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. 2012;12(9):12506-18.
doi: 10.3390/s120912506. Epub 2012 Sep 13.

A SPR aptasensor for detection of avian influenza virus H5N1

Hua Bai  1 , Ronghui Wang, Billy Hargis, Huaguang Lu, Yanbin Li
Affiliations

A SPR aptasensor for detection of avian influenza virus H5N1

Hua Bai et al. Sensors (Basel). 2012.

Abstract

Rapid and specific detection of avian influenza virus (AIV) is urgently needed due to the concerns over the potential outbreaks of highly pathogenic H5N1 influenza in animals and humans. Aptamers are artificial oligonucleic acids that can bind specific target molecules, and show comparable affinity for target viruses and better thermal stability than monoclonal antibodies. The objective of this research was to use a DNA-aptamer as the specific recognition element in a portable Surface Plasmon Resonance (SPR) biosensor for rapid detection of AIV H5N1 in poultry swab samples. A SPR biosensor was fabricated using selected aptamers that were biotinylated and then immobilized on the sensor gold surface coated with streptavidin via streptavidin-biotin binding. The immobilized aptamers captured AIV H5N1 in a sample solution, which caused an increase in the refraction index (RI). After optimizing the streptavidin and aptamer parameters, the results showed that the RI value was linearly related (R(2) = 0.99) to the concentration of AIV in the range of 0.128 to 1.28 HAU. Negligible signal (<4% of H5N1) was observed from six non-target AIV subtypes. The AIV H5N1 in poultry swab samples with concentrations of 0.128 to 12.8 HAU could be detected using this aptasensor in 1.5 h.

Keywords: aptamer; avian influenza virus; biosensor; surface plasmon resonance.

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Figures

Figure 1.
Figure 1.
Sequence and predicted secondary structure of the aptamer used in this study.
Figure 2.
Figure 2.
The configuration for measuring AIV using the portable SPR aptasensor.
Figure 3.
Figure 3.
Principle of SPR biosensor for detection of AIV H5N1: (a) Streptavidin immobilization; (b) Biotinylated aptamer immobilization; (c) Virus detection.
Figure 4.
Figure 4.
Optimization of streptavidin and aptamers, each concentration and incubation time repeated three times, respectively: (a) Streptavidin concentration; (b) Streptavidin incubation time; (c) Aptamer concentration; (d) Aptamer incubation time.
Figure 5.
Figure 5.
Detection of AIV H5N1 using SPR aptasensor: (a) A typical response curve of the SPR aptasensor to the surface modification and AIV H5N1 detection; (b) Calibration curve for AIV H5N1 detection (all tests were repeated three times).
Figure 6.
Figure 6.
SPR aptasensor for detection of AIV H5N1 in poultry swab samples.
See this image and copyright information in PMC

References

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