Advertisement

Microchimica Acta

, Volume 183, Issue 10, pp 2791–2797 | Cite as

Aptamer based fluorescent cocaine assay based on the use of graphene oxide and exonuclease III-assisted signal amplification

  • Yulin Zhang
  • Zhongyue Sun
  • Lina Tang
  • Hong ZhangEmail author
  • Guo-Jun ZhangEmail author
Original Paper

Abstract

The article reports an aptamer based assay for cocaine by employing graphene oxide and exonuclease III-assisted signal amplification. It is based on the following scheme and experimental steps: (1) Exo III can digest dsDNA with blunt or recessed 3-terminus, but it has limited activity to ssDNA or dsDNA with protruding 3-terminus; (2) GO can absorb the FAM-labeled ssDNA probe and quench the fluorescence of probe, while the affinity between FAM-labeled mononucleotide and GO is negligible; (3) Cocaine aptamer can be split into two flexible ssDNA pieces (Probe 1 and Probe 2) without significant perturbation of cocaine-binding abilities; (4) The triple complex consisting of Probe 1, Probe 2 and cocaine can be digested by Exo III with the similar efficiency as normal dsDNA. Cocaine aptamer is split into two flexible ssDNA pieces (Probe 2 and 3′-FAM-labeled Probe 1). Cocaine can mediate the cocaine aptamer fragments forming a triplex. The triple complex has unique characteristic with 3′-FAM-labeled blunt end at the Probe 1 and 3′-overhang end at Probe 2. If exonuclease III is added, it will catalyze the stepwise removal of fluorescein (FAM) labeled mononucleotides from the 3-hydroxy termini of the special triplex complex, resulting in liberation of cocaine. The cocaine released in this step can produce a new cleavage cycle, thereby leading to target recycling. Through such a cyclic bound-hydrolysis process, small amounts of cocaine can induce the cleavage of a large number of FAM-labeled probe 1. The cleaved FAM-labeled mononucleotides are not adsorbed on the surface of graphene oxide (GO), so a strong fluorescence signal enhancement is observed as the cocaine triggers enzymatic digestion. Under optimized conditions, the assay allows cocaine to be detected in the 1 to 500 nM concentration range with a detection limit of 0.1 nM. The method was applied to the determination of cocaine in spiked human plasma, with recoveries ranging from 92.0 to 111.8 % and RSD of <12.8 %.

Graphical abstract

Aptamer based fluorescent cocaine assay based on graphene oxide and exonuclease III-assisted signal amplification

Keywords

Drug analysis Cocaine aptamer fragment Exo III triggered amplification Enzymatic digestion Plasma analysis 

Notes

Acknowledgments

The authors acknowledge the support of Natural Science Foundation of China (Nos. 21275040 and 21475034).

Compliance with ethical standards

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

Supplementary material

604_2016_1923_MOESM1_ESM.doc (1008 kb)
ESM 1 (DOC .98 mb)

References

  1. 1.
    Xie SJ, Zhou H, Liu D-Y, Shen G-L, R-Q Y, Z-S W (2013) In situ amplification signaling-based autonomous aptameric machine for the sensitive fluorescence detection of cocaine. Biosens Bioelectron 44:95–100CrossRefGoogle Scholar
  2. 2.
    Jiang B, Wang M, Chen Y, Xie J, Xiang Y (2012) Highly sensitive electrochemical detection of cocaine on graphene/aunp modified electrode via catalytic redox-recycling amplification. Biosens Bioelectron 32:305–308CrossRefGoogle Scholar
  3. 3.
    Zhou J, Ellis A-V, Kobus H, Voelcker N-H (2012) Aptamer sensor for cocaine using minor groove binder based energy transfer. Anal Chim Acta 719:76–81CrossRefGoogle Scholar
  4. 4.
    Stojanovic M-N, de Prada P, Landry D-W (2000) Fluorescent sensors based on aptamer self-assembly. J Am Chem Soc 122:11547–11548CrossRefGoogle Scholar
  5. 5.
    Chen Z, Tan Y, Xu K, Zhang L, Qiu B, Guo L, Lin Z, Chen G (2016) Stimulus-response mesoporous silica nanoparticle-based chemiluminescence biosensor for cocaine determination. Biosens Bioelectron 75:8–14CrossRefGoogle Scholar
  6. 6.
    Taghdisi S-M, Danesh N-M, Emrani A-S, Ramezani M, Abnous K (2015) A novel electrochemical aptasensor based on single-walled carbon nanotubes, gold electrode and complimentary strand of aptamer for ultrasensitive detection of cocaine. Biosens Bioelectron 73:245–250CrossRefGoogle Scholar
  7. 7.
    Mao Y, Chen Y, Li S, Lin S, Jiang Y (2015) A graphene-based biosensing platform based on regulated release of an aptameric DNA biosensor. Sensors 15:28244CrossRefGoogle Scholar
  8. 8.
    Soh J-H, Lin Y, Rana S, Ying J-Y, Stevens MM (2015) Colorimetric detection of small molecules in complex matrixes via target-mediated growth of aptamer-functionalized gold nanoparticles. Anal Chem 87:7644–7652CrossRefGoogle Scholar
  9. 9.
    Ma C, Wang W, Yang Q, Shi C, Cao L (2011) Cocaine detection via rolling circle amplification of short DNA strand separated by magnetic beads. Biosens Bioelectron 26:3309–3312CrossRefGoogle Scholar
  10. 10.
    Huang J, Chen Y, Yang L, Zhu Z, Zhu G, Yang X, Wang K, Tan W (2011) Amplified detection of cocaine based on strand-displacement polymerization and fluorescence resonance energy transfer. Biosens Bioelectron 28:450–453CrossRefGoogle Scholar
  11. 11.
    Zhang H, Hu X, Fu X (2014) Aptamer-based microfluidic beads array sensor for simultaneous detection of multiple analytes employing multienzyme-linked nanoparticle amplification and quantum dots labels. Biosens Bioelectron 57:22–29CrossRefGoogle Scholar
  12. 12.
    Zhang K, Wang K, Zhu X, Zhang J, Xu L, Huang B, Xie M (2014) Label-free and ultrasensitive fluorescence detection of cocaine based on a strategy that utilizes DNA-templated silver nanoclusters and the nicking endonuclease-assisted signal amplification method. Chem Commun 50:180–182CrossRefGoogle Scholar
  13. 13.
    Xie S-J, Zhou H, Liu D, Shen G-L, Yu R, Z-S W (2013) In situ amplification signaling-based autonomous aptameric machine for the sensitive fluorescence detection ofcocaine. Biosens Bioelectron 44:95–100CrossRefGoogle Scholar
  14. 14.
    Zhang Y-L, Tang LN, Yang F, Sun Z-Y, Zhang G-J (2015) Highly sensitive DNA-based fluorometric mercury(ii) bioassay based on graphene oxide and exonuclease iii-assisted signal amplification. Microchim Acta 182:1535–1541CrossRefGoogle Scholar
  15. 15.
    Chen H-G, Ren W, Jia J, Feng J, Gao Z-F, Li N-B, Luo H-Q (2016) Fluorometric detection of mutant DNA oligonucleotide based on toehold strand displacement-driving target recycling strategy and exonuclease iii-assisted suppression. Biosens Bioelectron 77:40–45CrossRefGoogle Scholar
  16. 16.
    Xu Q, Cao A, Zhang L-f, Zhang C-y (2012) Rapid and label-free monitoring of exonuclease iii-assisted target recycling amplification. Anal Chem 84:10845–10851CrossRefGoogle Scholar
  17. 17.
    Huang K-J, Shuai H-L, Zhang J-Z (2016) Ultrasensitive sensing platform for platelet-derived growth factor bb detection based on layered molybdenum selenide–graphene composites and exonuclease iii assisted signal amplification. Biosens Bioelectron 77:69–75CrossRefGoogle Scholar
  18. 18.
    Jalaja K, Sreehari VS, Kumar PRA, Nirmala RJ (2016) Graphene oxide decorated electrospun gelatin nanofibers: fabrication, properties and applications. Mater Sci Eng C 64:11–19CrossRefGoogle Scholar
  19. 19.
    Liu X, Wang J-Y, Mao X-B, Ning Y, Zhang G-J (2015) Single-shot analytical assay based on graphene-oxide-modified surface acoustic wave biosensor for detection of single-nucleotide polymorphisms. Anal Chem 87:9352–9359CrossRefGoogle Scholar
  20. 20.
    Cai BJ, Huang L, Zhang H, Sun Z-Y, Zhang Z-Y, Zhang G-J (2015) Gold nanoparticles-decorated graphene field-effect transistor biosensor for femtomolar microrna detection. Biosens Bioelectron 74:329–334CrossRefGoogle Scholar
  21. 21.
    Lei Y, Yang F, Tang L-N, Chen K-L, Zhang G-J (2015) Identification of chinese herbs using a sequencing-free nanostructured electrochemical DNA biosensor. Sensors 15:29882–29892CrossRefGoogle Scholar
  22. 22.
    Guo S, Yang F, Zhang Y-L, Ning Y, Yao Q-F, Zhang G-J (2014) Amplified fluorescence sensing of mirna by combination of graphene oxide with duplex-specific nuclease. Anal Methods 6:3598–3603CrossRefGoogle Scholar
  23. 23.
    Shi Y, Dai H-C, Sun Y-J, Hu J-T, Ni P-J, Li Z (2013) Fluorescent sensing of cocaine based on a structure switching aptamer, gold nanoparticles and graphene oxide. Analyst 138:7152–7156CrossRefGoogle Scholar
  24. 24.
    Qiu L, Zhou H, Zhu W-P, Qiu L-P, Jiang J-H, Shen G-L, R-Q Y (2013) A novel label-free fluorescence aptamer-based sensor method for cocaine detection based on isothermal circular strand-displacement amplification and graphene oxide absorption. New J Chem 37:3998–4003CrossRefGoogle Scholar
  25. 25.
    Chen J, Zeng L (2013) Enzyme-amplified electronic logic gates based on split/intactaptamers. Biosens Bioelectron 42:93–99CrossRefGoogle Scholar
  26. 26.
    He Y, Jiao B, Tang H (2014) Interaction of single-stranded DNA with graphene oxide: fluorescence study and its application for S1 nuclease detection. RSC Adv 35:18294–18300CrossRefGoogle Scholar
  27. 27.
    Zhao X-H, Kong R-M, Zhang X-B, Meng H-M, Liu W-N, Tan W, Shen G-L, R-Q Y (2011) Graphene–DNAzyme based biosensor for amplified fluorescence “turn-on” detection of Pb2+ with a high selectivity. Anal Chem 83:5062–5066CrossRefGoogle Scholar
  28. 28.
    Niu S-Y, Lou X-F, Jiang Y, Lin J-H (2012) A novel fluorescence sensor for cocaine with signal amplification through cycling exo-cleaving with a hairpin probe. Anal Lett 45:1919–1927CrossRefGoogle Scholar
  29. 29.
    Emrani A-S, Danesh N-M, Ramezani M, Taghdisi S-M, Abnous K (2016) A novel fluorescent aptasensor based on hairpin structure of complementary strand of aptamer and nanoparticles as a signal amplification approach for ultrasensitive detection of cocaine. Biosens Bioelectron 79:288–293CrossRefGoogle Scholar
  30. 30.
    Roushani M, Shahdost-fard F (2015) A novel ultrasensitive aptasensor based on silver nanoparticles measured via enhanced voltammetric response of electrochemical reduction of riboflavin as redox probe for cocaine detection. Sensors Actuators B-Chem 207:764–771CrossRefGoogle Scholar
  31. 31.
    Nie J, Deng Y, Deng Q-P, Zhang D-W, Zhou Y-L, Zhang X-X (2013) A self-assemble aptamer fragment/target complex based high-throughput colorimetric aptasensor using enzyme linked aptamer assay. Talanta 106:309–314CrossRefGoogle Scholar
  32. 32.
    Zhang H-X, Jiang B-Y, Xiang Y, Zhang Y-Y, Chai Y-Q, Yuan R (2011) Aptamer/quantum dot-based simultaneous electrochemical detection of multiple small molecules. Anal Chim Acta 688:99–103CrossRefGoogle Scholar
  33. 33.
    Zhou J-W, Ellis A-V, Kobus H, Voelcker N-H (2012) Aptamer sensor for cocaine using minor groove binder based energy transfer. Anal Chim Acta 719:76–81CrossRefGoogle Scholar
  34. 34.
    Roushani M, Shahdost-Fard F (2015) A highly selective and sensitive cocaine aptasensor based on covalent attachment of the aptamer-functionalized aunps onto nanocomposite as the support platform. Anal Chim Acta 853:214–221CrossRefGoogle Scholar
  35. 35.
    Roncancio D, H-X Y, X-W X, Wu S, Liu R, Debord J, Lou X-H, Xiao Y (2014) A label-free aptamer-fluorophore assembly for rapid and specific detection of cocaine in biofluids. Anal Chem 86:11100–11106CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  1. 1.School of Laboratory MedicineHubei University of Chinese MedicineWuhanPeople’s Republic of China
  2. 2.Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and MoxibustionWuhanPeople’s Republic of China
  3. 3.Teaching and Research Office of Forensic MedicineHubei University of Chinese MedicineWuhanPeople’s Republic of China

Personalised recommendations