Abstract
Porcine reproductive and respiratory syn- drome (PRRS) is considered to be one of the most important infectious diseases impacting the swine industry and is characterized by reproductive failure in late term gestation in sows and respiratory disease in pigs of all ages. The nonstructural protein 9 gene, Nsp9, encoding the RNA-dependent RNA polymerase, is generally regarded as fairly conserved when compared to other viral proteins. Antibodies againstNsp9 will be of great importance for the diagnosis and treatment of the causal agent, PRRS virus. A study was undertaken to generate polyclonal antibodies against the immunodominant Nsp9. For this purpose, the Nsp9 was expressed in Escherichia coli and subsequently used as an antigen to immunize New Zealand rabbits. Antiserum was identified via an indirect ELISA, and then verified based on the ability to react with both naturally and artificially expressed Nsp9. Results of virus neutrali- zation test showed that this antiserum could not neutralize the PRRSV. Nevertheless, this antiserum as a diagnostic core reagent should prove invaluable for further investiga- tions into the mechanism of PRRS pathogenesis.
Highlights
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important threats to the swine industry
The results showed that the recombinant Nsp9 that was expressed in the E. coli exhibits a polyclonal antibody specificity binding reaction (Fig. 6)
To determine whether antiserum could be used for the immunofluorescent detection of the Nsp9 in virus-infected Marc-145 cells, monolayers of Marc-145 cells were infected with PRRS virus (PRRSV)-XH-GD at a MOI of 0.1 at 48 h.p.i
Summary
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important threats to the swine industry. Since it was first identified in the USA in 1987, in Europe in 1990[1] and later in China in 1995, the clinical manifestations of PRRS are severe reproductive failure in sows, which includes early farrowing with stillborn piglets and late term abortion, respiratory distress in piglets and growing pigs. ORF1a and ORF1b comprise 80% of the genome in terms of size and encode the viral replicase polyproteins. ORF2a, ORF2b and ORF 3–7 encode viral structural proteins GP2, E, GP3, GP4, GP5, GP5a, M and N
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