Development of Immunoassays for Detection of Francisella tularensis Lipopolysaccharide in Tularemia Patient Samples
| Autor: | Yasemin Ozsurekci, Jose Arias-Umana, Marcellene A. Gates-Hollingsworth, Heather R. Green, Peter Thorkildson, David P. AuCoin, Haley L. DeMers, Teerapat Nualnoi, Anjana Dissanayaka, Kayleigh J. Robichaux, Derrick Hau, Kathryn J. Pflughoeft, Emily E. Hannah, Sujata G. Pandit |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Zoonotic Bacterial Infection Microbiological culture LPS 030106 microbiology Population diagnostic patient samples Article Serology Tularemia 03 medical and health sciences Immunology and Allergy Medicine antibodies Francisella tularensis education Molecular Biology education.field_of_study General Immunology and Microbiology biology business.industry Infectious dose lipopolysaccharide lateral flow immunoassay Outbreak biology.organism_classification medicine.disease bacterial infections and mycoses Virology tularemia 030104 developmental biology Infectious Diseases monoclonal antibodies enzyme-linked immunosorbent assay business |
| Zdroj: | Pathogens Volume 10 Issue 8 Pathogens, Vol 10, Iss 924, p 924 (2021) |
| ISSN: | 2076-0817 |
| DOI: | 10.3390/pathogens10080924 |
| Popis: | Francisella tularensis is the causative agent of tularemia, a zoonotic bacterial infection that is often fatal if not diagnosed and treated promptly. Natural infection in humans is relatively rare, yet persistence in animal reservoirs, arthropod vectors, and water sources combined with a low level of clinical recognition make tularemia a serious potential threat to public health in endemic areas. F. tularensis has also garnered attention as a potential bioterror threat, as widespread dissemination could have devastating consequences on a population. A low infectious dose combined with a wide range of symptoms and a short incubation period makes timely diagnosis of tularemia difficult. Current diagnostic techniques include bacterial culture of patient samples, PCR and serological assays however, these techniques are time consuming and require technical expertise that may not be available at the point of care. In the event of an outbreak or exposure a more efficient diagnostic platform is needed. The lipopolysaccharide (LPS) component of the bacterial outer leaflet has been identified previously by our group as a potential diagnostic target. For this study, a library of ten monoclonal antibodies specific to F. tularensis LPS were produced and confirmed to be reactive with LPS from type A and type B strains. Antibody pairs were tested in an antigen-capture enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay format to select the most sensitive pairings. The antigen-capture ELISA was then used to detect and quantify LPS in serum samples from tularemia patients for the first time to determine the viability of this molecule as a diagnostic target. In parallel, prototype lateral flow immunoassays were developed, and reactivity was assessed, demonstrating the potential utility of this assay as a rapid point-of-care test for diagnosis of tularemia. |
| Databáze: | OpenAIRE |
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