Key result
The Rec p30-IB immunoblotting test system for African swine fever demonstrated a diagnostic sensitivity of 100.00% and specificity of 98.75% in samples from domestic pigs and wild boars.
The developed immunoblotting test system using recombinant p30 demonstrated high sensitivity and specificity for detecting African swine fever virus antibodies in pigs and wild boars.
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May support ASF surveillance in swine; leaves open prospective field validation before routine adoption.
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).
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Kazakova et al. (2016) studied African swine fever (ASF). Immunoblotting test system (Rec p30-IB) was evaluated on Diagnostic specificity and sensitivity (Sensitivity 100.00%, Specificity 98.75%). The Rec p30-IB immunoblotting test system for African swine fever demonstrated a diagnostic sensitivity of 100.00% and specificity of 98.75% in samples from domestic pigs and wild boars.
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