Canal C W, Hotzel I, de Almeida L L, Roehe P M, Masuda A
Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul. Av. Bento Gonçalves, Porto Alegre, RS, Brazil.
Vet Microbiol. 1996 Feb;48(3-4):373-9. doi: 10.1016/0378-1135(95)00156-5.
A reverse transcriptase-polymerase chain reaction (RT-PCR) assay was designed to allow the differentiation of pestiviruses by the expected size of the amplified fragments. One oligonucleotide primer, conserved amongst pestiviruses, and two others specific for either classical swine fever virus (CSFV) or bovine viral diarrhea virus (BVDV), were designed from the 5' non-coding region of the genome. CSFV infected cultures (10 strains) amplified a fragment of an expected size of 200 bp; BVDV cultures (23 strains) or border disease virus (BDV) (2 strains) amplified a fragment of an expected size of 260 bp. The specificity of the amplified fragments was confirmed by restriction enzyme analysis. The threshold of sensitivity was 100 TCID50 for CSFV and 1 TCID50 for BVDV. The RT-PCR described here provides a rapid and sensitive diagnostic tool for the detection and differentiation of CSFV from ruminant pestiviruses.
设计了一种逆转录酶-聚合酶链反应（RT-PCR）检测方法，以通过扩增片段的预期大小来区分瘟病毒。从基因组的5'非编码区设计了一种在瘟病毒中保守的寡核苷酸引物，以及另外两种分别针对经典猪瘟病毒（CSFV）或牛病毒性腹泻病毒（BVDV）的特异性引物。CSFV感染的培养物（10个毒株）扩增出预期大小为200 bp的片段；BVDV培养物（23个毒株）或边界病病毒（BDV）（2个毒株）扩增出预期大小为260 bp的片段。通过限制性酶切分析证实了扩增片段的特异性。CSFV的敏感性阈值为100 TCID50，BVDV为1 TCID50。本文所述的RT-PCR为从反刍动物瘟病毒中检测和区分CSFV提供了一种快速且灵敏的诊断工具。
