Biosensors and Bioelectronics. 2015 Feb;64:311-317

A Plasmon-Assisted Fluoro-Immunoassay using Gold Nanoparticle-Decorated Carbon Nanotubes for Monitoring the Influenza Virus.

Jaewook Lee1,5, Syed Rahin Ahmed2,3, Sangjin Oh3, Jeonghyo Kim3, Tetsuro Suzuki4, Kaushik Parmar5, Simon S. Park5, Jaebeom Lee3,*, and Enoch Y. Park1,2**

1Research Institute of Green Science and Technology, Shizuoka University 836 Ohya Suruga-ku, Shizuoka 422-8529, Japan

2Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, 836 Ohya Suruga- ku, Shizuoka 422-8529, Japan

3Department of Nano Fusion and Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Korea

4Department of Infectious Diseases, Hamamatsu University School of Medicine, 1-20-1 Higashi-ku, Handa-yama, Hamamatsu 431-3192, Japan

5Department of Mechanical and Manufacturing Engineering, University of Calgary, Calgary, T2N 1N4, Canada

 

ABSTRACT

A plasmon-assisted fluoro-immunoassay (PAFI) was developed for the detection of the influenza virus by using Au nanoparticle (Au NP)-decorated carbon nanotubes (AuCNTs) that were synthesized using phytochemical composites at room temperature in deionized water. Specific antibodies (Abs) against the influenza virus were conjugated onto the surface of AuCNTs and cadmium telluride quantum dots (QDs), which had a photoluminescence intensity that varied as a function of virus concentration and a detection limit of 0.1 pg/mL for all three types of influenza viruses examined. The clinically isolated influenza viruses (A/Yokohama/110/2009 (H3N2)) were detected in the range of 50–10,000 PFU/mL, with a detection limit of 50 PFU/mL. From a series of proof-of-concept and clinical experiments, the developed PAFI biosensing system provided robust signal production and enhancement, as well as an excellent selectivity and sensitivity for influenza viruses. This nanoparticle-based technique could be potentially developed as an efficient detection platform for the influenza virus.

KEYWORDS: Plasmon-assisted fluoro-immunoassay, Gold nanoparticle-decorated carbon nanotube, CdTe quantum dot, Influenza virus detection platform, Plasmonic resonance energy transfer

PMID: 25240957

 

Supplement:

Recently, fluoro-immunoassay methodologies have been applied for biomolecule monitoring, such as virus diagnosis, DNA detection, enzyme monitoring and so on.1 Especially, plasmonic assisted fluoro-immunoassay (PAFI) based detection strategy has been spotlighted due to their high selectivity and sensitivity.2 In order to develop and demonstrate such PAFI, metal nanoparticles (NPs) modified carbon nanostructures (CNS) and fluorescent NPs were used as nanobiosensing platforms. In this case, metal NPs decorated CNS plays a role as plasmonic generated materials and fluorescent NPs performs as enhanced fluorescence display source for visualized monitoring.

This detection mechanism is illustrated in Figure 1 (A). In our case, antibody conjugated two nanomaterials were used for influenza virus detection; Au-CNTs were used as plasmonic materials and QDs were applied as fluorescent NPs. If target influenza virus were existed in Au-CNTs and QDs system, both nanomaterials could be hybridized with biomolecules due to their antigen-antibody reaction. And as a result, fluorescence of sandwich type-assembly structure was observed in Au-CNTs and QDs hybrid system (Figure (B) – (D)).

 JL fig1

Figure 1. (A) Schematic illustration of detection process using Au-CNT and QDs (non-scalable), fluorescent image of virus induced Au-CNT/QDs hybrid structure; (B) dark field, (C) bright field and (D) merged image via confocal microscope.

 

In addition, fluorescence intensity variation was occurred depending on the virus concentrations. In the other words, as increasing the virus concentration, fluorescence intensity was also increased. And, based on these variation results, quantity of virus could be analyzed. In this study, we demonstrated three different kinds of influenza virus (influenza virus A/Beijing/262/95 (H1N1), influenza virus/New Caledonia/20/99IvR116 (H1N1) and clinically isolated sample (influenza virusA/Yokohama/110/2009(H3N2))) were monitored and as a result, H1N1 virus detection limitation of this system was 0.1 pg/mL. Furthermore, influenza virus in clinical sample was detected using PAFI with 50 PFU/mL detection limitation. Indeed, selectivity property could be tuned by antibody change on the nanomaterials surface. By using H1N1 influenza virus targeted antibody conjugated nanomaterials, only clinically isolated samples were monitored, but H3N2 virus was not detected (Figure 2).

JL fig2Figure 2. Selectivity monitoring with H1N1-targeted antibody conjugated AuCNTs and QDs

 

Thus, potential diagnostic system of this PAFI system were recognized with influenza virus monitoring and these nanomaterials based sensing platform could be applied not only influenza virus detection but also other biomolecules monitoring.

 

References

  1. Fengming Zou, Hongjian Zhou, Van Tan Tran, Jeonghyo Kim, Kwangnak Koh, Jaebeom Lee., ACS Appl. Mater. Interfaces, 2015;7:12168–12175
  2. Jaewook Lee, Jeonghyo Kim, Syed Rahin Ahmed, Hongjian Zhou, Jong-Man Kim, Jaebeom Lee., ACS Appl. Mater. Interfaces, 2014;6:21380-21388

 

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