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A. Kittawornrat 1, J. Prickett 1, C. Wang 1,2, C. Olsen 1, C. Irwin 1, Y. Panyasing 1, A. Ballagi 3, A. Rice 3, R. Main 1, C. Rademacher 4, M. Hoogland.

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Presentation on theme: "A. Kittawornrat 1, J. Prickett 1, C. Wang 1,2, C. Olsen 1, C. Irwin 1, Y. Panyasing 1, A. Ballagi 3, A. Rice 3, R. Main 1, C. Rademacher 4, M. Hoogland."— Presentation transcript:

1 A. Kittawornrat 1, J. Prickett 1, C. Wang 1,2, C. Olsen 1, C. Irwin 1, Y. Panyasing 1, A. Ballagi 3, A. Rice 3, R. Main 1, C. Rademacher 4, M. Hoogland 4, J. Lowe 5, J. Zimmerman 1 1 Department of Veterinary Diagnostic and Production Animal Medicine, Iowa State University, Ames, IA 2 Department of Statistics, College of Liberal Arts and Sciences, Iowa State University, Ames, IA 3 IDEXX Laboratories, Inc., Westbrook, ME 4 Murphy-Brown LLC, Ames, IA 5 Carthage Veterinary Service, Ltd., Carthage IL

2 In human diagnostic medicine 1909. “The agglutinating properties of several body fluids during Malta Fever.” Pollaci and Ceralo 1986. Oral fluid antibodies used to detect HIV infection. J Clin Microbiol 24(5): 873 Lots of research in humans (1909- 2010) Primary focus on antibody detection Widely used in surveillance

3 BOARS: Confirm negative PRRS status of individual “seed stock” pigs GILTS: Confirm PRRS status before entering breeding herds, preferably at the beginning and end of quarantines SOWS and PIGLETS: Monitor vaccine effectiveness or confirm PRRS status Boars Gilts Sows Piglets Simplified sampling would allow more frequent testing and provide more exact epidemiological information

4 Pictures courtesy of Dr. John Prickett

5

6 1. Optimize antibody isotypes 2. Optimize incubation period 3. Optimize sample dilution 4. Interactions (isotype x sample dilution x sample volume) 5. Optimize conjugate dilution

7 Conclusion from previous results

8 Serum antibody assays (IDEXX PRRS X3 can be optimized to detect antibodies in oral fluid by -IgG FC isotype (IgM, IgA, IgG can be detected) -250 μ l sample volume -16 hour incubation -1:2 sample dilution -Other conjugate

9 Wean-to-finish barns (1100 pigs) 10 sites x 6 pens per site x sampling each 2 weeks. n = 600 total samples 10 barns: 1,100 head wean-finish - 1 oral fluid sample / pen; 6 pens /barn -Samples collected every 2 weeks from placement to close-out (10 sample points) -600 oral fluid samples (PCR, ELISA)

10 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48

11 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P Avg. S/P = 0.30 Pos. = 23.3% Avg. S/P = 0.04 Pos. = 3.3% Avg. S/P = 0.72 Pos. = 53.3% Avg. S/P = 0.34 Pos. = 30.0% 5 serum samples per pen (n = 30) at week 0

12 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

13 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

14 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

15 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

16 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

17 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

18 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

19 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P

20 WEEK POST PLACEMENT BarnPen024681012141618 512.470.690.180.00 -0.030.000.01 P 2.111.131.68 24.660.980.180.120.05 -0.010.01 P 2.012.660.96 33.380.700.140.03 -0.03 -0.040.13 P 3.122.750.82 4NS1.280.140.180.030.010.01 P 2.732.480.98 51.251.870.270.12 -0.01 -0.03 -0.03 P 2.083.272.11 62.730.960.070.020.01 -0.03 -0.012.291.381.13 610.831.290.00 -0.02 -0.042.292.461.981.79 20.040.51 -0.02 0.14 -0.022.41 P 2.962.311.40 30.001.13 -0.04 2.692.762.190.47 40.080.620.00 -0.03 -0.04 -0.032.93 P 3.052.120.89 50.000.79 -0.02 -0.01 -0.020.03 -0.022.18 P 1.721.46 6 -0.010.040.030.060.010.192.272.991.310.37 914.151.720.210.170.260.06 -0.020.010.023.20 24.271.790.340.190.07 -0.020.020.000.301.28 34.411.760.29 -0.020.080.020.00 -0.033.12 43.291.590.270.100.05 -0.020.03 -0.010.022.49 54.021.430.510.050.000.01 -0.010.040.023.28 64.010.910.830.380.130.030.060.020.012.19 P 101 -0.030.260.210.49 P 1.621.652.661.602.100.89 23.650.750.11 P 1.46 P 1.001.243.270.932.261.92 33.790.260.48 P 2.031.172.502.742.210.561.31 44.390.550.020.24 P 2.42 P 2.272.871.942.231.45 53.690.060.010.562.111.723.212.311.793.29 63.710.080.090.00 P 3.162.133.281.251.440.48 < 0.40 ≥ 0.40 ≥ 0.40 P < 0.40 P NS P Avg. S/P = 0.93 Pos. = 93.3% Avg. S/P = 0.78 Pos. = 70.0% Avg. S/P = 1.40 Pos. = 96.7% Avg. S/P = 1.06 Pos. = 86.7% 30 serum samples at week 18

21 Source of positive samples – Experimental samples – Field samples – For any pen, all samples after the first PCR positive result were considered positive Source of negative samples – Experimental (DPI 0) – Field samples from expected PRRSV negative herds submitted to ISU VDL

22 NEGATIVE SAMPLES Experimental= 84 Field neg (ISU VDL)= 283 TOTAL= 367 POSITIVE SAMPLES Experimental= 251 Field= 241 TOTAL= 492

23

24 Kinetics of anti-PRRSV antibodies in oral fluid is amenable to rapid detection of infection. The oral fluid ELISA could provide an efficient, cost- effective approach to PRRSV monitoring in commercial herds and surveillance in elimination programs.

25 IDEXX Laboratories, Inc. National Pork Board (Pork Checkoff funds) PRRS CAP, USDA NIFA award 2008-55620-1932 Request reference standard samples contact to Dr. Zimmerman (jjzimm@iastate.edu)jjzimm@iastate.edu Request SOP contact to Dr. Johnson (jkjohn@iastate.edu)jkjohn@iastate.edu

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