Summary of calves borne in both farms in North Rhine-Westphalia, Germany.

<p>The first number indicates the total number of calves in the respective group, inside the parentheses the number of calves is given separately for both holdings.</p

Detailed analyses of newborn calves in 2 farms in North Rhine-Westphalia.

<p>Meconium samples of calves whose ELISA results are marked in red tested positive by real-time RT-PCR; blue ELISA results correspond to negative PCR results. The day of gestation at the estimated time of SBV-infection of the respective cow is symbolized by a light blue area.</p

Sequences of positive standards used in this study, primer and probe binding regions are depicted in capitals.

<p>Sequences of positive standards used in this study, primer and probe binding regions are depicted in capitals.</p

Analytical sensitivity of the PRRSV-multiplex-assay based on 10-fold dilution series of positive standard RNA.

<p>Regression curve was estimated by Bio-Rad CFX Manager™ Software (Bio-Rad, Hercules, CA, USA).</p

Analyses of diagnostic sample from PRRSV infected swine with the multiplex RT-qPCR and with the ADIAVET® PRRS EU/NA Kit.

<p>Analyses of diagnostic sample from PRRSV infected swine with the multiplex RT-qPCR and with the ADIAVET® PRRS EU/NA Kit.</p

Histopathology, liver, mouse 5a.

<p>HE stain. Hepatocellular centroacinar degeneration (periportal, zone 1) and necrosis.</p

Chequerboard titration of a type 1- and a HP-PRRSV strain.

<p>Cq-values (z-axis) of the type 2- (2a) and HP- (2b) specific assays while co-amplification of different concentrations of the type 1 strain (y-axis) are shown. 2c depicts results of the type 1-specific assay when co-amplifying varying concentrations of HP-specific sequences (x-axis). Cq-values of the internal control RT-qPCR are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038251#pone-0038251-g002" target="_blank">figure 2d</a>. Images were designed supported by SigmaPlot® software.</p

Sequences of primers and probes used in the study.

*<p>genome position according to.</p><p>type 1-PRRSV: type 1 prototype strain Lelystad (accession number M96262).</p><p>type 2-PRRSV: type 2 prototype strain VR-2332 (accession number U87392).</p><p>HP-PRRSV: Porcine respiratory and reproductive syndrome virus strain JXwn06.</p><p>(accession number EF641008).</p><p>EGFP: cloning vector pEGFP-1 (accession number U55761).</p

Gross pathology, abdominal cavity, mouse 5a.

<p>Enlarged spleen and diffusely beige liver with multifocal hemorrhages.</p

Body weights of mice.

<p>Mice marked in red died spontaneously or had to be sacrificed prematurely; mice 7a and 7c were euthanized on day 7. Tissues of mice 5a and 7b were histologically examined, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040380#pone-0040380-g003" target="_blank">figures 3</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040380#pone-0040380-g004" target="_blank">4</a>.</p