Clinical Veterinary Microbiology

Detection of U.S., Lelystad, and European-Like Porcine Reproductive and Respiratory Syndrome Viruses and Relative Quantitation in Boar Semen and Serum Samples by Real-Time PCR

A. Wasilk, J. D. Callahan, J. Christopher-Hennings, T. A. Gay, Y. Fang, M. Dammen, M. E. Reos, M. Torremorell, D. Polson, M. Mellencamp, E. Nelson, W. M. Nelson
DOI: 10.1128/JCM.42.10.4453-4461.2004

Article Figures & Data

Figures

  • FIG. 1.
    FIG. 1.

    Standard curve of number of RNA copies per milliliter (dashed line) and 12 replicates of each concentration (3 to 6 replicates per day for 3 days) (solid line) versus the mean Ct (y = −0.31x + 11.441). Bars indicate standard errors.

  • FIG. 2.
    FIG. 2.

    Viral load in serum, in TCID50s per milliliter, as determined by virus titration (▪), and in numbers of copies per milliliter, as determined by quantitative real-time PCR (□). Values are for the first 10 DPI for each PRRSV-inoculated boar.

  • FIG. 3.
    FIG. 3.

    Viral loads (number of RNA copies per milliliter) and viremia and shedding patterns in PRRSV-inoculated boars. ▪, detection of PRRSV, quantitation in semen, and shedding patterns; ⧫, PRRSV detection and quantitation in serum.

Tables

  • TABLE 1.

    Viral loads in serum and semen samples from PRRSV-inoculated boars

    Boar no. and sampleViral load (no. of RNA copies/ml) on the following DPI:
    135810121517192224262932363944515867727986
    1
        Serum9 × 1012 × 104a3 × 104a4 × 1045 × 1011 × 102b0002 × 10304 × 103002 × 102b00000000
        Semen0004 × 1043 × 104000000000000000000
    2
        Serum2 × 1025 × 104a2 × 105a1 × 1054 × 104a7 × 1023 × 1021 × 1032 × 1030Nc00003 × 101b000000
        Semen00000000000000000000000
    3
        Serum2 × 103a8 × 104a2 × 105a3 × 104a8 × 1017 × 1021 × 1026 × 11b5 × 1011 × 102bNN6 × 1030000000000
        Semen06 × 106000000000000000000000
    4
        Serum3 × 1036 × 104a1 × 105a5 × 1035 × 101b6 × 1033 × 1020N2 × 1020004 × 1029 × 101b0000008 × 101b0
        Semen0006 × 1022 × 101b000000000000000000
    5
        Serum2 × 103a8 × 104a9 × 105a4 × 1053 × 1032 × 104a5 × 1022 × 1025 × 1039 × 102N3 × 102006 × 101b00000000
        Semen009 × 10409 × 10100NN000NDd0000000000
    6
        Serum3 × 103a5 × 104a2 × 105a5 × 104a7 × 1026 × 101N4 × 102N6 × 101bN3 × 1020N000000000
        Semen001 × 1061 × 1052 × 1068 × 1053 × 1043 × 10502 × 1061 × 10406 × 1033 × 103000000000

    • ↵a PRRSV was detected by virus isolation.

    • ↵b nPCR negative, real-time PCR positive.

    • ↵c N, nPCR positive, real-time PCR negative.

    • ↵d ND, no sample was collected.

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