A sensitive, specific and rapid method for the detection of three different kinds of plant viruses including tomato ringspot virus (ToRSV), bean pod mottle virus (BPMV) and arabis mosaic virus (ArMV) was demonstrated using novel upconversion nanoparticles (UCNPs) as a fluorescence marker coupled with immunomagnetic separation. Magnetic nanoparticles (MNPs, ~100？nm) were coated with different antibodies were employed to capture and enrich the target viruses. Then antibody-conjugated UCNPs as signal probes were added to form sandwich complexes. This was followed by a fluorescence measurement using a 980？nm laser. This method not only avoids the difficulty in simultaneous detection of multiple independent organic fluorphores that require distinct excitation wavelengths, and auto-fluorescence of biological samples due to the higher-energy excitation of QDs but also amplifies detection signal by UCNPs-tags together with easy separation of samples by magnetic forces, demonstrating the potential to be used for detecting virus in the field of environmental safety and other fields. 1. Introduction There is increasing interest in the development of nanotechnology for bioassay [1, 2]. In the past two decades, optical and magnetic materials have attracted much attention due to their importance in the fields of chemistry, biology, medical sciences, and biotechnology [3]. Fe3O4 nanoparticles are one of the most intensively studied magnetic nanoparticles and can be applied in a variety of areas, ranging from drug delivery [4] and biosensing [5], dynamic sealing [6], and cell labeling [7] to magnetic resonance imaging [8]. Moreover, MNPs are well suited for target capturing, enrichment, and isolation [9]. Accordingly, they can be used for isolating cells [10] and bacteria [11] and for removing environmental toxins such as heavy metals and chemical waste [12]. Research on fluorescent nanomaterials has also gained extensive attention in the past decade. Recently, colloidal semiconductor nanocrystals (quantum dots, QDs) have attracted many researchers due to their broad excitation, size-dependent photoluminescence with narrow emission bandwidth covering a wide spectral range, unusual photochemical stability, and relatively high photoluminescence quantum yield [13]. However, QDs excited by blue or short-wavelength UV radiation can induce autofluorescence of most biological tissues [14], which greatly decreases the sensitivity of detection. Moreover, the risk of systemic toxicity remains high, given their incorporation of heavy metals (e.g., Pb, Cd), precluding their
References
[1]
K. K. Jain, “Nanotechnology in clinical laboratory diagnostics,” Clinica Chimica Acta, vol. 358, no. 1-2, pp. 37–54, 2005.
[2]
T. Osaka, T. Matsunaga, T. Nakanishi, A. Arakaki, D. Niwa, and H. Iida, “Synthesis of magnetic nanoparticles and their application to bioassays,” Analytical and Bioanalytical Chemistry, vol. 384, no. 3, pp. 593–600, 2006.
[3]
G. Wang, Q. Peng, and Y. Li, “Lanthanide-doped nanocrystals: synthesis, optical-magnetic properties, and applications,” Accounts of Chemical Research, vol. 44, no. 5, pp. 322–332, 2011.
[4]
O. Veiseh, J. W. Gunn, and M. Zhang, “Design and fabrication of magnetic nanoparticles for targeted drug delivery and imaging,” Advanced Drug Delivery Reviews, vol. 62, no. 3, pp. 284–304, 2010.
[5]
L. Yang, X. Ren, F. Tang, and L. Zhang, “A practical glucose biosensor based on Fe3O4 nanoparticles and chitosan/nafion composite film,” Biosensors and Bioelectronics, vol. 25, no. 4, pp. 889–895, 2009.
[6]
P. Ripka, “New directions in fluxgate sensors,” Journal of Magnetism and Magnetic Materials, vol. 215, pp. 735–739, 2000.
[7]
C. Wilhelm and F. Gazeau, “Universal cell labelling with anionic magnetic nanoparticles,” Biomaterials, vol. 29, no. 22, pp. 3161–3174, 2008.
[8]
T. K. Jain, S. P. Foy, B. Erokwu, S. Dimitrijevic, C. A. Flask, and V. Labhasetwar, “Magnetic resonance imaging of multifunctional pluronic stabilized iron-oxide nanoparticles in tumor-bearing mice,” Biomaterials, vol. 30, no. 35, pp. 6748–6756, 2009.
[9]
A. Agrawal, T. Sathe, and S. Nie, “Single-bead immunoassays using magnetic microparticles and spectral-shifting quantum dots,” Journal of Agricultural and Food Chemistry, vol. 55, no. 10, pp. 3778–3782, 2007.
[10]
X. Liang, K. Xu, J. Xu, W. Chen, H. Shen, and J. Liu, “Preparation of immunomagnetic nanoparticles and their application in the separation of mouse CD34+ hematopoietic stem cells,” Journal of Magnetism and Magnetic Materials, vol. 321, no. 12, pp. 1885–1888, 2009.
[11]
Z. Shan, Q. Wu, X. Wang et al., “Bacteria capture, lysate clearance, and plasmid DNA extraction using pH-sensitive multifunctional magnetic nanoparticles,” Analytical Biochemistry, vol. 398, no. 1, pp. 120–122, 2010.
[12]
Y. F. Shen, J. Tang, Z. H. Nie, Y. D. Wang, Y. Ren, and L. Zuo, “Preparation and application of magnetic Fe3O4 nanoparticles for wastewater purification,” Separation and Purification Technology, vol. 68, no. 3, pp. 312–319, 2009.
[13]
I. L. Medintz, H. T. Uyeda, E. R. Goldman, and H. Mattoussi, “Quantum dot bioconjugates for imaging, labelling and sensing,” Nature Materials, vol. 4, no. 6, pp. 435–446, 2005.
[14]
O. Ehlert, R. Thomann, M. Darbandi, and T. Nann, “A four-color colloidal multiplexing nanoparticle system,” ACS Nano, vol. 2, no. 1, pp. 120–124, 2008.
[15]
X. Song, F. Li, J. Ma, N. Jia, J. Xu, and H. Shen, “Synthesis of fluorescent silica nanoparticles and their applications as fluorescence probes,” Journal of Fluorescence, vol. 21, no. 3, pp. 1205–1212, 2011.
[16]
M. Wang, C. Mi, Y. Zhang et al., “NIR-responsive silica-coated NaYbF4:Er/Tm/Ho upconversion fluorescent nanoparticles with tunable emission colors and their applications in immunolabeling and fluorescent imaging of cancer cells,” Journal of Physical Chemistry C, vol. 113, no. 44, pp. 19021–19027, 2009.
[17]
S. Gai, P. Yang, C. Li et al., “Synthesis of magnetic, up-conversion luminescent, and mesoporous core-shell-structured nanocomposites as drug carriers,” Advanced Functional Materials, vol. 20, no. 7, pp. 1166–1172, 2010.
[18]
M. Yu, F. Li, Z. Chen et al., “Laser scanning up-conversion luminescence microscopy for imaging cells labeled with rare-earth nanophosphors,” Analytical Chemistry, vol. 81, no. 3, pp. 930–935, 2009.
[19]
J. Chen and J. X. Zhao, “Upconversion nanomaterials: synthesis, mechanism, and applications in sensing,” Sensors, vol. 12, no. 3, pp. 2414–2435, 2012.
[20]
J. C. Boyer, L. A. Cuccia, and J. A. Capobianco, “Synthesis of colloidal upconverting NaYF4: Er3+/Yb3+ and Tm3+/Yb3+ monodisperse nanocrystals,” Nano Letters, vol. 7, no. 3, pp. 847–852, 2007.
[21]
C. Li, Z. Quan, J. Yang, P. Yang, and J. Lin, “Highly uniform and monodisperse β-NaYF4:Ln3+ (Ln = Eu, Tb, Yb/Er, and Yb/Tm) hexagonal microprism crystals: hydrothermal synthesis and luminescent properties,” Inorganic Chemistry, vol. 46, no. 16, pp. 6329–6337, 2007.
[22]
X. Liu, J. Zhao, Y. Sun et al., “Ionothermal synthesis of hexagonal-phase NaYF4:Yb3+,Er3+/Tm3+ upconversion nanophosphors,” Chemical Communications, no. 43, pp. 6628–6630, 2009.
[23]
Z. Du, B. Jin, W. Liu, L. Chen, and J. Chen, “Highly sensitive fluorescent-labeled probes and glass slide hybridization for the detection of plant RNA viruses and a viroid,” Acta Biochimica et Biophysica Sinica, vol. 39, no. 5, pp. 326–334, 2007.
[24]
Y. B. Tang, D. Xing, D. B. Zhu, and J. F. Liu, “An improved electrochemiluminescence polymerase chain reaction method for highly sensitive detection of plant viruses,” Analytica Chimica Acta, vol. 582, no. 2, pp. 275–280, 2007.
[25]
L. M. Wright, J. T. Kreikemeier, and C. J. Fimmel, “A concise review of serum markers for hepatocellular cancer,” Cancer Detection and Prevention, vol. 31, no. 1, pp. 35–44, 2007.
[26]
Y. J. Liu, D. J. Yao, H. Y. Chang, C. M. Liu, and C. Chen, “Magnetic bead-based DNA detection with multi-layers quantum dots labeling for rapid detection of Escherichia coli O157:H7,” Biosensors and Bioelectronics, vol. 24, no. 4, pp. 558–565, 2008.
[27]
M. Nichkova, D. Dosev, S. J. Gee, B. D. Hammock, and I. M. Kennedy, “Multiplexed immunoassays for proteins using magnetic luminescent nanoparticles for internal calibration,” Analytical Biochemistry, vol. 369, no. 1, pp. 34–40, 2007.
[28]
Y. Zhao, C. Hao, W. Ma et al., “Magnetic bead-based multiplex DNA sequence detection of genetically modified organisms using quantum dot-encoded silicon dioxide nanoparticles,” Journal of Physical Chemistry C, vol. 115, no. 41, pp. 20134–20140, 2011.
[29]
H. Shen, Y. Wang, H. Yang, and J. Jiang, “Covalent immobilization of oligoDNA on the surface of magnetic nanoparticles and surface-enhanced Raman scattering study,” Chinese Science Bulletin, vol. 48, no. 24, pp. 2698–2702, 2003.
[30]
H. H. Yang, S. Q. Zhang, X. L. Chen, Z. X. Zhuang, J. G. Xu, and X. R. Wang, “Magnetite-containing spherical silica nanoparticles for biocatalysis and bioseparations,” Analytical Chemistry, vol. 76, no. 5, pp. 1316–1321, 2004.
[31]
X. Liu, J. Xing, Y. Guan, G. Shan, and H. Liu, “Synthesis of amino-silane modified superparamagnetic silica supports and their use for protein immobilization,” Colloids and Surfaces A, vol. 238, no. 1–3, pp. 127–131, 2004.
[32]
J. Zhang and R. D. K. Misra, “Magnetic drug-targeting carrier encapsulated with thermosensitive smart polymer: core-shell nanoparticle carrier and drug release response,” Acta Biomaterialia, vol. 3, no. 6, pp. 838–850, 2007.
[33]
J. Ma, Q. Fan, L. Wang, N. Jia, Z. Gu, and H. Shen, “Synthesis of magnetic and fluorescent bifunctional nanocomposites and their applications in detection of lung cancer cells in humans,” Talanta, vol. 81, no. 4-5, pp. 1162–1169, 2010.
