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VETERINARY IMMUNOLOGY

Enzyme-Linked Immunosorbent Assay Based on a Chimeric Antigen Bearing Antigenic Regions of Structural Proteins Erns and E2 for Serodiagnosis of Classical Swine Fever Virus Infection

Min Lin, Erin Trottier, Maria Mallory
Min Lin
Animal Diseases Research Institute, Ottawa, Ontario, Canada K2H 8P9
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  • For correspondence: linm@inspection.gc.ca
Erin Trottier
Animal Diseases Research Institute, Ottawa, Ontario, Canada K2H 8P9
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Maria Mallory
Animal Diseases Research Institute, Ottawa, Ontario, Canada K2H 8P9
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DOI: 10.1128/CDLI.12.7.877-881.2005
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    FIG. 1.

    SDS-PAGE and Western blot analysis of the C21ErnsE2 chimeric protein expressed from pET184-177. The protein bands were stained with Coomassie blue (a) or immunostained with anti-His MAb (b), anti-Erns M2165 (c), anti-E2 M1655 (d), or nonspecific MAb M1875 (e). Lanes: M, the protein standards with their molecular masses in kilodaltons shown on the left; 1, whole-cell lysates of an E. coli clone (equivalent to 1 ml of culture with an A590 of 0.2) harboring pET184-177 in the absence of IPTG (isopropyl-β-d-thiogalactopyranoside); 2, whole-cell lysates of an E. coli clone harboring pET184-177 after induction with IPTG for 3 h; 3, nickel chelate affinity-purified C21ErnsE2 (5 μg in panel a, 1 μg in all other panels).

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    FIG. 2.

    Detection by ELISA of antibodies in sera of pigs experimentally infected with CSFV. Twenty serum samples from CSFV-infected pigs (Table 1) and 238 CSFV-negative pig serum samples were analyzed for reactivity with immobilized ELISA antigens Ernsaa 109-160 (a), E2AB (b), and C21ErnsE2 (c) at 2 μg/ml each. Dilution buffer without testing serum served as a control. The optimum cutoff determined by ROC analysis is indicated by a broken line. The locations of the Erns and E2 fragments relative to the full-length proteins are boxed in the insets. The three overlapping antigenic regions of Erns identified previously (24) are marked as AR1, AR2, and AR3.

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  • TABLE 1.

    Sera from pigs experimentally infected with various CSFV strains

    Serum sample (collection date)adpidpcbCSFV strain
    HC hyperimmune serum P15-93 (302-08) (9-15-94)15NAc
    HC hyperimmune serum P16-93 (9-21-94)21NA
    HC antiserum P43-83 (8-16-83)17NA
    HC standard serum P78-82 (4-18-83)166NA
    HC P12-93 (1-12-94)8Alfort
    HC P17-93 (12-29-93)56Glentorf
    HC P18-93 (12-29-93)56Glentorf
    HC P19-93 (12-30-93)57Glentorf
    HC P4-92 (6-2-92)7Glentorf
    HC P4-92 (6-30-92)35Glentorf
    HC P5-92 (6-2-92)7Glentorf
    HC P5-92 (6-9-92)14Glentorf
    HC P5-92 (6-16-92)21Glentorf
    HC P5-92 (6-30-92)35Glentorf
    HCV hyperimmune serum P222 (6-2-64)NANA
    HCV antiserum L-1 P154 (6-2-66)210Lapinized
    HCV antisera NT P155 (6-22-66)210Lapinized
    HCV immune sera 51PIC/41 P2427 P3C (1-16-69)71NA
    HCV antiserum P54-83 (10-24-83)NANA
    HC PA03 (12-14-98)7Alfort
    • ↵ a Collection dates shown are month-day-year. HC, hog cholera; HCV, hog cholera virus.

    • ↵ b dpc, days postchallenge. These animals were infected with one CSFV strain and then challenged with other CSFV strains during the course of infection.

    • ↵ c NA, not available.

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Enzyme-Linked Immunosorbent Assay Based on a Chimeric Antigen Bearing Antigenic Regions of Structural Proteins Erns and E2 for Serodiagnosis of Classical Swine Fever Virus Infection
Min Lin, Erin Trottier, Maria Mallory
Clinical and Diagnostic Laboratory Immunology Jul 2005, 12 (7) 877-881; DOI: 10.1128/CDLI.12.7.877-881.2005

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Enzyme-Linked Immunosorbent Assay Based on a Chimeric Antigen Bearing Antigenic Regions of Structural Proteins Erns and E2 for Serodiagnosis of Classical Swine Fever Virus Infection
Min Lin, Erin Trottier, Maria Mallory
Clinical and Diagnostic Laboratory Immunology Jul 2005, 12 (7) 877-881; DOI: 10.1128/CDLI.12.7.877-881.2005
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