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Clinical and Vaccine Immunology, July 2009, p. 1012-1020, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00408-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Recombinant Antigen Targets for Serodiagnosis of African Swine Fever {triangledown}

Carmina Gallardo,1,6 Ana Luísa Reis,2 Gladys Kalema-Zikusoka,3 Joana Malta,4 Alejandro Soler,1 Esther Blanco,1 R. M. E. Parkhouse,2* and Alexandre Leitão5*

Centro de Investigación en Sanidad Animal, INIA, Valdeolmos, Madrid 28130, Spain,1 Instituto Gulbenkian de Ciência, Apartado 14, Oeiras 2781-901, Portugal,2 Conservation through Public Health, P.O. Box 10950, Kampala, Uganda,3 CesNova, Faculdade de Ciências Sociais e Humanas, Universidade Nova de Lisboa, Av. Berna 26C, Lisbon 1069-061, Portugal,4 Instituto de Investigação Científica Tropical, CVZ, CIISA, Av. Universidade Técnica, Lisbon 1300-477, Portugal,5 International Livestock Research Institute, P.O. Box 30709, Kabete, Nairobi, Kenya6

Received 7 November 2008/ Returned for modification 22 December 2008/ Accepted 7 April 2009

African swine fever (ASF) is an infectious and economically important disease of domestic pigs. There is no vaccine, and so reliable diagnosis is essential for control strategies. The performance of four recombinant ASF virus (ASFV) protein (pK205R, pB602L, p104R, and p54)-based enzyme-linked immunosorbent assays (ELISAs) was evaluated with European porcine field sera that had been established by Office International des Epizooties (OIE)-approved tests to be ASFV negative (n = 119) and ASFV positive (n = 80). The {kappa} values showed that there was almost perfect agreement between the results of the "gold standard" test (immunoblotting) and the results obtained by the p54-specific ELISA ({kappa} = 0.95; 95% confidence interval [CI], 0.90 to 0.99) and the pK205R-specific ELISA or the pB602L-specific ELISA ({kappa} = 0.92; 95% CI, 0.86 to 0.97). For the pA104R-specific ELISA, there was substantial to almost perfect agreement ({kappa} = 0.81; 95% CI, 0.72 to 0.89). Similar results were observed by the OIE-approved ELISA ({kappa} = 0.89; 95% CI, 0.82 to 0.95). Importantly, antibodies against these proteins were detectable early after infection of domestic pigs. Preliminary testing of 9 positive and 17 negative serum samples from pigs from West Africa showed identical results by the recombinant protein-based ELISA and the OIE-approved tests. In contrast, there was a high degree of specificity but a surprisingly a low level of sensitivity with 7 positive and 342 negative serum samples from pigs from East Africa. With poorly preserved sera, only the p104R-specific ELISA showed a significant reduction in sensitivity compared to that of the OIE-approved ELISA. Finally, these recombinant proteins also detected antibodies in the sera of the majority of infected warthogs. Thus, recombinant ASFV proteins p54, pB602L, and pK205R provide sensitive and specific targets for the detection of antibodies in European and West African domestic pigs and warthogs.

* Corresponding author. Mailing address for R. M. E. Parkhouse: Instituto Gulbenkian de Ciência, Apartado 14, Oeiras 2781-901, Portugal. Phone: 351 214 407 939. Fax: 351 214 407 970. E-mail: parkhous{at}igc.gulbenkian.pt. Mailing address for Alexandre Leitão: Instituto de Investigação Científica Tropical, CVZ, CIISA, Av, Universidade Técnica, Lisbon 1300-477, Portugal. Phone: 351 213 652 800. Fax: 351 213 652 869. E-mail: alexandre{at}fmv.utl.pt

{triangledown} Published ahead of print on 6 May 2009.

Clinical and Vaccine Immunology, July 2009, p. 1012-1020, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00408-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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