In this research, we report highly sensitive and specific sandwich-type SPR-based biosensor for the detection H5Nx whole viruses. A few of aptamers, for the first time, were successfully screened and characterized for whole avian influenza (AI) viruses, H5Nx, by using Multi-GO-SELEX method. The affinities of the aptamers developed in this study were ranged from 8×104 to 1×104 EID50/ml, and the aptamers IF22, IF23 were found to be specific to H5N1 and H5N8, respectively. In addition, some flexible aptamers IF20, IF15, and IF10 were found to bind to the H5N1 and H5N2, H5N1 and H5N8, or H5N1, H5N2, and H5N8, respectively. Moreover, aptamers IF10 and IF22 were found to bind H5N1 virus simultaneously and confirmed to bind the different site of the same H5N1 whole virus. Therefore, this pair of aptamers, IF10 and IF22, were successfully applied to develop the sandwich-type SPR-based biosensor assay which is rapid, accurate for the detection of AI whole virus from H5N1-infected feces samples. The minimum detectible concentration of H5N1 whole virus was found to be 200 EID50/ml with this sandwich-type detection using the aptamer pair obtained in this study. In addition, the sensitivity of this biosensor was successfully enhanced by using the signal amplification with the secondary aptamer conjugated with gold nanoparticles.
Bibliographical noteFunding Information:
This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea Government (MSIP) (No. 2016R1E1A2020541 and No. 2016R1A2B3011422 ), the Basic Core Technology Development Program for the Oceans and the Polar Regions of the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning ( 2015M1A5A1037055 ) and the KIST Institutional Program (Project No. 2E00000-00-000 ). The Dr. V. T Nguyen's salary was financially supported by Korea University Grant.
© 2016 Elsevier B.V.
- Circular Dichroism
- Dual aptamer
- Whole virus H5Nx
ASJC Scopus subject areas
- Biomedical Engineering