Evaluation of a fluorogenic real-time reverse transcription-polymerase chain reaction method for the specific detection of all known serotypes of porcine teschoviruses

J Virol Methods. 2011 Sep;176(1-2):131-4. doi: 10.1016/j.jviromet.2011.05.035. Epub 2011 Jun 1.

Abstract

Performance of a real-time reverse-transcription polymerase chain reaction method for the rapid, simple and reliable detection of porcine teschovirus (PTV) was assessed. The method was based on the use of a set of oligonucleotides consisting of two specific primers and a fluorogenic TaqMan-MGB probe. Reverse transcription and PCR reactions were performed sequentially in one step. As a result the whole procedure was simple and rapid, taking less than 3h for completion. The method reacted in a dose-dependent manner with prototype strains for the eleven known PTV serotypes (PTV1-11), with higher analytical sensitivity than other gel-based RT-PCR methods described, which were performed in parallel to allow for a comparison. The assay did not cross-react with other related viruses or porcine viruses tested. The diagnostic performance of the method was analyzed using a panel of field samples consisting of pig fecal and pig slurry samples. As a conclusion, this technique is adequate and convenient for porcine teschovirus detection, both for diagnosis as well as in environmental investigations.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers
  • Picornaviridae Infections / epidemiology
  • Picornaviridae Infections / veterinary*
  • Picornaviridae Infections / virology
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Serotyping
  • Swine
  • Swine Diseases / epidemiology
  • Swine Diseases / virology*
  • Teschovirus / classification
  • Teschovirus / genetics
  • Teschovirus / isolation & purification*
  • Water Microbiology*
  • Water Pollution*

Substances

  • DNA Primers
  • RNA, Viral