A novel RNA-based in situ hybridization to detect Seneca Valley virus in neonatal piglets and sows affected with vesicular disease

PLoS One. 2017 Apr 10;12(4):e0173190. doi: 10.1371/journal.pone.0173190. eCollection 2017.

Abstract

Seneca Valley virus (SVV) is the causative agent of an emerging vesicular disease in swine, which is clinically indistinguishable from other vesicular diseases such as foot-and-mouth disease. In addition, SVV has been associated with neonatal mortality in piglets. While a commercial SVV qRT-PCR is available, commercial antibodies are lacking to diagnose SVV infections by immunohistochemistry (IHC). Thus, a novel in situ hybridization technique-RNAscope (ISH) was developed to detect SVVRNA in infected tissues. From a total of 78 samples evaluated, 30 were positive by qRT-PCR and ISH-RNA, including vesicular lesions of affected sows, ulcerative lesions in the tongue of piglets and various other tissues with no evidence of histological lesions. Nineteen samples were negative for SVV by qRT-PCR and ISH-RNA. The Ct values of the qRT-PCR from ISH-RNA positive tissues varied from 12.0 to 32.6 (5.12 x 106 to 5.31 RNA copies/g, respectively). The ISH-RNA technique is an important tool in diagnosing and investigating the pathogenesis of SVV and other emerging pathogens.

MeSH terms

  • Animals
  • Animals, Newborn
  • Female
  • Heart / virology
  • Immunohistochemistry
  • In Situ Hybridization
  • Lymph Nodes / metabolism
  • Lymph Nodes / pathology
  • Lymph Nodes / virology
  • Myocardium / metabolism
  • Myocardium / pathology
  • Necrosis / metabolism
  • Necrosis / pathology
  • Necrosis / virology
  • Paranasal Sinuses / metabolism
  • Paranasal Sinuses / pathology
  • Paranasal Sinuses / virology
  • Picornaviridae Infections / diagnosis
  • Picornaviridae Infections / metabolism
  • Picornaviridae Infections / pathology
  • Picornaviridae Infections / veterinary*
  • Picornaviridae*
  • RNA, Viral / metabolism
  • Real-Time Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Spleen / metabolism
  • Spleen / pathology
  • Spleen / virology
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / metabolism
  • Swine Diseases / pathology
  • Swine Diseases / virology*
  • Tongue / metabolism
  • Tongue / pathology
  • Tongue / virology

Substances

  • RNA, Viral

Grants and funding

TPR was supported by CAPES - Brazil. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.