Application of Competitive Enzyme-Linked Immunosorbent Assay for the Serologic Diagnosis of Classical Swine Fever Virus Infection

A competitive enzyme-linked immunosorbent assay (C-ELISA), based on a truncated E2 recombinant protein of the Alfort/187 strain of classical swine fever virus (CSFV) and a specific monoclonal antibody M1669, was evaluated using 2,000 sera from clinically healthy pigs in Canada (a CSFV-free country) and sera from experimentally infected pigs. The relative specificity and sensitivity of the C-ELISA were 100% and 86%, respectively, at a cutoff of 25% inhibition using negative and positive pig sera, as defined by the neutralizing peroxidase-linked assay (NPLA). A kappa value of 0.91 was obtained, indicating an excellent level of agreement between the NPLA and the C-ELISA. When sera from 120 infected pigs were used in the test at ≥21 days postinfection, the sensitivity of the C-ELISA and the kappa value increased to 97% and 0.98, respectively. This C-ELISA will be useful when a large number of samples must be tested, as could occur during a disease outbreak or for surveillance or prevalence studies.

[1]  A. Ressang,et al.  The indirect fluorescent antibody technique as a method for detecting serum antibodies against hog cholera. I. An outline of the technique and its preliminary evaluation. , 2010, Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B.

[2]  A. Bouma,et al.  Laboratory decision-making during the classical swine fever epidemic of 1997-1998 in The Netherlands. , 1999, Preventive veterinary medicine.

[3]  A. Dijkhuizen The 1997–1998 outbreak of classical swine fever in The Netherlands , 1999 .

[4]  A A Dijkhuizen,et al.  A model to estimate the financial consequences of classical swine fever outbreaks: principles and outcomes. , 1999, Preventive veterinary medicine.

[5]  M. D. de Jong,et al.  Transmission of classical swine fever virus within herds during the 1997-1998 epidemic in The Netherlands. , 1999, Preventive veterinary medicine.

[6]  P. V. van Rijn,et al.  An experimental marker vaccine and accompanying serological diagnostic test both based on envelope glycoprotein E2 of classical swine fever virus (CSFV). , 1999, Vaccine.

[7]  G. Wensvoort,et al.  An improved ELISA for the detection of serum antibodies directed against classical swine fever virus. , 1997, Veterinary microbiology.

[8]  K. Depner,et al.  Evaluation of a gp 55 (E2) recombinant-based ELISA for the detection of antibodies induced by classical swine fever virus. , 1996, DTW. Deutsche tierarztliche Wochenschrift.

[9]  J. Tratschin,et al.  Detection of antibodies against classical swine fever virus in swine sera by indirect ELISA using recombinant envelope glycoprotein E2. , 1996, Veterinary microbiology.

[10]  M. Harding,et al.  Reverse transcriptase-PCR assay for detection of hog cholera virus , 1994, Journal of clinical microbiology.

[11]  P. Wright,et al.  Comparison of competitive and indirect enzyme-linked immunosorbent assays for detection of bluetongue virus antibodies in serum and whole blood , 1987, Journal of clinical microbiology.

[12]  A. Gielkens,et al.  The neutralizing peroxidase-linked assay for detection of antibody against swine fever virus. , 1984, Veterinary microbiology.

[13]  J. Hanley,et al.  The meaning and use of the area under a receiver operating characteristic (ROC) curve. , 1982, Radiology.

[14]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. Köhler,et al.  A better cell line for making hybridomas secreting specific antibodies , 1978, Nature.

[16]  C. Metz Basic principles of ROC analysis. , 1978, Seminars in nuclear medicine.

[17]  H. Zola,et al.  Monoclonal antibodies : preparation and use of monoclonal antibodies and engineered antibody derivatives , 2000 .

[18]  P. Have Detection of antibodies against swine fever virus by enzyme-linked immunosorbent assay (ELISA). , 1984, Acta veterinaria Scandinavica.