Comparison and phylogenetic analysis based on the B2L gene of orf virus from goats and sheep in China during 2009-2011

As a zoonotic infectious disease, orf outbreaks have been reported in China in recent years. However, molecular epidemiology analysis has not been performed for Chinese orf virus (ORFV) strains. Here, we have identified 13 ORFVs from goats and sheep in China between 2009 and 2011. Thirty-four complete B2L sequences were used to construct a phylogenetic tree to elucidate the molecular epidemiology of ORFV in China. Nucleotide sequences of B2L genes of clinical samples and attenuated vaccine strains were aligned and compared. Three genotypes were found by molecular epidemiology analysis. Amino acid substitutions were dispersed among B2 polypeptides from wild and attenuated ORFV strains. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00705-013-1946-6) contains supplementary material, which is available to authorized users

The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus

<p>Abstract</p> <p>Background</p> <p>Prompt detection of PRRSV in the field samples is important for effective PRRS control, thereby reducing the potentially serious economic damage which can result from an outbreak. In this study, a rapid SYBR-based, one step real-time RT-PCR quantitative reverse transcription PCR (qRT-PCR) has been developed for the detection of porcine reproductive and respiratory syndrome virus (PRRSV). Primers were designed based on the sequence of highly conservative region of PRRSV N gene.</p> <p>Results</p> <p>The sensitivity of the real-time qRT-PCR assay was achieved through PRRSV ch-1a RNA for the generation of a standard curve. The detection limit of the assay was found to be 9.6 RNA copies per reaction mixture. This assay had excellent intra- and inter-assay reproducibility as in total 65 field samples were screened for the presence of PRRSV by conventional RT-PCR in parallel with qRT-PCR, and the detection rate increased from 60.0% to 76.9%. Moreover, the specificity result indicated that this assay could reliably differentiate PRRSV from the other swine viral diseases, such as classical swine fever virus (CSFV), swine vesicular disease virus (SVDV) and vesicular exanthema of swine virus (VESV).</p> <p>Conclusion</p> <p>The real-time qRT-PCR assay described in this report allows the rapid, specific and sensitive laboratory detection of PRRSV in field samples.</p

Serodiagnosis of sheeppox and goatpox using an indirect ELISA based on synthetic peptide targeting for the major antigen P32

<p>Abstract</p> <p>Background</p> <p>Sheeppoxvirus (SPPV), goatpoxvirus (GTPV) and lumpy skin disease virus (LSDV) of cattle belong to the <it>Capripoxvirus </it>genus of the <it>Poxviridae </it>family and can cause significant economic losses in countries where they are endemic. Despite the considerable threat that these viruses pose to livestock production and global trade in sheep, goats, cattle and their products, convenient and effective serodiagnostic tools are not readily available. Toward this goal, two synthetic peptides corresponding to the major antigen P32 were synthesized. These synthetic peptides were then used as antigen to develop an ELISA method to detect anti-SPPV and GTPV antibodies.</p> <p>Results</p> <p>The results indicated that the optimal concentration of coated recombinant antigen was 0.2 μg per well for a serum dilution of 1:10. The ELISA performed favorably when sera from sheep immunized experimentally were tested.</p> <p>Conclusion</p> <p>This assay offers the prospect of synthetic peptide as antigens for indirect ELISA to detect SPPV and GTPV antibody in sheep and goat sera.</p

Analysis of pig serum proteins based on shotgun liquid chromatography-tandem mass spectrometry

Recent advances in proteomics technologies have opened up significant opportunities for future applications. We used shotgun liquid chromatography, coupled with tandem mass spectrometry (LC-MS/MS) to determine the proteome profile of healthy pig serum. Samples of venous blood were collected and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation and in-gel trypsin digestion. The peptides were then processed using shotgun LC-MS/MS. Serum proteins were subjected to protein identification and bioinformatics analysis. A total of 392 proteins were identified, and 179 were annotated according to their molecular functions and biological processes, excluding 142 hypothetical proteins and 71 immune globulins. To the best of our knowledge, this represents the first porcine serum proteomics analysis based on shotgun LC-MS/MS. This method and the resulting proteomics information may prove valuable for ensuring good animal welfare practice and for monitoring swine health and disease status.Keywords: Analysis, pig serum, shotgun coupled with tandem mass spectrometry (LC-MS/MS

Diagnosis and phylogenetic analysis of Orf virus from goats in China: a case report

<p>Abstract</p> <p>Background</p> <p>Orf virus (ORFV) is the etiological agent of contagious pustular dermatitis and is the prototype of the genus Parapoxvirus (PPV). It causes a severe exanthematous dermatitis that afflicts domestic and wild small ruminants.</p> <p>Case presentation</p> <p>In the present study, an outbreak of proliferative dermatitis in farmed goats. The presence of ORFV in tissue scrapings from the lips was confirmed by B2L gene polymerase chain reaction (PCR) amplification. The molecular characterization of the ORFV was performed using PCR amplification, DNA sequencing and phylogenetic analysis of the B2L gene.</p> <p>Conclusion</p> <p>The results of this investigation indicated that the outbreak was caused by infection with an ORFV that was closely related genetically to Nantou (DQ934351), which was isolated from the Tai wan province of China and Hoping (EU935106), which originated from South Korea in 2008. This is the first report of the phylogenetic analysis of ORFV from goats in China.</p