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Detection of staphylococcal enterotoxin a by phage display mediated immuno-PCR method

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Abstract

A method of staphylococcal enterotoxin A (SEA) detection in pasteurized milk using recombinant miniantibodies exposed on phage particles was developed. Miniantibodies bind to the antigen, while phage DNA serves as a matrix in polymerase chain reaction (PCR). The SEA detection limit in milk by a phage display mediated immuno-PCR method is 100 pg/mL that is much lower than the level of toxic and allergic dose.

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Abbreviations

SEA:

staphylococcal enterotoxin A

BSA:

bovine serum albumin

PBS:

phosphate buffered saline (pH 7.4)

PBST:

PBS with 0.1% Tween 20

PBSTM:

PBST with 1% skimmed milk powder

scFv:

single-chain mini-antibodies

ELISA:

enzyme-linked immunosorbent assay

TAE:

Tris-acetate buffer

PCR:

polymerase chain reaction

iPCR:

immuno- PCR

mAbSEA:

monoclonal mouse antibodies against SEA

scFv5:

monoclonal single-chain mini-antibodies against SEA

p.p.:

phage particle.

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Authors and Affiliations

  1. Ernst All-Russia Research Institute of Animal Husbandry, Dubrovitsy, Moscow oblast, 142132, Russia

    K. K. Fursova, M. P. Shchannikova, I. V. Loskutova, A. O. Shepelyakovskaya & F. A. Brovko

  2. Branch of the Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    A. A. Artykov, K. K. Fursova, M. P. Shchannikova, I. V. Loskutova, A. O. Shepelyakovskaya, A. G. Laman & F. A. Brovko

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

    D. Yu. Ryazantsev & S. K. Zavriev

Authors
  1. A. A. Artykov
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  2. K. K. Fursova
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  3. D. Yu. Ryazantsev
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  4. M. P. Shchannikova
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  5. I. V. Loskutova
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  6. A. O. Shepelyakovskaya
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  7. A. G. Laman
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  8. S. K. Zavriev
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  9. F. A. Brovko
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Corresponding author

Correspondence to F. A. Brovko.

Additional information

Original Russian Text © A.A. Artykov, K.K. Fursova, D.Yu. Ryazantsev, M.P. Shchannikova, I.V. Loskutova, A.O. Shepelyakovskaya, A.G. Laman, S.K. Zavriev, F.A. Brovko, 2017, published in Bioorganicheskaya Khimiya, 2017, Vol. 43, No. 5, pp. 518–522.

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Artykov, A.A., Fursova, K.K., Ryazantsev, D.Y. et al. Detection of staphylococcal enterotoxin a by phage display mediated immuno-PCR method. Russ J Bioorg Chem 43, 540–543 (2017). https://doi.org/10.1134/S1068162017050028

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  • DOI: https://doi.org/10.1134/S1068162017050028

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