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Molecular Biology

, Volume 52, Issue 4, pp 629–635 | Cite as

Immuno-PCR Assay for Quantitation of Antibodies to Epstein–Barr Virus

  • V. D. Pivovarov
  • D. Yu. Ryazantsev
  • M. A. Simonova
  • T. V. Yegorova
  • S. V. Khlgatian
  • S. K. Zavriev
  • E. V. Svirshchevskaya
Methods

Abstract

Successful disease prevention and therapy critically depend on timely diagnosis of infections. Quantitative immuno-PCR (qiPCR) technology improves the sensitivity in the detection of antibodies to pathogens. A qiPCR-based assay was developed to determine IgG antibodies to Epstein–Barr virus (EBV) in the human blood serum. EBV nuclear protein 1 fragment (pEBV) was expressed in Escherichia coli. A synthetic single-stranded deoxyribonucleotide was conjugated to streptavidin, and the conjugate was used to detect рEBV–IgG1–biotin complexes by qiPCR. The IgG1 titers determined by qiPCR were compared to the results of enzyme-linked immunosorbent assay (ELISA). The sensitivity of qiPCR was one order of magnitude higher than that of ELISA. Thus, a highly sensitive qiPCR-based assay was developed to quantitate antibodies specific to the recombinant EBV antigen.

Keywords

quantitative immuno-PCR ELISA recombinant proteins Epstein–Barr virus EBNA1 IgG1 

Abbreviations

ELISA

enzyme-linked immunosorbent assay

EBNA1

Epstein–Barr nuclear antigen 1

EBV

Epstein–Barr virus

IgG1

immunoglobulin G1

iPCR

immuno-PCR

qiPCR

quantitative iPCR

sODN

streptavidin–oligodeoxyribonucleotide complex

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Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  • V. D. Pivovarov
    • 1
  • D. Yu. Ryazantsev
    • 1
  • M. A. Simonova
    • 1
  • T. V. Yegorova
    • 1
  • S. V. Khlgatian
    • 2
  • S. K. Zavriev
    • 1
  • E. V. Svirshchevskaya
    • 1
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Mechnikov Institute of Vaccines and SeraMoscowRussia

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