Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay.

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 CP204L (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).

[Continuous cell subline A4C2/9K and its application to the african swine fever virus study].

A new continuous cell subline A4C2/9K highly sensitive to the african swine fever virus (ASFV) was prepared. All the tested ASFV strains isolated in the Russian Federation in 2008-2013 proliferated in this cell culture exhibiting hemadsorption and accumulated at a titer of up to 6.5 Ig HAU50/cm3. The cell culture A4C2/9K can be used for ASFV isolation or determination of its infectious activity and serotype identity. The culture versions of the ASFV strain Stavropol 01/08 at passages 24 and 33 in the cell culture A4C2/9K lost their pathogenicity for pigs.

[Antigenic diversity of African swine fever viruses].

Data on the seroimmunotypic and hemadsorbing characteristics of African swine fever virus (ASF) are summarized. According to the results of immunological sampling in pigs and those of hemagglutination inhibition test, the known ASFV strains and isolates were divided into 11 groups, 8 were characterized as seroimmunogroups having their specific reference strains. A 110-140-kD ASFV serotype-specific nonstructural major glycoprotein was identified. It is suggested that it is the glycoprotein that corresponds to the genetic engineering detected virus-specific homolog of lymphocyte membrane protein CD2, gene deletion of which results in the loss of hemadsorbing properties by ASFV.