The detection of proviral DNA by semi-nested polymerase chain reaction and phylogenetic analysis of Czech Maedi-Visna isolates based on gag gene sequences.
A semi-nested polymerase chain reaction (snPCR) for detecting proviral DNA of ovine lentivirus (OvLV) in peripheral blood mononuclear cells was developed. Primers for snPCR were situated within the gag gene of the Maedi-Visna virus (MVV) genome. A comparison between the snPCR and serological tests (agar gel immunodiffusion test, immunoblot) were performed using 98 ovine blood samples. Thirty (30.6%) of the 98 sheep examined had antibodies specific for the MVV. PCR showed 21 of them to be positive and nine seropositive animals to be PCR negative. Six of the 68 serologically negative sheep were found to be PCR positive, probably due to delayed seroconversion. The PCR amplification products of these six sheep were sequenced and subjected to phylogenetic analysis. The resulting phylogenetic tree of partial gag gene sequences confirmed that the ovine lentivirus genotype in the Czech Republic is more closely related to the prototype MVV isolates than to the caprine arthritis encephalitis viruses.
Celer
V
V
Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Veterinary and Pharmaceutical University, Brno, Czech Republic. celer@dior.ics.muni.cz
Celer
V
V
Nejedlá
E
E
Bertoni
G
G
Peterhans
E
E
Zanoni
R G
RG
D000818:Animals; D001483:Base Sequence; D018153:Czech Republic; D017931:DNA Primers; D004279:DNA, Viral / Q000302:isolation & purification*; D015683:Gene Products, gag / Q000737:chemistry / Q000235:genetics*; D016679:Genome, Viral; D015151:Immunoblotting / Q000662:veterinary; D005779:Immunodiffusion / Q000662:veterinary; D008969:Molecular Sequence Data; D010802:Phylogeny; D011021:Pneumonia, Progressive Interstitial, of Sheep / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011533:Proviruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary; D012756:Sheep; D014790:Visna-maedi virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D004279:DNA, Viral; D015683:Gene Products, gag