title: Highly sensitive detection of a bio-threat pathogen by gold nanoparticle-based oligonucleotide-linked immunosorbent assay.
abstract: Francisella (F.) tularensis causes the zoonotic disease tularemia and categorized as one of the highest-priority biological agents. The sensing approaches utilized by conventional detection methods, including enzyme-linked immunosorbent assay (ELISA), are not sensitive enough to identify an infectious dose of this high-risk pathogen due to its low infective dose. As an attempt to detect F. tularensis with high sensitivity, we utilized the highly sensitive immunoassay system named gold nanoparticle-based oligonucleotide-linked immunosorbent assay (GNP-OLISA) which uses antibody-gold nanoparticles conjugated with DNA strands as a signal generator and RNA oligonucleotides appended with a fluorophore as a quencher for signal amplification. We modified the GNP-OLISA for the detection F. tularensis to utilize one antibody for both the capture of the target and for signal generation instead of using two different antibodies, which are usually employed to construct the antibody sandwich in the ELISA. The GNP-OLISA showed 37-fold higher sensitivity compared with ELISA and generated very consistent detection results in the sera. In addition, the detection specificity was not affected by the presence of non-target bacteria, suggesting that GNP-OLISA can be used as a sensitive detection platform for monitoring high-risk pathogens thereby overcoming the limit of the conventional assay system.
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Seo
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Young-Ran
authors.initials: YR
authors.lastname: Lee
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Jun
authors.initials: J
authors.lastname: Ho Jeon
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Yi-Rang
authors.initials: YR
authors.lastname: Hwang
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Pil-Gu
authors.initials: PG
authors.lastname: Park
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, KIST Campus, Korea University of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Dae-Ro
authors.initials: DR
authors.lastname: Ahn
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Ki-Cheol
authors.initials: KC
authors.lastname: Han
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea.
authors.forename: Gi-Eun
authors.initials: GE
authors.lastname: Rhie
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheongwon, Chungcheongbuk-do 363-951, Republic of Korea. Electronic address: khong@nih.go.kr.
authors.forename: Kee-Jong
authors.initials: KJ
authors.lastname: Hong
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005604:Francisella tularensis / Q000302:isolation & purification*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D061925:Immobilized Nucleic Acids / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D007164:Immunosorbents / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D014406:Tularemia / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D055910:Antibodies, Immobilized; D061925:Immobilized Nucleic Acids; D007164:Immunosorbents; D006046:Gold
keywords: Bacteria; Detection sensitivity; ELISA; Francisella tularensis; Gold-nanoparticle; OLISA