A new method for the diagnosis of pestivirus infections in swine

Researchers of the CReSA and the Centro Nacional de Sanidad Agropecuaria in Cuba (CENSA) have developed a new multiplex RT-PCR method that provides a rapid, highly sensitive, and cost-effective laboratory diagnosis for classical swine fever and other pestivirus infections in swine.

Classical swine fever (CSF), a highly contagious viral infection of domestic pig and wild boar, is one of the most devastating porcine diseases worldwide. The disease is endemic in Asia and prevails in many Central and South American countries, as well as in Eastern Europe. The classical swine fever virus (CSFV) together with the viruses of bovine viral diarrhea and Border disease belongs to the genus Pestivirus within the family Flaviviridae.

Infections in swine with pestiviruses other than CSFV can give rise to a clinical disease that is indistinguishable from CSF. The rapid and accurate differentiation of CSFV from non-CSFV pestiviruses is decisive because whereas CSFV infection of domestic pigs demands strict control measures to prevent vast economic losses for other pestivirus infections in pigs, no immediate zoo-sanitary measures are prescribed. Since different pestiviruses are closely related, both immunologically and genetically, the ruminant's pestivirus infections in swine can result into a false CSF diagnosis.

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 CSFV. The assay proved to be specific when different pestivirus strains from swine and ruminants were evaluated.

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.

Multiplex RT-PCR and single RT-PCR sensitivity assays. (A) Multiplex RT-PCR sensitivity assay for CSFV and panpestivirus detection. (B) RT-PCR sensitivity assay for panpestivirus detection. (C) RT-PCR sensitivity assay for CSFV detection.

The multiplex RT-PCR assay described here provides a sensitive tool for simultaneous detection and rapid differentiation between CSFV and other pestiviruses infections in swine. The reaction can be performed in a rapid and cost-effective manner. This method could be a good alternative for diagnostic laboratories with limited economic resources probably located in developing nations where CSF outbreaks occur frequently.

This study will be published next in the journal Veterinary Microbiology as “A multiplex RT-PCR assay for the rapid and differential diagnosis of classical swine fever and other pestivirus infections”. Arce HD, Pérez LJ, Frías MT, Rosell R, Tarradas J, Núñez JI, Ganges L. 1: Vet Microbiol. 2009 Jun 11. [Epub ahead of print].

To download the full paper:

To contact with the principal investigator of this study at the CReSA:

Dr. Llilianne Ganges Espinosa
Researcher  
Viral Diseases Unit (CReSA)
Edifici CReSA. Campus UAB
08193 Bellaterra (Barcelona) España
Email: llilianne.ganges@cresa.uab.cat
Telephone no.: +34 93.581.46.20
Fax: +34 93 581 44 90


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