Sequences of primers and probes.

<p>FAM 5′ modification was used for IL-1β, IL-2, IL-4, IL-6, IL-8, IFN-α, TNF-α; HEX for β-Actin; Texas Red for GAPDH. BHQ (Black-Hole-Quencher)-1 was used for 3′ modifications of IL-1β, IL-2, IL-4, IL-6, IFN-α, β-Actin and GAPDH; BHQ-2 was used for of TNF-α and IL-8. Corresponding references are given in the right column. Sequences marked with “this study” were created by the use of “Primer-BLAST” available on NCBI GenBank.</p><p>F =  Forward Primer; R =  Reverse Primer; P =  Probe; bp = base pairs.</p><p>Sequences of primers and probes.</p

In-vitro cytokine stimulation.

<p>The stimulating agents are presented along with the corresponding target cytokines as well as background information about their functionality.</p><p>LPS = <i>Salmonella typhimurium</i> lipopolysaccharid; a component of the outer gram positive bacteria membrane, antigenic effect on PBMCs.</p><p>PGN =  <i>Staphylococcus aureus</i> peptidoglycan; a stabilizing macro molecule in the cell wall of gram positive bacteria; antigenic effect on PBMCs.</p><p>ConA =  Concanavalin A; a lectin from the jack bean, mitogenic effect (especially on T-cells).</p><p>PWM = Pokeweed mitogen; a lectin of the American pokeweed, activating effect on B- and T-cells.</p><p>PHA =  Phytohemagglutinin; a herbal lectin, mitogenic effect (especially on T-cells).</p><p>In-vitro cytokine stimulation.</p

Composition of the synthetic standard gene comprising all target cytokines (IL-2, IL-4, IL-8, TNF-α, IFN-α, IL-6, IL-1β) and internal reference genes (β-Actin, GAPDH, HPRT (Hypoxanthin-Guanin-Phosphoribosyltransferase), starting with the T7-promotor sequence and concluding with the NOD1 restriction site as initial point for linearization and transformation to RNA.

<p>Each target cytokine was included with a nucleotide overhang of approximately 50 base pairs (bp) prior to forward primer sequence. In total, the synthetic standard gene comprises 1464 bp.</p

Detection of normalized TNF-α gene expression (ΔΔCq) in leukocyte samples from CSFV infected pigs.

<p>Samples were obtained from two different pig breeds at days 0, 2, 4, and 7 post infection (dpi). Results are given as mean values: in total from all animals (TNF-α overall mean) and separately for each breed (Mean Breed 1, Mean Breed 2). Bars indicate standard deviations.</p

Samples from different animal trials (n = 402) used for assay validation.

<p>Samples were chosen to represent different pig species (wild boar, domestic pigs) and inoculation status (CSFV infection/vaccination, ASFV infection, corresponding control animals). Moreover, PBMC and EDTA blood samples were included.</p><p>Samples from different animal trials (n = 402) used for assay validation.</p

Detection of normalized IL-8 gene expression (ΔΔC_q) in leukocyte samples from CSFV infected pigs.

<p>Samples were obtained from two different pig breeds at days 0, 2, 4, and 7 post infection (dpi). Results are given as mean values: in total from all animals (IL-8 overall mean) and separately for each breed (Mean Breed 1, Mean Breed 2). Bars indicate standard deviations.</p

Comparative amplifications of 10-fold diluted standard RNA ranging from 2×10² to 2×10⁷ copies/µl of single target and multiplex assays of all target cytokines (FAM).

<p>The vertical axis demonstrate fluorescence levels (relative fluorescence units, RFU), the horizontal axis shows the number of cycles. Comparative illustrations of end fluorescence levels of standard curves are shown illustrating higher end fluorescences in single target compared triplex assays in which target cytokines were detected simultaneously with β-Actin (HEX) and GAPDH (TR).</p

Additional file 1: of Engineered recombinant protein products of the avian paramyxovirus type-1 nucleocapsid and phosphoprotein genes for serological diagnosis

Cut-off values of indirect ELISAs based on recombinant avian paramyxovirus NP and P gene products. Provides data that elucidate the fixing of a threshold for indirect ELISA (DOCX 12 kb

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

Additional file 1: of Influence of cell type and cell culture media on the propagation of foot-and-mouth disease virus with regard to vaccine quality

Table S1.1. Nucleotide changes in the VP1 coding region of FMDV type O1 Manisa during serial passaging in adherent BHK21 cells. Table S1.2. Nucleotide changes in the VP1 coding region of FMDV type O1 Manisa during serial passaging in BHK-2P suspension cells. Table S1.3. Nucleotide changes in the capsid-coding region of FMDV type A24 Cruzeiro during serial passaging in adherent BHK21 cells. Table S1.4. Nucleotide changes in the capsid-coding region of FMDV type A24 Cruzeiro during serial passaging in BHK-2P suspension cells. (DOCX 17 kb