To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rab...To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.展开更多
With the purpose to determine the frequency and type of cardiac lesions in naturally exposed dogs to Trypanosoma cruzi, ninety one stray dogs, capture by the Canine and Feline Control Center （dog pound） from the mun...With the purpose to determine the frequency and type of cardiac lesions in naturally exposed dogs to Trypanosoma cruzi, ninety one stray dogs, capture by the Canine and Feline Control Center （dog pound） from the municipality of Merida, were studied. Before euthanasia, blood samples were taken to detect 72 cruzi antigens by indirect immunofluorescence antibody test and Western Blot and to detect the genome of parasite by Polymerase Chain Reaction. Immediately after euthanasia, hearts were macroscopically evaluated and a sample of the middle right atrial wall of each dog was taken for histopathological analyses. DNA was also obtained from paraffin blocks of seropositives animals with microscopic lesions to detect 72 cruzi genome. Of ninety one dogs, thirteen were seropositive. All seropositive dogs showed an association （P 〈 0.05） with lymphocytoplasmatic myocarditis. The presence of the 72 cruzi genome was also detected by PCR in cardiac septum tissue of seropositive dogs and in all the cases with microscopic lesions indicating the high pathogenicity of the local circulating strain. No association with macroscopic lesions was observed in seropositive dogs. Also, the presence of Dirofilaria immitis （D. immitis） was found in 6% of dogs evaluated. This study demonstrates a high tropism to cardiac tissue and virulence of the strains of 72 cruzi circulating in the studied dog population.展开更多
基金National Department Public Benefit Research Foundation (201103032)Pathogens Network Monitoring Technology Research (2008ZX10004-008)
文摘To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.
文摘With the purpose to determine the frequency and type of cardiac lesions in naturally exposed dogs to Trypanosoma cruzi, ninety one stray dogs, capture by the Canine and Feline Control Center （dog pound） from the municipality of Merida, were studied. Before euthanasia, blood samples were taken to detect 72 cruzi antigens by indirect immunofluorescence antibody test and Western Blot and to detect the genome of parasite by Polymerase Chain Reaction. Immediately after euthanasia, hearts were macroscopically evaluated and a sample of the middle right atrial wall of each dog was taken for histopathological analyses. DNA was also obtained from paraffin blocks of seropositives animals with microscopic lesions to detect 72 cruzi genome. Of ninety one dogs, thirteen were seropositive. All seropositive dogs showed an association （P 〈 0.05） with lymphocytoplasmatic myocarditis. The presence of the 72 cruzi genome was also detected by PCR in cardiac septum tissue of seropositive dogs and in all the cases with microscopic lesions indicating the high pathogenicity of the local circulating strain. No association with macroscopic lesions was observed in seropositive dogs. Also, the presence of Dirofilaria immitis （D. immitis） was found in 6% of dogs evaluated. This study demonstrates a high tropism to cardiac tissue and virulence of the strains of 72 cruzi circulating in the studied dog population.
