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Diagnostyka roznicowa pestiwirusowych zakazen swin przy uzyciu amplifikacji DNA in vitro

100%

Stadejek T. , Pejsak Z.

nr 07

466-469

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.