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Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA.

In the absence of an effective vaccine (s), the control of African swine fever is caused by African swine fever virus (ASFV) should be based on early, efficient, cost-effective detection and control and elimination of strict strategy. To this end, we developed an indirect ELISA was able to detect antibodies ASFV either serum or oral fluid specimens. 

Recombinant proteins used in the ELISA selected by comparing the serum antibody responses beginning of ASFV-infected pigs (NHV-P68 isolates) Fruit fly Recombinant Proteins for the three major recombinant polypeptide (p30, P54, p72) using multiplex fluorescent microbead-based immunoassay (FMIA). Not dangerous serum (non-infectious) antibody-positive to be used as a positive control plates and for the calculation of sample-to-positive (S: P) ratio was produced by inoculation of pigs with replicon particle vaccines (RP) revealed that ASFV p30 gene.

 Optimized ELISA detected anti-p30 antibodies in serum and / or oral fluid samples from pigs inoculated with ASFV under experimental conditions ranging from 8 to 12 days post inoculation. Tests in serum (n = 200) and oral fluid (n = 200) samples the field of free ASFV population indicates that the test is highly specific diagnostic. 

Comfort and utility of diagnostic sampling oral fluid combined with the flexibility to test either serum or oral fluid on the same platform shows that this test will be very useful in the condition that the OIE recommends monitoring antibody ASFV, namely, in the area of ​​ASFV-endemic and for detecting infection by ASFV low virulence isolates.


Recombinant Protein p30 for serological diagnosis of African Swine Fever by Immunobloiting Assay.


This article is devoted to the development and evaluation of immunoblotting test systems for the serological diagnosis of African swine fever (ASF), based on highly purified recombinant p30 of ASF virus (ASFV) strain 01/08 Stavropol (Stavropol, 2008), the representative of ASFV currently circulating in Russian Federation. 

The main project phases are as follows: (i) the cloning of the central hydrophilic region CP204L ASFV gene (p30) into a prokaryotic vector; (Ii) the expression of p30 and chromatographic purification of recombinant products with thioredoxin and poly-histidine site (p30e1_TrxA_6xHis); (Iii) the development of test systems immunoblotting (Rec p30-IB) using highly purified recombinant p30; and (iv) the
Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA.
evaluation of p30-IB Rec using General Recombinant Proteins  sera and organ samples from domestic pigs and wild boar experimentally or naturally infected by ASFV. 

Rec Testing p30-IB show diagnostic specificity and sensitivity of the test to be 98.75% and 100.00%, respectively. high sensitivity of Rec p30-IB allowed the detection of antibodies ASFV specific in samples of organs of the immune system and sera of blood, collected from domestic pigs and wild boar, from 6 to 8 days post-infection, regardless of virus virulence, seroimmunotype and origin geographic sample
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