Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Sep 9;11(9):e0161230.
doi: 10.1371/journal.pone.0161230. eCollection 2016.

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

Luis G Giménez-Lirola  1 , Lina Mur  2 , Belen Rivera  2 , Mark Mogler  3 , Yaxuan Sun  4 , Sergio Lizano  5 , Christa Goodell  5 , D L Hank Harris  3 , Raymond R R Rowland  6 , Carmina Gallardo  7 , José Manuel Sánchez-Vizcaíno  2 , Jeff Zimmerman  1
Affiliations

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

Luis G Giménez-Lirola et al. PLoS One. .

Abstract

In the absence of effective vaccine(s), control of African swine fever caused by African swine fever virus (ASFV) must be based on early, efficient, cost-effective detection and strict control and elimination strategies. For this purpose, we developed an indirect ELISA capable of detecting ASFV antibodies in either serum or oral fluid specimens. The recombinant protein used in the ELISA was selected by comparing the early serum antibody response of ASFV-infected pigs (NHV-p68 isolate) to three major recombinant polypeptides (p30, p54, p72) using a multiplex fluorescent microbead-based immunoassay (FMIA). Non-hazardous (non-infectious) antibody-positive serum for use as plate positive controls and for the calculation of sample-to-positive (S:P) ratios was produced by inoculating pigs with a replicon particle (RP) vaccine expressing the ASFV p30 gene. The optimized ELISA detected anti-p30 antibodies in serum and/or oral fluid samples from pigs inoculated with ASFV under experimental conditions beginning 8 to 12 days post inoculation. Tests on serum (n = 200) and oral fluid (n = 200) field samples from an ASFV-free population demonstrated that the assay was highly diagnostically specific. The convenience and diagnostic utility of oral fluid sampling combined with the flexibility to test either serum or oral fluid on the same platform suggests that this assay will be highly useful under the conditions for which OIE recommends ASFV antibody surveillance, i.e., in ASFV-endemic areas and for the detection of infections with ASFV isolates of low virulence.

PubMed Disclaimer

Conflict of interest statement

The authors declare the following potential conflicts of interest with respect to the research authorship, and/or publication of this article: authors S. Lizano and C. Goodell are employed by IDEXX Laboratories, Inc. The remaining authors declare no conflicting interests with respect to their authorship or the publication of this article. The commercial affiliation above mentioned does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. ASFV multiplex fluorescent microbead-based immunoassay (FMIA) sample-to-positive (S/P) serum antibody response (mean, SE) against three recombinant antigens (p30, p54, p72) in 9 pigs inoculated with ASFV NHV/P68 (experiment 1).
Fig 2
Fig 2
ASFV recombinant p30 antibody ELISA sample-to-positive (S/P) responses in (2A) individual pig serum (mean, SE) and (2B) pen-based oral fluid samples following inoculation with replicon particle (RP) vaccines expressing ASFV p30 or a combination of ASFV p30/54/72. Controls were defined as unvaccinated pigs (one pen of 4 pigs) and pigs inoculated with RP vaccines expressing ASFV p54 (one pen of 8 pigs) or ASFV p72 (one pen of 8 pigs). In all cases, pigs received two doses of vaccine (days 0 and 21).
Fig 3
Fig 3. ASFV recombinant p30 antibody ELISA serum-to-positive (S/P) responses in serum (mean, SE) and oral fluid (mean, SE) samples from 17 pigs (experiments 1 and 2) inoculated with ASFV NHV/P68.
Fig 4
Fig 4
Distribution of ASFV recombinant p30 antibody ELISA serum-to-positive (S/P) responses in (A) serum (n = 200) and (B) oral fluid (n = 200) specimens from North American commercial pigs and (C) serum (n = 52) and (D) oral fluid (n = 46) collected ≥ 14 days following inoculation with ASFV NHV/P68.
See this image and copyright information in PMC

References

    1. Dixon LK, Escribano JM, Martins C, Rock DL, Salas ML, Wilkinson PJ. Asfarviridae In: Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA, editors. Virus Taxonomy. VIII Report of the ICTV. London: Elsevier, Academic Press; 2005. pp. 135–143.
    1. Oura CAL, Arias M. African swine fever; 2012. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. OIE, World Organisation for Animal Health. Available: http://www.oie.int/international-standard-setting/terrestrial-manual/acc.... Accessed 26 May 2015.
    1. Sánchez-Vizcaíno JM, Mur L, Gomez-Villamandos JC, Carrasco L. An update on the epidemiology and pathology of African swine fever. J Comp Path. 2015; 152: 9–21. 10.1016/j.jcpa.2014.09.003 - DOI - PubMed
    1. Arias M, Sánchez-Vizcaíno JM. African swine fever In: Zimmerman JJ, Karriker LA, Ramirez A, Schwartz KJ, Stevenson GW, editors. 10th ed. Diseases of Swine. Hoboken, New Jersey: Wiley-Blackwell; 2012. pp. 396–404.
    1. European Food Safety Authority (EFSA). Evaluation of possible mitigation measures to prevent introduction and spread of African swine fever virus through wild boar. EFSA Journal. 2014; 12(3): 3616.

MeSH terms

LinkOut - more resources