A multiplex RT-PCR assay for the rapid and differential diagnosis of classical swine fever and other pestivirus infections

Heidy Díaz de Arce, Lester J. Pérez, Maria T. Frías, Rosa Rosell, Joan Tarradas, José I. Núñez, Llilianne Ganges

Research output: Contribution to journalArticleResearchpeer-review

24 Citations (Scopus)

Abstract

Classical swine fever is a highly contagious viral disease causing severe economic losses in pig production almost worldwide. All pestivirus species can infect pigs, therefore accurate and rapid pestivirus detection and differentiation is of great importance to assure control measures in swine farming. Here we describe the development and evaluation of a novel multiplex, highly sensitive and specific RT-PCR for the simultaneous detection and rapid differentiation between CSFV and other pestivirus infections in swine. The universal and differential detection was based on primers designed to amplify a fragment of the 5′ non-coding genome region for the detection of pestiviruses and a fragment of the NS5B gene for the detection of classical swine fever virus. The assay proved to be specific when different pestivirus strains from swine and ruminants were evaluated. The analytical sensitivity was estimated to be as little as 0.89 TCID50. The assay analysis of 30 tissue homogenate samples from naturally infected and non-CSF infected animals and 40 standard serum samples evaluated as part of two European Inter-laboratory Comparison Tests conducted by the European Community Reference Laboratory, Hanover, Germany proved that the multiplex RT-PCR method provides a rapid, highly sensitive, and cost-effective laboratory diagnosis for classical swine fever and other pestivirus infections in swine. © 2009 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)245-252
JournalVeterinary Microbiology
Volume139
Issue number3-4
DOIs
Publication statusPublished - 18 Nov 2009

Keywords

  • Classical swine fever virus
  • Differential diagnosis
  • Multiplex RT-PCR
  • Pestivirus

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