with colloidal gold as a label in immunochromatographic lateral flow assays. Biosens Bioelectron 54:262–265CrossRefGoogle Scholar
  2. 2.
    Quesada-González D, Merkoçi A (2015) Nanoparticle-based lateral flow biosensors. Biosens Bioelectron 73:47–63CrossRefGoogle Scholar
  3. 3.
    Bruno JG (2014) Application of DNA aptamers and quantum dots to lateral flow test strips for detection of foodborne pathogens with improved sensitivity versus colloidal gold. Pathogens 3(2):341–355CrossRefGoogle Scholar
  4. 4.
    Chen AL, Yang SM (2015) Replacing antibodies with aptamers in lateral flow immunoassay. Biosens Bioelectron 71:230–242CrossRefGoogle Scholar
  5. 5.
    Anfossi L, Baggiani C, Giovannoli C, Biagioli F, D’Arco G, Giraudi G (2013) Optimization of a lateral flow immunoassay for the ultrasensitive detection of aflatoxin M1 in milk. Anal Chim Acta 772:75–80CrossRefGoogle Scholar
  6. 6.
    Berlina AN, Taranova NA, Zherdev AV, Vengerov YY, Dzantiev BB (2013) Quantum dot-based lateral flow immunoassay for detection of chloramphenicol in milk. Anal Bioanal Chem 405:4997–5000CrossRefGoogle Scholar
  7. 7.
    Abera A, Choi JW (2010) Quantitative lateral flow immunosensor using carbon nanotubes as label. Anal Methods 2:1819–1822CrossRefGoogle Scholar
  8. 8.
    Mao X, Wang W, Du TE (2013) Dry-reagent nucleic acid biosensor based on blue dye doped latex beads and lateral flow strip. Talanta 114:248–253CrossRefGoogle Scholar
  9. 9.
    Corstjens PL, van Lieshout L, Zuiderwijk M, Kornelis D, Tanke HJ, Deelder AM, van Dam GJ (2008) Up-converting phosphor technology-based lateral flow assay for detection of Schistosoma circulating anodic antigen in serum. J Clin Microbiol 46:171–176Google Scholar
  10. 10.
    Liu X, Xiang JJ, Tang Y, Zhang XL, Fu QQ, Zou JH, Lin YH (2012) Colloidal gold nanoparticle probe-based immunochromatographic assay for the rapid detection of chromium ions in water and serum samples. Anal Chim Acta 745:99–105CrossRefGoogle Scholar
  11. 11.
    Park JM, Jung HW, Chang YW, Kim HS, Kang MJ, Pyun JC (2015) Chemiluminescence lateral flow immunoassay based on Pt nanoparticle with peroxidase activity. Anal Chim Acta 853:360–367CrossRefGoogle Scholar
  12. 12.
    Chen ZH, Liang RL, Guo XX, Liang JY, Deng QT, Li M, An TX, Liu TC, Wu YS (2017) Simultaneous quantitation of cytokeratin-19 fragment and carcinoembryonic antigen in human serum via quantum dot-doped nanoparticles. Biosens Bioelectron 91:60–65CrossRefGoogle Scholar
  13. 13.
    Murray CB, Kagan CR, Bawendi MG (2000) Synthesis and characterization of monodisperse nanocrystals and close-packed nanocrystal assemblies. Annu Rev Mater Sci 30:545–610CrossRefGoogle Scholar
  14. 14.
    He YQ, Zeng K, Zhang SQ, Gurung AS, Baloda M, Zhang XB, Liu GD (2012) Visual detection of gene mutations based on isothermal strand-displacement polymerase reaction and lateral flow strip. Biosens Bioelectron 31:310–315CrossRefGoogle Scholar
  15. 15.
    Chen JH, Zhou SG, Wen JL (2014) Disposable strip biosensor for visual detection of Hg2+ based on Hg2+-triggered toehold binding and exonuclease III-assisted signal amplification. Anal Chem 86:3108–3114CrossRefGoogle Scholar
  16. 16.
    Rosser A, Rollinson D, Forrest M, Webster BL (2015) Isothermal recombinase polymerase amplification (RPA) of Schistosoma haematobium DNA and oligochromatographic lateral flow detection. Parasit Vectors 8:446CrossRefGoogle Scholar
  17. 17.
    Huang Y, Wang WQ, Wu TT, Xu LP, Wen YQ, Zhang XJ (2016) A three-line lateral flow biosensor for logic detection of microRNA based on Y-shaped junction DNA and target recycling amplification. Anal Bioanal Chem 408:8195–8202CrossRefGoogle Scholar
  18. 18.
    Foo PC, Chan YY, Mohamed M, Wong WK, Najian ABN, Lim BH (2017) Development of a thermostabilised triplex LAMP assay with dry-reagent four target lateral flow dipstick for detection of Entamoeba histolytica and non-pathogenic Entamoeba spp. Anal Chim Acta 966:71–80CrossRefGoogle Scholar
  19. 19.
    Jauset-Rubio M, Svobodová M, Mairal T, McNeil C, Keegan N, Saeed A, Abbas MN, EI-Shahawi MS, Bashammakh AS, Alyoubi AO, O Sullivan CK (2017) Ultrasensitive, rapid and inexpensive detection of DNA using paper based lateral flow assay. Sci Rep 6:37732Google Scholar
  20. 20.
    Wang ZD, Lee JH, Lu Y (2008) Label-free colorimetric detection of Lead ions with a Nanomolar detection limit and tunable dynamic range by using gold nanoparticles and DNAzyme. Adv Mater 20:3263–3267CrossRefGoogle Scholar
  21. 21.
    Liu JW, Lu Y (2007) Rational design of “turn-on” allosteric DNAzyme catalytic beacons for aqueous mercury ions with ultrahigh sensitivity and selectivity. Angew Chem Int Ed 119:7731–7734CrossRefGoogle Scholar
  22. 22.
    Liu JW, Lu Y (2003) A colorimetric lead biosensor using DNAzyme-directed assembly of gold nanoparticles. J Am Chem Soc 125:6642–6643CrossRefGoogle Scholar
  23. 23.
    Hollenstein M, Hipolito C, Lam C, Dietrich D, Perrin DM (2008) A highly selective DNAzyme sensor for mercuric ions. Angew Chem Int Ed 47:4346–4350CrossRefGoogle Scholar
  24. 24.
    Yang Y, Yuan Z, Liu XP, Liu Q, Mao CJ, Niu HL, Jin BK, Zhang SY (2016) Electrochemical biosensor for Ni2+ detection based on a DNAzyme-CdSe nanocomposite. Biosens Bioelectron 77:13–18CrossRefGoogle Scholar
  25. 25.
    Zhang XB, Wang ZD, Xing H, Xiang Y, Lu Y (2010) Catalytic and molecular beacons for amplified detection of metal ions and organic molecules with high sensitivity. Anal Chem 82:5005–5011CrossRefGoogle Scholar
  26. 26.
    Zhao XH, Kong RM, Zhang XB, Meng HM, Liu WN, Tan WH, Shen GL, Yu RQ (2011) Graphene-DNAzyme based biosensor for amplified fluorescence “TurnOn” detection of Pb2+ with a high selectivity. Anal Chem 83:5062–5066CrossRefGoogle Scholar
  27. 27.
    Zhang XB, Kong RM, Lu Y (2011) Metal ion sensors based on DNAzymes and related DNA molecules. Annu Rev Anal Chem 4:105–128CrossRefGoogle Scholar
  28. 28.
    Hahn SH, Tanner MS, Danke DM, Gahl WA (1995) Normal metallothionein synthesis in fibroblasts obtained from children with Indian childhood cirrhosis or copper-associated childhood cirrhosis. Biochem Mol Med 54:142–145CrossRefGoogle Scholar
  29. 29.
    Brewer GJ (2009) The risks of copper toxicity contributing to cognitive decline in the aging population and to Alzheimer's disease. J Am Coll Nutr 28:238–242CrossRefGoogle Scholar
  30. 30.
    Gao Y, Deng XL, Wen W, Zhang XH, Wang SF (2017) Ultrasensitive paper based nucleic acid detection realized by threedimensional DNA-AuNPs network amplification. Biosens Bioelectron 92:529–535CrossRefGoogle Scholar
  31. 31.
    Wang YL, Wang LM, Xue JJ, Dong JB, Cai J, Hua XD, Wang MH, Zhang CZ, Liu FQ (2017) Signal-amplified lateral flow test strip for visual detection of Cu2+. PLoS One 12(1):e0169345CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base, Key Laboratory of Control Technology and Standard for Agro-product Safety and Quality, Ministry of AgricultureJiangsu Academy of Agricultural SciencesNanjingPeople’s Republic of China
  2. 2.Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and TechnologyInstitute of Plant Protection, Jiangsu Academy of Agricultural SciencesNanjingChina
  3. 3.College of Plant Protection (Key Laboratory of Integrated Management of Crop Diseases and Pests)Nanjing Agricultural UniversityNanjingPeople’s Republic of China

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