Bioluminescent detection probe for tick-borne encephalitis virus immunoassay

  • Burakova L
  • Kudryavtsev A
  • Stepanyuk G
 et al. 
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Abstract

To facilitate the detection of the tick-borne encephalitis virus (TBEV), the causative agent of one of the most severe human neuroinfections, we have developed an immunoassay based on bioluminescent hybrid protein 14D5a-Rm7 as a detection probe. The protein containing Renilla luciferase as a reporter and a single-chain variable fragment (scFv) of murine immunoglobulin to TBEV as a recognition element was constructed, produced by bacterial expression, purified, and tested. Both domains were shown to reveal their specific biological properties-affinity to the target antigen and bioluminescent activity. Hybrid protein was applied as a label for solid-phase immunoassay of the antigens, associated with the tick-borne encephalitis virus (native glycoprotein E or extracts of the infected strain of lab ticks). The assay demonstrates high sensitivity (0.056 ng of glycoprotein E; 10(4)-10(5) virus particles or 0.1 pg virions) and simplicity and is competitive with conventional methods for detection of TBEV.

Author-supplied keywords

  • Immunoassay
  • Luciferase
  • Single-chain antibody
  • Tick-borne encephalitis virus

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Authors

  • Ludmila P. Burakova

  • Alexander N. Kudryavtsev

  • Galina A. Stepanyuk

  • Ivan K. Baykov

  • Vera V. Morozova

  • Nina V. Tikunova

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