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Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay
Author(s) -
Kazakova A. S.,
Imatdinov I. R.,
Dubrovskaya O. A.,
Imatdinov A. R.,
Sidlik M. V.,
Balyshev V. M.,
Krasochko P. A.,
Sereda A. D.
Publication year - 2017
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.12539
Subject(s) - african swine fever virus , recombinant dna , serology , virology , biology , antibody , virus , cloning (programming) , microbiology and biotechnology , gene , immunology , biochemistry , computer science , programming language
Summary This article is devoted to the development and evaluation of the immunoblotting test system for serological diagnosis of African swine fever ( ASF ), based on the highly purified recombinant p30 of ASF virus ( ASFV ) strain Stavropol 01/08 (Stavropol 2008), representative of the ASFV currently circulating in the Russian Federation. The main project stages are as follows: (i) cloning of the central hydrophilic region of the ASFV gene CP 204L (p30) into a prokaryotic vector; (ii) expression and chromatographic purification of the recombinant product p30 with thioredoxin and poly‐histidine site (p30e1_TrxA_6xHis); (iii) development of the immunoblotting test system (Rec p30‐ IB ) using the highly purified recombinant p30; and (iv) evaluation of Rec p30‐ IB using sera and organ samples from domestic pigs and wild boars experimentally or naturally infected by ASFV . Testing of the Rec p30‐ IB showed the diagnostic specificity and sensitivity of the assay to be 98.75% and 100.00%, respectively. High sensitivity of the Rec p30‐ IB allowed the detection of ASFV ‐specific antibodies in samples of organs of the immune system and blood sera, collected from domestic pigs and wild boars, starting from 6 to 8 days post‐infection, regardless of virus virulence, seroimmunotype and geographic origin of the samples (East Europe, South Europe, West Europe, Central and south‐east Africa).