Analytical and Bioanalytical Chemistry

, Volume 410, Issue 16, pp 3661–3669 | Cite as

Magnetic bead/capture DNA/glucose-loaded nanoliposomes for amplifying the glucometer signal in the rapid screening of hepatitis C virus RNA

  • Haijian Tu
  • Kun Lin
  • Yongzhi Lun
  • Liuming Yu
Paper in Forefront


A digital detection strategy based on a portable personal glucometer (PGM) was developed for the simple, rapid, and sensitive detection of hepatitis C virus (HCV) RNA, involving the release of glucose-loaded nanoliposomes due to coupling-site-specific cleavage by the endonuclease BamHI. The glucose-loaded nanoliposomes were synthesized using a reversed-phase evaporation method and provided an amplified signal at the PGM in the presence of HCV RNA. Initially, a 21-mer oligonucleotide complementary to HCV RNA was covalently conjugated to a magnetic bead through the amino group at the 5′ end of the oligonucleotide, and then bound to a glucose-loaded liposome by typical carbodiimide coupling at its 3′ end. In the presence of the target HCV RNA, the target hybridized with the oligonucleotide to form double-stranded DNA. The symmetrical duplex sequence 5′-GGATCC-3′ between guanines was then catalytically cleaved by BamHI, which detached the glucose-loaded liposome from the magnetic bead. Following magnetic separation of the bead, the detached glucose-loaded liposome was lysed using Triton X-100 to release the glucose molecules within it, which were then detected as an amplified signal at the digital PGM. Under optimal conditions, the PGM signal increased with increasing HCV RNA, and displayed a strongly linear dependence on the level of HCV RNA for concentrations ranging from 10 pM to 1.0 μM. The detection limit (LOD) of the system was 1.9 pM. Good reproducibility and favorable specificity were achieved in the analysis of the target HCV RNA. Human serum samples containing HCV RNA were analyzed using this strategy, and the developed sensing platform was observed to yield satisfactory results based on a comparison with the corresponding results from a Cobas® Amplicor HCV Test Analyzer.

Graphical abstract

A digital detection strategy utilizing a personal glucometer was developed for the detection of hepatitis C virus RNA. The strategy involved the use of the endonuclease BamHI along with a 21-mer oligonucleotide conjugated to both a magnetic bead and a glucose-loaded nanoliposome. Hybridization of the nucleotide with the target RNA triggered the coupling-site-specific cleavage of the duplex by BamHI, leading to the release of the glucose-loaded nanoliposome. Following separation of the magnetic bead, the free nanoliposome was dissolved, liberating the glucose molecules within it, which in turn were detected as an amplified signal by the glucometer


Digital detection strategy Hepatitis C virus RNA Glucose-loaded liposome Personal glucometer BamHI endonuclease 



This work was financially supported by the National Natural Science Foundation of China (grant no.: 81703477), and the Earmarked Funds Provided by the Science and Technology Plan Projects of Putian City (grant no.: 2018).

Compliance with ethical standards

All procedures performed in studies involving human participants were approved by the Affiliated Hospital of Putian University and were carried out in accordance with the ethical standards of the Fujian Provincial Research Committee and the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained individually from all participants included in this study.

Conflict of interest

The authors declare that they have no competing interests.


  1. 1.
    Shahid I, AlMalki W, Hassan S, Hafeez M. Real-world challenges for hepatitis C virus medications: a critical overview. Crit Rev Microbiol. 2018;44:143–60.CrossRefPubMedGoogle Scholar
  2. 2.
    Poe A, Duong N, Bedi K, Kodani M. Stability of hepatitis C virus RNA and anti-HCV antibody in air-dried and freeze-dried human plasma samples. J Virol Methods. 2018;253:53–5.CrossRefPubMedGoogle Scholar
  3. 3.
    Li Y, Yamane D, Masaki T, Lemon S. The yin and yang of hepatitis C: synthesis and decay of hepatitis C virus RNA. Nat Rev Microbiol. 2015;13:544–58.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Campos-Varela I, Agudelo E, Sarkar M, Roberts J, Terrault N. Use of a hepatitis C virus (HCV) RNA-positive donor in a treated HCV RNA-negative liver transplant recipient. Transplant Infectious Disease. 2018;20:e12809.Google Scholar
  5. 5.
    Sarnow P, Sagan S. Unraveling the mysterious interactions between hepatitis C virus RNA and liver-specific microRNA-122. Ann Rev Virol. 2016;3:309–32.CrossRefGoogle Scholar
  6. 6.
    Shawky S, Awad A, Allam W, Alkordi M, El-Khamisy S. Gold aggregating gold: a novel nanoparticle biosensor approach for the direct quantification of hepatitis C virus RNA in clinical samples. Biosens Bioelectron. 2017;92:349–56.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Al Olaby R, Azzazy H. Hepatitis C virus RNA assays: current and emerging technologies and their clinical applications. Expert Rev Molec Diagnos. 2011;11:53–64.CrossRefGoogle Scholar
  8. 8.
    Jang J, Lee K. Rapid detection of serum HCV RNA by combining reverse transcription and PCR without RNA extraction. Arch Pharm Res. 1996;19:486–9.CrossRefGoogle Scholar
  9. 9.
    Chen J, Xu M, Huang Z, Luo Y, Tang L, Jiang J. BEAMing LAMP: single-molecule capture and on-bead isothermal amplification for digital detection of hepatitis C virus in plasma. Chem Commun. 2018;54:291–4.CrossRefGoogle Scholar
  10. 10.
    Yang Y, Wang Y, Fang Z, Yu Y, Yong Y. Bioelectrochemical biosensor for water toxicity detection: generation of dual signals for electrochemical assay confirmation. Anal Bioanal Chem. 2018;410:1231–6.CrossRefPubMedGoogle Scholar
  11. 11.
    Shu J, Tang D. Current advances in quantum-dots-based photoelectrochemical immunoassay. Chem Asian J. 2017;12:2780–9.CrossRefPubMedGoogle Scholar
  12. 12.
    Shu J, Qiu Z, Lv S, Zhang TD. Plasmonic enhancement coupling with defect-engineered TiO2−x: a mode for sensitive photoelectrochemical biosensing. Anal Chem. 2018;90:2425–9.Google Scholar
  13. 13.
    Sharma S, Singh N, Tomar V, Chandra R. A review on electrochemical detection of serotonin based on surface modified electrode. Biosens Bioelectron. 2018;107:76–93.CrossRefPubMedGoogle Scholar
  14. 14.
    Puiu M, Bala C. Petide-based biosensors: from self-assembly interfaces to molecular probes in electrochemical assays. Bioelectrochemistry. 2018;120:66–75.CrossRefPubMedGoogle Scholar
  15. 15.
    Wen W, Yan X, Zhu C, Du D, Lin Y. Recent advances in electrochemical immunosensors. Anal Chem. 2017;89:138–56.CrossRefPubMedGoogle Scholar
  16. 16.
    Gao Z, Tang D, Xu M, Chen G, Yang H. Nanoparticle-based pseudo hapten for target-responsive cargo release from a magnetic mesoporous silica nanocontainers. Chem Commun. 2014;50:6256–8.CrossRefGoogle Scholar
  17. 17.
    Poitout V, Moatti-Sirat D, Reach G. Calibration in dogs of a subcutaneous miniaturized glucose sensor using a glucose meter for blood glucose determination. Biosens Bioelectron. 1992;7:587–92.CrossRefPubMedGoogle Scholar
  18. 18.
    Deng K, Zhang Y, Tong X. Sensitive electrochemical detection platform for target microRNA-21 based on propylamine-functionalized mesoporous silica with glucometer readout. Anal Bioanal Chem. 2018;410:1863–71.CrossRefPubMedGoogle Scholar
  19. 19.
    Tang J, Huang Y, Liu H, Zhang C, Tang D. Novel glucometer-based immunosensing strategy suitable for complex systems with signal amplification using surfactant-responsive cargo release from glucose-encapsualted lipsome nanocarriers. Biosens Bioelectron. 2016;79:508–14.CrossRefPubMedGoogle Scholar
  20. 20.
    Qiu Z, Shu J, Tang D. Near-infrared-to-ultravoilet light-mediated photoelectrochemical aptasensing platform for cancer biomarker based on core-shell NaYF4:Yb,Tm@TiO2 upconversion microrods. Anal Chem. 2018;90:1021–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Zhou Q, Lin Y, Zhang K, Li M, Tang D. Reduced graphene oxide/BiFeO3 nanohybrids-based signal-on photoelectrochemical sensing system for prostate-specific antigen detection coupling with magnetic microfluidic device. Biosens Bioelectron. 2018;101:146–52.CrossRefPubMedGoogle Scholar
  22. 22.
    Lin Y, Zhou Q, Tang D, Niessner R, Kopp D. Signal-on photoelectrochemical immunoassay for aflatoxin B1 based on enzymatic product-etching MnO2 nanosheets for dissociation of carbon dots. Anal Chem. 2017;89:5637–45.CrossRefPubMedGoogle Scholar
  23. 23.
    Reverte L, Prieto-Simon B, Campas M. New advances in electrochemical biosensors for the detection of toxins: nanomaterials, magnetic beads and microfluidics system. A review. Anal Chim Acta. 2016;908:8–21.Google Scholar
  24. 24.
    Tekin H, Gijs M. Ultrasensitive protein detection: a case for microfluidic magnetic bead-based assays. Lab Chip. 2013;13:4711–39.CrossRefPubMedGoogle Scholar
  25. 25.
    Scherr T, Markwalter C, Bauer W, Gasperino D, Wright D, Haselton F. Application of mass transfer theory to biomarker capture by surface functionalized magnetic beads in microcentrifuge tubes. Adv Colloid Interf Sci. 2017;246:275–88.CrossRefGoogle Scholar
  26. 26.
    Lin Y, Zhou Q, Tang D. Dopamine-loaded liposomes for in-situ amplified photoelectrochemical immunoassay of AFB1 to enhance photocurrent of Mn2+-doped Zn3(OH)2V2O7 nanobelts. Anal Chem. 2017;89:11803–10.CrossRefPubMedGoogle Scholar
  27. 27.
    Tang D, Su B, Tang J, Ren J, Chen G. Nanoparticle-based sandwich electrochemical immunoassay for carbohydrate antigen 125 with signal enhancement using enzyme-coated nanometer-sized enzyme-doped silica beads. Anal Chem. 2010;82:1527–34.CrossRefPubMedGoogle Scholar
  28. 28.
    Israelachvili J, Mitechell D. A model for the packing of lipids in bilayer membranes. Biochim Biophys Acta. 1975;389:13–29.CrossRefPubMedGoogle Scholar
  29. 29.
    Griffin J, Singh A, Senapati D, Rhodes P, Mitchell K, Robinson B, et al. Size- and distance-dependent nanoparticle surface-energy transfer (NSET) method for selective sensing of hepatitis C virus RNA. Chem Eur J. 2009;15:342–51.CrossRefPubMedGoogle Scholar
  30. 30.
    Su H, Meng X, Guo Q, Tan Y, Cai Q, Qin H, et al. Label-free DNAsensor with PCR-like sensitivity based on background reduction and target-triggered polymerization amplification. Biosens Bioelectron. 2014;52:417–21.CrossRefPubMedGoogle Scholar
  31. 31.
    Lu M, Xu L, Zhang X, Xiao R, Wang Y. Ag(I)-coordinated hairpin DNA for homogenous electronic monitoring of hepatitis C virus accompanying isothermal cycling signal amplification strategy. Biosens Bioelectron. 2015;73:195–201.CrossRefPubMedGoogle Scholar
  32. 32.
    Tang D, Tang J, Su B, Li Q, Chen G. Electrochemical detection of hepatitis C virus with signal amplification using BamHI endonuclease and horseradish peroxidase-encapsulated nanogold hollow spheres. Chem Commun. 2011;47:9477–9.CrossRefGoogle Scholar
  33. 33.
    Guillou-Guillemette L, Lunel-Fabiani F. Detection and quantification of serum or plasma HCR RNA: mini review of commercially available assays. In: Tang H, editor. Heptatitis C: methods and protocols, vol. 510. 2nd ed. Totowa, NJ: Humana Press; 2009. p. 3–14.Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.The Affiliated Hospital of Putian UniversityPutian UniveristyPutianChina
  2. 2.School of Pharmacy and Medical TechnologyPutian UniversityPutianChina

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