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

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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.

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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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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia

    V. D. Pivovarov, D. Yu. Ryazantsev, M. A. Simonova, T. V. Yegorova, S. K. Zavriev & E. V. Svirshchevskaya

  2. Mechnikov Institute of Vaccines and Sera, Moscow, 105064, Russia

    S. V. Khlgatian

Authors
  1. V. D. Pivovarov
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  2. D. Yu. Ryazantsev
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  3. M. A. Simonova
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  4. T. V. Yegorova
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  5. S. V. Khlgatian
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  6. S. K. Zavriev
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  7. E. V. Svirshchevskaya
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Corresponding author

Correspondence to E. V. Svirshchevskaya.

Additional information

Original Russian Text © V.D. Pivovarov, D.Yu. Ryazantsev, M.A. Simonova, T.V. Yegorova, S.V. Khlgatian, S.K. Zavriev, E.V. Svirshchevskaya, 2018, published in Molekulyarnaya Biologiya, 2018, Vol. 52, No. 4, pp. 727–734.

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Pivovarov, V.D., Ryazantsev, D.Y., Simonova, M.A. et al. Immuno-PCR Assay for Quantitation of Antibodies to Epstein–Barr Virus. Mol Biol 52, 629–635 (2018). https://doi.org/10.1134/S0026893318040155

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  • DOI: https://doi.org/10.1134/S0026893318040155

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