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Development of an immunoassay test system based on monoclonal antybodies and immunomagnetic particles for the detection of F. tularensis cells.
Vetchinin, S S; Shevyakov, Anton Georgievich; Khomyakov, A E; Mironova, R I; Mokrievich, A N; Biketov, S F.
Afiliação
  • Vetchinin SS; State Research Center of Applied Microbiology and Biotechnology.
  • Shevyakov AG; State Research Center of Applied Microbiology and Biotechnology.
  • Khomyakov AE; State Research Center of Applied Microbiology and Biotechnology.
  • Mironova RI; State Research Center of Applied Microbiology and Biotechnology.
  • Mokrievich AN; State Research Center of Applied Microbiology and Biotechnology.
  • Biketov SF; State Research Center of Applied Microbiology and Biotechnology.
Klin Lab Diagn ; 66(6): 353-357, 2021 Jun 07.
Article em En | MEDLINE | ID: mdl-34105911
ABSTRACT
Tularemia is an especially dangerous infection caused by the gram-negative bacterium Francisella tularensis. It belongs to natural focal infections, and therefore is under continuous control by quarantine services. When carrying out their activities they use a whole range of diagnostic tools. The objective of this research is to develop an enzyme immunoassay based on highly specific monoclonal antibodies and immunomagnetic particles for monitoring the tularemia pathogen. To produce hybridomas mice were immunized with cells of the vaccine strain F. tularensis subsp. holarctica 15 NIIEG. After cell fusion hybridomas were selected by a solid-phase enzyme immunoassay (ELISA) using lipopolysaccharide (LPS) of the tularemia microbe. As a result, two hybridomas, 1C2 and 3F5, were produced. MABs of the hybridomas were obtained by using BALB / c mice. The MABs were purified by sepharose A affinity chromatography and used for conjugation with magnetic particles, and for biotinylation followed by matching a pair for ELISA. The pair of IMPs and MABs 3F5 as well as biotinylated FB11-x MABs was the best in detecting tularemia cells. The use of this MAB pair in ELISA allowed the identification of 105 microbial cells/ml in a 4 ml sample and 5×103 microbial cells/ml in a 45ml sample. Interaction with F. tularensis subsp. novicida Utah112 cells was absent.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tularemia / Francisella tularensis Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tularemia / Francisella tularensis Idioma: En Ano de publicação: 2021 Tipo de documento: Article