RESUMO
Sixty-seven serum samples and 43 pathological material samples from wild boars, taken in 5 regions of Russia and in the Kharkov Region of the Ukraine in 2002 to 2005 were studied. Wild boars in some regions of Russia were shown to be carriers of Aujeszky's disease virus, porcine parvovirus, porcine circovirus type 2, lymphotropic herpesvirus-1, porcine cytomegalovirus, Mycoplasma hyopneumoniae and Pasteurella multocida. The classical swine fever (CSF) virus genome was detected by polymerase chain reaction in the samples from 10 wild boars from 2 Russian regions (the Tver and Moscow Regions). Sequencing of the E2 gene 5'-terminal region of detected CSF virus isolates showed that they were closely related to two field virus isolates early found in domestic pigs in the Moscow and Vladimir Regions, which suggests that there is an epizootic relation between the SCF outbreaks among wild boars and domestic pigs in these regions. Tests for porcine reproductive and respiratory syndrome, transmissible porcine gastroenteritis, porcine influenza, enteroviruses, and actinobacillus-induced pleuropneumonia were negative in all the regions under study.
Assuntos
Doenças Transmissíveis/veterinária , Reservatórios de Doenças/veterinária , Monitoramento Ambiental , Sus scrofa/microbiologia , Animais , Animais Domésticos/sangue , Animais Domésticos/virologia , Anticorpos Antibacterianos/sangue , Anticorpos Antivirais/sangue , Peste Suína Clássica/epidemiologia , Vírus da Febre Suína Clássica/genética , Vírus da Febre Suína Clássica/imunologia , Vírus da Febre Suína Clássica/isolamento & purificação , Doenças Transmissíveis/sangue , Doenças Transmissíveis/epidemiologia , Reservatórios de Doenças/microbiologia , Monitoramento Epidemiológico , Genoma Viral/genética , Mycoplasma hyopneumoniae/imunologia , Infecções por Pasteurella/epidemiologia , Infecções por Pasteurella/veterinária , Pasteurella multocida/imunologia , Pneumonia Suína Micoplasmática/epidemiologia , Reação em Cadeia da Polimerase , Federação Russa/epidemiologia , Estudos Soroepidemiológicos , Suínos/microbiologia , Doenças dos Suínos/sangue , Doenças dos Suínos/epidemiologia , Ucrânia/epidemiologia , Viroses/sangue , Viroses/epidemiologia , Viroses/veterináriaRESUMO
Two approaches for simultaneous identification of both Foot-and-mouth disease virus (FMDV) and Swine vesicular disease virus (SVDV) are described: (1) a single-step reverse transcription-PCR with three primers and (2) a PCR-ELISA assay with two universal primers for genome amplification and two virus-specific probes for identification. These methods are based on the use of 3D gene universal PCR primers, the structure of which was optimized and refined due to the close relationship between the two viruses belonging to different genera of the Picornaviridae family. In procedure (1), a three-primer PCR containing one universal antisense primer and two virus-specific primers was shown to differentiate between FMDV and SVDV in one reaction, due to the different length of the amplified DNA fragments (600 and 340 base pairs, respectively). In procedure (2), the two viruses were identified by PCR-ELISA, i.e. PCR for the 3D gene followed by two parallel hybridizations with FMDV and SVDV-specific probes in microplate wells and ELISA detection. The application of universal primers could halve the number of PCR experiments in both cases, as compared to the usual virus-specific PCR procedures. Also, we investigated the 3D gene structure of several SVDV strains isolated at different times. No essential changes were detected in the regions coding for conserved motifs of the RNA-dependent RNA polymerase recognized by our universal primers. The multi-primer PCR was successfully tested on 38 FMDV and 15 SVDV strains, and the PCR-ELISA on 32 FMDV and 16 SVDV strains including clinical material from disease cases.
Assuntos
Antígenos Virais/genética , RNA Polimerases Dirigidas por DNA/genética , Enterovirus Humano B/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/métodos , Vírus da Febre Aftosa/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Proteínas não Estruturais Virais/genética , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Elementos Antissenso (Genética) , Sequência de Bases , Primers do DNA/síntese química , Enterovirus Humano B/genética , Vírus da Febre Aftosa/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Alinhamento de SequênciaRESUMO
A possibility of using the amplification of gene HN fragment in combination with nucleotide cDNA sequencing for the purpose of identification and strain differentiation of bovine parainfluenza-3 virus was demonstrated. A comparative analysis of the primary structure in the studied HN gene fragment revealed 2 genetic groups among the investigated virus' strains and isolates. Group 1 is made up of Northern American viral strains and of Russian isolates, whose primary structure has a high level of homology to the primary SF-4/32 strain structure; group 2 comprises the virus' Russian isolates with a high level of homology to the mentioned strains to Japanese strains' sequences. The biggest differences between the studied strains and the viral isolates amounted to around 8%, when the nucleotide sequences were compared, and to around 4%, when the corresponding amino-acid sequences were compared.
