title: Highly sensitive sandwich-type SPR based detection of whole H5Nx viruses using a pair of aptamers.
abstract: 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 8×10(4) to 1×10(4)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.
authors.affiliation: Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-713, Republic of Korea.
authors.forename: Van-Thuan
authors.initials: VT
authors.lastname: Nguyen
authors.affiliation: Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-713, Republic of Korea.
authors.forename: Ho Bin
authors.initials: HB
authors.lastname: Seo
authors.affiliation: Center for Environment, Health and Welfare Research, Korea Institute of Science and Technology (KIST), Department of Energy and Environmental Engineering, Korea University of Science and Technology (UST), Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Byoung Chan
authors.initials: BC
authors.lastname: Kim
authors.affiliation: Center of BioMicrosystems,Korea Institute of Science and Technology (KIST), Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Sang Kyung
authors.initials: SK
authors.lastname: Kim
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, Konkuk University, Kwangjin-gu, Seoul 143-701, Republic of Korea.
authors.forename: Chang-Seon
authors.initials: CS
authors.lastname: Song
authors.affiliation: Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Anam-dong, Seongbuk-gu, Seoul 136-713, Republic of Korea. Electronic address: mbgu@korea.ac.kr.
authors.forename: Man Bock
authors.initials: MB
authors.lastname: Gu
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D052157:Aptamers, Nucleotide; D004279:DNA, Viral
keywords: AuNP; Circular Dichroism; Dual aptamer; SPR; Whole virus H5Nx