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Bluetongue Virus Detection Using Microspheres Conjugated with Monoclonal Antibodies against Group-Specific Protein Vp7 by Flow Virometry

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Abstract—

Bluetongue is a reemerging transmissive infectious disease that threatens all countries with intensive livestock production. In cattle, bluetongue may either lead to lethality or be asymptomatic with a long-lasting viremic phase, which makes development of high-sensitivity bluetongue virus detection methods an important task. Methods of virus particle detection based on immunofluorescence assay in bluetongue-infected Vero cells, sandwich enzyme immunoassay and flow virometry have been developed using high-affinity monoclonal antibodies against VP7 (viral protein 7). High analytical sensitivity of the method (10–0.25 TCID50/mL (tissue cytopathogenic dose)) was demonstrated in a flow cytometry assay. The ability to rapidly and efficiently detect virus particles in the biological fluids obtained from animals by flow virometry was demonstrated in a model system in which BTV was measured in cattle blood serum. The detection sensitivity was 100.75 TCID50/mL.

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Funding

The work was supported by the Russian Academy of Sciences State Task AAAA-A19-119050790041-1, topic ID 0101-2019-0038.

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Authors and Affiliations

  1. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Pushchino Branch, 142290, Pushchino, Russia

    N. V. Rudenko, A. P. Karatovskaya, A. V. Zamyatina, F. A. Brovko & A. O. Shepelyakovskaya

  2. Federal Research Center for Virology and Microbiology, 601125, Volginsky Settlement, Vladimir oblast, Russia

    A. S. Malogolovkin, A. U. Kol’tsov, O. G. Lapteva & D. V. Kolbasov

  3. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

    V. A. Oleinikov

  4. Sirius University of Science and Technology, 354340, Sirius Settlement, Sirius federal territory, Krasnodar krai, Russia

    A. S. Malogolovkin

Authors
  1. N. V. Rudenko
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  2. A. P. Karatovskaya
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  3. A. V. Zamyatina
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  4. A. S. Malogolovkin
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  5. V. A. Oleinikov
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  6. F. A. Brovko
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  7. A. U. Kol’tsov
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  8. O. G. Lapteva
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  9. D. V. Kolbasov
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  10. A. O. Shepelyakovskaya
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Correspondence to N. V. Rudenko.

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COMPLIANCE WITH ETHICAL STANDARDS

No experimentation involving animals or human was performed by any of the authors.

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Additional information

Translated by E. Martynova

Abbreviations: BTV, bluetongue virus; FBS, fetal bovine serum; FITC, fluorescein isothiocyanate; mAb, monoclonal antibodies; MIF, median intensity of fluorescence; MOI, multiplicity of infection; PE, phycoerythrin; SA-PE, phycoerythrin-labeled streptavidin; Sulfo-SMCC, sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate; VP7, virus protein 7; IIFA, indirect immunofluorescence assay; TCID, tissue culture infective dose.

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Rudenko, N.V., Karatovskaya, A.P., Zamyatina, A.V. et al. Bluetongue Virus Detection Using Microspheres Conjugated with Monoclonal Antibodies against Group-Specific Protein Vp7 by Flow Virometry. Russ J Bioorg Chem 48, 793–800 (2022). https://doi.org/10.1134/S1068162022040173

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