title: Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay.
abstract: 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).
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: A S
authors.initials: AS
authors.lastname: Kazakova
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: I R
authors.initials: IR
authors.lastname: Imatdinov
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: O A
authors.initials: OA
authors.lastname: Dubrovskaya
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: A R
authors.initials: AR
authors.lastname: Imatdinov
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: M V
authors.initials: MV
authors.lastname: Sidlik
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: V M
authors.initials: VM
authors.lastname: Balyshev
authors.affiliation: Republican Scientific-Research Subsidiary Unitary Enterprise 'The S.N. Vyshelessky Institute of Experimental Veterinary', Minsk, Belarus.
authors.forename: P A
authors.initials: PA
authors.lastname: Krasochko
authors.affiliation: National Research Institute of Veterinary Virology and Microbiology of Russia (VNIIVViM), Volginsky, Petushki Area, Vladimir Region, Russia.
authors.forename: A D
authors.initials: AD
authors.lastname: Sereda
mesh_terms: D000357:African Swine Fever / Q000175:diagnosis*; D000358:African Swine Fever Virus / Q000302:isolation & purification*; D000818:Animals; D015151:Immunoblotting / Q000379:methods / Q000662:veterinary*; D010750:Phosphoproteins / Q000097:blood*; D011994:Recombinant Proteins / Q000097:blood; D034421:Sus scrofa; D013552:Swine; D014764:Viral Proteins / Q000097:blood*
chemical_list: D010750:Phosphoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C074960:P30 protein, African Swine Fever Virus
keywords: African swine fever; antibody detection; immunoblotting; p30; recombinant protein; serological diagnosis