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Classical swine fever (CSF) is a serious infectious disease of pigs which is widespread in Europe. Classical swine fever virus (CSFV) belongs to the Pestivirus genus of the Flaviviridae family. The aim of this work was the application of reverse transcription and polymerase chain reaction (RT-PCR) for the differentiation of pestiviral infections of swine. Sixteen strains of BVDV and 59 strains of CSFV were used in the study. Total RNA was extracted by the Chomczynski method. After reverse transcription of viral RNA, cDNA was amplified using two sets of PCR primers. One set was constructed to detect all pestiviruses, by selecting primers from the highly conservative 5’ untranslated region (5’UTR) of the pestivirus genome. The other set was constructed to detect CSFV strains only, by selecting primers from the E2 coding region. All the pestivirus strains used were detected by PCR with primers from 5’UTR. Only CSFV strains were detected by PCR with primers from E2. The sensitivity of the assay was from 0.1 to 10 TCID₅₀.
Viruses of the genus Pestivirus, family Flaviviridae, are believed to be among the main factors causing economic losses in cattle and pig breeding. Their genomes, consisting of single-stranded RNA with positive polarity and a length of approximately 12.3 kb, have one open reading frame that encodes from 11 to 12 proteins. The virion is surrounded by a lipid membrane. According to the official classification, the genus Pestivirus includes four species: bovine viral diarrhea virus-1 (BVDV-1), bovine viral diarrhea virus-2 (BVDV-2), classical swine fever virus (CSFV) and border disease virus (BDV). To date, several genetically related viruses have been identified, but not yet included in the official classification. They include: giraffe pestivirus isolated from an animal with symptoms of mucosal disease and from cell cultures originating from Kenya, Pronghorn virus from an antelope, HoBi-like viruses detected for the first time in fetal bovine serum and Bungowannah virus causing losses in the Australian domestic pig population. It is suspected that new strains detected in Turkish goat and sheep herds and in Tunisian sheep pox vaccines also belong to pestiviruses. Next-generation sequencing has made it possible to identify another atypical pestivirus of pigs, as well as to discover strains infecting other animals beyond the order of Artiodactyla, such as rats or bats. New emerging strains may pose a threat to the livestock industry.
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