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1.
Genet Mol Res ; 14(4): 14530-8, 2015 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-26600512

RESUMO

We conducted a phylogenetic analysis of 22 strains of bovine leukemia virus obtained by polymerase chain reaction to amplify a 582-base pair fragment of the transcriptional regulatory region 5' long terminal repeat (LTR). Twenty-two samples of proviral DNA from peripheral blood mononuclear cells containing bovine leukemia virus from naturally infected bovine from 4 distinct geographic regions in Brazil were investigated. The products obtained by polymerase chain reaction were subjected to direct sequencing and sequence alignment. Fragments of 422 nucleotides were obtained, located between positions -118 and +303 base pairs of the 5'LTR. These fragments corresponded to 80% of the LTR region and included 56% of sub-region U3, 100% of R, and 82.5% of U5. Phylogenetic analysis of these sequences showed a high conservation degree in the 5'LTR region, with 5 well defined groups. However, a hotspot occurrence in the R-U5 region was also observed, which contained 40% of all nucleotide variability observed.


Assuntos
Variação Genética , Vírus da Leucemia Bovina/genética , Filogenia , Sequências Repetidas Terminais/genética , Animais , Sequência de Bases , Brasil , Bovinos , DNA Viral/genética
2.
Genet Mol Res ; 13(1): 1717-23, 2014 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-24535910

RESUMO

This article reports the selection of bovine leukemia virus (BLV) variants after continuous passage in cell lines or experimental animals. Two wild BLV strains isolated from 2 naturally infected Holstein dairy cows in Brazil (cow codes: 485 and 141) were used for the experimental infection of 1 sheep and FLK cells, and 1 rabbit and CC81 cells. Viral DNA was isolated several months after infection, and env gene nucleotide and amino acid sequences of the "passaged" variants were compared against the 2 original infecting wild strains. The sequences of the original infecting wild strains were not recovered after their replication in the cell lines or experimental animals. These results indicate that genetic variation occurred after BLV replication in vivo and in vitro, with new variants being selected.


Assuntos
DNA Viral/genética , Genes env , Vírus da Leucemia Bovina/genética , Replicação Viral/genética , Animais , Sequência de Bases , Brasil , Bovinos , Divisão Celular , Linhagem Celular , Vírus da Leucemia Bovina/patogenicidade , Coelhos , Ovinos
3.
Genet Mol Res ; 11(3): 2182-99, 2012 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-22653674

RESUMO

Equine infectious anemia caused by equine infectious anemia virus is an important disease due to its high severity and incidence in animals. We used a phage display library to isolate peptides that can be considered potential markers for equine infectious anemia diagnosis. We selected peptides using IgG purified from a pool comprised of 20 sera from animals naturally infected with equine infectious anemia virus. The diagnostic potential of these peptides was investigated by ELISA, Western blot and dot blot with purified IgG and serum samples. Based on the results, we chose a peptide mimetic for glycoprotein gp45 epitopes of equine infectious anemia virus, with potential for use as an antigen in indirect diagnostic assays. Synthesis of this peptide has possible applications for the development of new diagnostic tools for this disease.


Assuntos
Anemia Infecciosa Equina/sangue , Anemia Infecciosa Equina/diagnóstico , Cavalos/sangue , Cavalos/virologia , Peptídeos , Sequência de Aminoácidos , Animais , Western Blotting , Biologia Computacional , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/imunologia , Vírus da Anemia Infecciosa Equina/isolamento & purificação , Dados de Sequência Molecular , Biblioteca de Peptídeos , Peptídeos/química
4.
Braz J Microbiol ; 43(4): 1632-40, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24031995

RESUMO

Suid herpesvirus 1 (SuHV-1) is the causative agent of pseudorabies (PR), a disease of great importance due to the huge losses it causes in the swine industry. The aim of this study was to determine a method for genotyping SuHV-1 based on partial sequences of the gene coding for glycoprotein C (gC) and to elucidate the possible reasons for the variability of this region. A total of 109 gCsequences collected from GenBank were divided into five major groups after reconstruction of a phylogenetic tree by Bayesian inference. The analysis showed that a portion of gC (approximately 671 bp) is under selective pressure at various points that coincide with regions of protein disorder. It was also possible to divide SuHV-1 into five genotypes that evolved under different selective pressures. These genotypes are not specific to countries or continents, perhaps due to multiple introduction events related to the importation of swine.

5.
Infect Genet Evol ; 103: 105322, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35753623

RESUMO

Cross-species transmission events and mixed infection of small ruminant lentiviruses (SRLVs) were studied in seven goats and two sheep from three small ruminant mixed flocks from Northeast and Southeast Brazil. Genetic and antigenic analyses with gag/env genes and ELISA multiepitope SU1/SU5 recombinant antigens were carried out, respectively. The genetic analysis of gag and env sequences showed high viral diversity in both species, MVV-like (subtype A1) and CAEV-like B1 in goats, and CAEV-like (subtype B1) in sheep, revealing SRLV interspecies transmission from sheep to goats and vice versa in Brazilian farms. Two Brazilian caprine lentiviruses were segregated in two new genetic clades based on gag analyses, which suggests a new classification into heterogenic genotype A. Furthermore, goat isolates were grouped into subtype A1 and B1 clusters. Cross-reactive antibodies were detected in goats using ELISA with a recombinant antigen carrying SU1 and SU5 immunodominant epitopes; the results showed anti-CAEV and MVV antibodies in goats and anti-CAEV antibodies in sheep. This result can be associated with the high divergence in the V4 region due to SRLV variability. All results confirm cross-species infection of SRLV in Brazilian mixed herds.


Assuntos
Doenças das Cabras , Infecções por Lentivirus , Doenças dos Ovinos , Animais , Brasil/epidemiologia , Cabras , Lentivirus/genética , Infecções por Lentivirus/veterinária , Filogenia , Ruminantes , Ovinos
6.
Arch Virol ; 155(10): 1687-92, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20607317

RESUMO

The aim of this study was to analyze the disordered regions (DRs) in the envelope and tegument proteins of three closely related herpesviruses: bovine herpesvirus 1, bovine herpesvirus 5 and suid herpesvirus 1. Tegument proteins showed a greater percentage of DRs than the envelope proteins did. Regions of disorder were found in the less conserved portions of the proteins, N-terminal and C-terminal regions, and, in a few cases, functionally important sites. The presence of DRs is an important factor in the evolution of viruses, representing points where positive pressure led to structural changes.


Assuntos
Herpesvirus Bovino 1/genética , Herpesvirus Suídeo 1/genética , Herpesvirus Bovino 5/genética , Proteínas Estruturais Virais/genética , Evolução Molecular , Conformação Proteica , Homologia de Sequência
7.
Genet Mol Res ; 9(3): 1591-8, 2010 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-20730710

RESUMO

We compared three different protocols for DNA extraction from horse peripheral blood mononuclear cells (PBMC) and lung fragments, determining average final DNA concentration, purity, percentage of PCR amplification using beta-actin, and cost. Thirty-four samples from PBMC, and 33 samples from lung fragments were submitted to DNA extraction by three different protocols. Protocol A consisted of a phenol-chloroform and isoamylic alcohol extraction, Protocol B used alkaline extraction with NaOH, and Protocol C used the DNAzol((R)) reagent kit. Protocol A was the best option for DNA extraction from lung fragments, producing high DNA concentrations, with high sensitivity in PCR amplification (100%), followed by Protocols C and B. On the other hand, for PBMC samples, Protocol B gave the highest sensitivity in PCR amplification (100%), followed by Protocols C and A. We conclude that Protocol A should be used for PCR diagnosis from lung fragment samples, while Protocol B should be used for PBMC.


Assuntos
DNA/isolamento & purificação , Leucócitos Mononucleares/metabolismo , Pulmão/metabolismo , Animais , Cavalos , Reação em Cadeia da Polimerase
8.
Genet Mol Res ; 9(1): 266-76, 2010 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-20198582

RESUMO

Epsilon toxin produced by Clostridium perfringens types B and D causes enterotoxemia in sheep, goats and calves. Enterotoxemia can cause acute or superacute disease, with sudden death of the affected animal. It provokes huge economic losses when large numbers of livestock are affected. Therapeutic intervention is challenging, because the disease progresses very rapidly. However, it can be prevented by immunization with specific immunogenic vaccines. We cloned the etx gene, encoding epsilon toxin, into vector pET-11a; recombinant epsilon toxin (rec-epsilon) was expressed in inclusion bodies and was used for animal immunization. Serum protection was evaluated and cross-serum neutralization tests were used to characterize the recombinant toxin. To analyze the potency of the toxin (as an antigen), rabbits were immunized with 50, 100 or 200 microg recombinant toxin, using aluminum hydroxide gel as an adjuvant. Titers of 10, 30 and 40 IU/mL were obtained, respectively. These titers were higher than the minimum level required by the European Pharmacopoeia (5 IU/mL) and by the USA Code of Federal Regulation (2 IU/mL). This rec-epsilon is a good candidate for vaccine production against enterotoxemia caused by epsilon toxin of C. perfringens type D.


Assuntos
Toxinas Bacterianas/genética , Toxinas Bacterianas/imunologia , Imunização , Algoritmos , Animais , Anticorpos Neutralizantes/biossíntese , Clonagem Molecular , Camundongos , Filogenia , Coelhos , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação
9.
Colloids Surf B Biointerfaces ; 177: 377-388, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30785035

RESUMO

It is estimated that over 100 million people have been infected with human immunodeficiency virus (HIV-1) resulting in approximately 30 million deaths globally. Herein, we designed and developed novel nano-immunoconjugates using gold nanoparticles (AuNPs) and carboxymethylcellulose (CMC) biopolymer, which performed simultaneously as an eco-friendly in situ reducing agent and surface stabilizing ligand for the aqueous colloidal process. These AuNPs-CMC nanocolloids were biofunctionalized with the gp41 glycoprotein receptor (AuNPs-CMC-gp41) or HIV monoclonal antibodies (AuNPs-CMC_PolyArg-abHIV) for detection using the laser light scattering immunoassay (LIA). These AuNPs-CMC bioengineered nanoconjugates were extensively characterized by morphological and physicochemical methods, which demonstrated the formation of spherical nanocrystalline colloidal AuNPs with the average size from 12 to 20 nm and surface plasmon resonance peak at 520 nm. Thus, stable nanocolloids were formed with core-shell nanostructures composed of AuNPs and biomacromolecules of CMC-gp41, which were cytocompatible based on in vitro cell viability results. The AuNPs-CMC-gp41 nanoconjugates were tested against HIV monoclonal antibodies conjugates (AuNPs-CMC_PolyArg-abHIV) using the light scattering immunoassay (LIA) where they behaved as active nanoprobes for the detection at nM level of HIV-1 antigenic proteins. This strategy offers a novel nanoplatform for creating bioprobes using green nanotechnology for the detection of HIV-1 and other virus-related diseases.


Assuntos
Carboximetilcelulose Sódica/química , Ouro/química , HIV-1/isolamento & purificação , Imunoensaio , Lasers , Nanopartículas/química , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Sobrevivência Celular , Coloides/química , Ouro/imunologia , Células HEK293 , HIV-1/imunologia , Humanos , Conformação Molecular , Tamanho da Partícula , Propriedades de Superfície
11.
Arq. bras. med. vet. zootec. (Online) ; 71(2): 711-714, mar.-abr. 2019. tab
Artigo em Inglês | VETINDEX, LILACS | ID: biblio-1038589

RESUMO

O vírus da imunodeficiência bovina é o agente causador da imunodeficiência viral bovina que é conhecido por infectar bovinos em todo o mundo. Como em outras infecções por retrovírus, os hospedeiros desenvolvem uma infecção de longo prazo e a maioria dos animais infectados permanece assintomática. O objetivo deste estudo foi detectar DNA proviral BIV em amostras de sangue de bovinos e estimar a ocorrência de infecção no estado de Minas Gerais, Brasil. Amostras de sangue de 391 bovinos foram coletadas de duas regiões do estado, Zona da Mata e Central. O DNA proviral foi detectado por reação em cadeia da polimerase semi-nested (SN-PCR). Os resultados de SN-PCR indicaram uma ocorrência de BIV de 12,5% no estado. Os produtos amplificados foram confirmados como BIV por sequenciamento e a similaridade da sequência de nucleotídeos com a estirpe de referência (R-29) foi de 99%. Este é o primeiro estudo que relata a presença do BIV em Minas Gerais, Brasil. Os resultados indicam a necessidade de realizar um estudo detalhado sobre a prevalência da infecção por BIV no Brasil.(AU)


Assuntos
Animais , Bovinos , Infecções por Lentivirus/sangue , Vírus da Imunodeficiência Bovina/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária
12.
Vet Microbiol ; 157(3-4): 320-32, 2012 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-22310073

RESUMO

The genetically distinct wild horse herds inhabiting Shackleford Banks, North Carolina are probably the direct descendents of Spanish stock abandoned after failed attempts to settle mid-Atlantic coastal regions of North America in the Sixteenth Century. In a 1996 island survey, 41% of the gathered horses were discovered seropositive for Equine Infectious Anemia Virus (EIAV) with additional cases identified in 1997 and 1998. As a result of their unique genetic heritage, EIAV seropositive individuals identified in the two latter surveys were transferred to a quarantine facility on the mainland. In September 2008 two of the horses SB1 and SB2 after 10 and 11 years in quarantine respectively, developed clinical signs of EIA. In the case of SB2 these were so severe that the only humane option was euthanasia. Although SB1, survived it experienced a second clinical episode one month later. In May 2009, a third horse in quarantine, SB3, developed extremely severe clinical EIA and was euthanized. This demonstrates naturally infected long-term inapparent carriers can experience recrudescence of very severe disease many years after initial exposure to EIAV. Phylogenetic analysis of complete EIAV gag gene sequences obtained from each of three Shackleford horses demonstrated they were infected with very closely related viruses. Although these were distinguishable from all other strains examined, they belong to a monophyletic group comprising almost exclusively of New World isolates that is distinct from a number of recently characterized Central European EIAV strains.


Assuntos
Anemia Infecciosa Equina/virologia , Genes gag , Cavalos/virologia , Vírus da Anemia Infecciosa Equina/genética , Filogenia , Sequência de Aminoácidos , Animais , Genes Virais , Vírus da Anemia Infecciosa Equina/classificação , Vírus da Anemia Infecciosa Equina/isolamento & purificação , Dados de Sequência Molecular , North Carolina , RNA Viral/genética , Análise de Sequência de RNA
13.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);66(6): 1695-1702, 12/2014. graf
Artigo em Inglês | LILACS | ID: lil-735770

RESUMO

Brucella ovis is a major cause of epididymitis in sexually mature rams, resulting in subfertility, infertility, and economic losses for the sheep industry worldwide. The aim of this study was to develop an indirect ELISA (iELISA) using recombinant proteins, namely rBoP59 and rBP26, as antigens for serological diagnosis of B. ovis infection. The BoP59 and BP26 recombinant proteins were expressed in E. coli and purified by affinity chromatography. Antigenicity was tested by Western blot and iELISA. Standardization of iELISA was performed with 500ng and 1µg BoP59 and rBP26 per well, testing serum from uninfected and experimentally infected rams. rBP26 was effective in distinguishing positive from negative rams. The rBP26 iELISA developed in this study is the first to use a completely purified rBP26 as antigen resulting in high sensitivity (100%) and specificity (90.2%), and an overall accuracy equal to 1.0...


Brucella ovis é uma das principais causas de epididimite em carneiros sexualmente maduros, resultando em subfertilidade e infertilidade e consequentes perdas econômicas para a ovinocultura em todo o mundo. O objetivo deste estudo foi desenvolver ELISA indireto (ELISAi), utilizando como antígeno proteínas recombinantes BoP59r e BP26r para diagnóstico da infecção por B. ovis. BoP59r e BP26r foram expressas em E. coli e purificadas por cromatografia de afinidade e a antigenicidade testada por Western blot e ELISAi. A padronização do ELISAi foi realizada testando 500 ng e 1 µg de BoP59r e BP26r por poço e soros de carneiros infectados e não infectados. A BP26r foi eficiente em diferenciar ovinos negativos de positivos. O ELISAi com BP26 desenvolvido neste estudo foi o primeiro a utilizar BP26 completamente purificada como antígeno, resultando em elevada sensibilidade (100%) e sensibilidade (90,2%), com acurácia igual a 1,0...


Assuntos
Animais , Antígenos/análise , Brucella ovis , Epididimite/veterinária , Ovinos/imunologia , Ensaio de Imunoadsorção Enzimática , Ensaio de Imunoadsorção Enzimática/veterinária
14.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);65(3): 801-808, June 2013. tab
Artigo em Português | LILACS | ID: lil-679116

RESUMO

O objetivo deste trabalho foi desenvolver uma PCR em tempo real (qPCR) para o diagnóstico rápido e sensível da doença de Aujeszky. Os iniciadores amplificaram um fragmento de 123 pares de base do gene codificante da glicoproteína D. A qPCR foi testada em 25 amostras de cérebro de suíno positivas e 85 amostras negativas para DA no isolamento viral e na soroneutralização. A sensibilidade analítica foi calculada com acréscimo de um isolado brasileiro do SuHV-1 titulado em amostras de cérebro de suíno negativas na soroneutralização e na PCR. A técnica apresentou sensibilidade analítica de 10-0,5 TCID50/50µL. A qPCR foi capaz de distinguir reações inespecíficas devido a dímero de oligonucleotídeos iniciadores ou amplificações, além do alvo designado (evitando, assim, os falso-positivos), e de obter resultados rápidos.


The aim of this study was to validate a low-cost real-time PCR for a quick and sensitive diagnosis of the disease. The fluorofore used was a DNA intercalating agent, one of the cheaper detection systems. Primers amplified a 123 base pairs fragment of the gene coding for glycoprotein D. PCR was tested on 25 samples of pig brain positive for AD and 85 samples negative in viral isolation and serum neutralization. The detection limit was calculated on samples of pig brain contaminated with a Brazilian isolate of SuHV-1. The technique had a detection limit of 10-0,5 TCID50/50µL. PCR was able to distinguish nonspecific reactions due to primer dimers (thus avoiding false positives) and get faster results.


Assuntos
Animais , Diagnóstico/métodos , Reação em Cadeia da Polimerase , Vírus/imunologia , Suínos/classificação
15.
Braz. j. microbiol ; Braz. j. microbiol;43(4): 1632-1640, Oct.-Dec. 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-665851

RESUMO

Suid herpesvirus 1 (SuHV-1) is the causative agent of pseudorabies (PR), a disease of great importance due to the huge losses it causes in the swine industry. The aim of this study was to determine a method for genotyping SuHV-1 based on partial sequences of the gene coding for glycoprotein C (gC) and to elucidate the possible reasons for the variability of this region. A total of 109 gCsequences collected from GenBank were divided into five major groups after reconstruction of a phylogenetic tree by Bayesian inference. The analysis showed that a portion of gC (approximately 671 bp) is under selective pressure at various points that coincide with regions of protein disorder. It was also possible to divide SuHV-1 into five genotypes that evolved under different selective pressures. These genotypes are not specific to countries or continents, perhaps due to multiple introduction events related to the importation of swine.


Assuntos
Animais , Variação Genética , Glicoproteínas/genética , Herpesvirus Suídeo 1/genética , Herpesvirus Suídeo 1/patogenicidade , Pseudorraiva/genética , Sequência de Bases/genética , Varicellovirus/genética , Varicellovirus/patogenicidade , Genética Microbiana , Genótipo , Métodos , Virulência
16.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);63(2): 296-301, abr. 2011. ilus, tab
Artigo em Português | LILACS | ID: lil-591118

RESUMO

Comparou-se a técnica nested PCR (nPCR) com os testes sorológicos IDGA e ELISA para o diagnóstico da anemia infecciosa equina. Amostras do DNA provenientes das células mononucleares do sangue periférico foram submetidas à amplificação do gene gag pela nPCR, que apresentou valores de sensibilidade e especificidade relativas de 90 por cento e 52,9 por cento, respectivamente, em relação à IDGA, e valores de 85,7 por cento e 49 por cento, respectivamente, em relação ao ELISA. Considerando-se os fatores referentes às limitações de cada técnica, pode ser sugerido o uso da nPCR como teste de diagnóstico complementar para AIE em amostras brasileiras.


The nested polymerase chain reaction (nPCR) technique was compared to AGID and ELISA serological tests for the diagnosis of Equine Infectious Anemia. DNA samples from the peripheral blood mononuclear cells were subjected to the amplification of the gag gene by nPCR, which showed relative sensibility and specificity values of 90.0 percent and 52.9 percent respectively, compared to the AGID and values of 85.7 percent and 49.0 percent, respectively, as compared to ELISA. Considering the factors concerning the limitations of each technique, the use of nPCR can be suggested as a complementary diagnostic test for EIA in Brazilian samples.


Assuntos
Animais , Anemia Infecciosa Equina/transmissão , Reação em Cadeia da Polimerase , Ensaio de Imunoadsorção Enzimática , Sorologia/tendências
17.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);63(6): 1405-1413, dez. 2011. ilus, tab
Artigo em Português | LILACS | ID: lil-608963

RESUMO

Desenvolveu-se uma PCR multiplex (mPCR) para diagnóstico diferencial de encefalite bovina causada por herpesvírus suíno 1 (SuHV-1), herpesvírus bovino 1 (BoHV-1), herpesvírus bovino 5 (BoHV-5) e herpesvírus ovino 2 (OvHV-2). Os iniciadores foram projetados após alinhamento de sequências disponíveis no banco de genomas (GenBank) e a reação foi padronizada levando-se em consideração a concentração dos reagentes e os tipos diferentes de DNA polimerase. Após determinação da especificidade e sensibilidade, 65 amostras de encéfalo de bovinos com síndrome neurológica foram submetidas à análise. A sensibilidade analítica para detecção de BoHV-1, BoHV-5 e SuHV-1 foi, respectivamente, 10(1,2) TCID50/50µL, 10(1,0) TCID50/50µL, 10(1,3) TCID50/50µL na reação multiplex. Das 65 amostras analisadas, 10 foram positivas para BoHV-5, uma para BoHV-1 e cinco para OvHV-2. A mPCR descrita neste trabalho mostrou-se uma técnica útil para o diagnóstico diferencial de enfermidades relacionadas ao sistema nervoso central de bovinos.


The aim of this study was to develop a multiplex PCR (mPCR) for the differential diagnosis of bovine encephalitis caused by the suid herpesvirus 1 (SuHV-1), bovine herpesvirus 1 (BoHV-1), bovine herpesvirus 5 (BoHV-5) and ovine herpesvirus 2 (OvHV -2). The primers were designed after alignment of sequences available in GenBank and the reaction was developed by taking into account the concentration of reagents and different types of DNA polymerase. After determining the specificity and sensitivity to PCR, 65 brain samples from cattle with neurological syndrome were submitted to the reaction. The analytical sensitivity for detection of BoHV-1, BoHV-5 and SuHV-1 was, respectively, 10(1,2) TCID50/50µL, 10(1,0) TCID50/50µL, 10(1,3) TCID50/50µL. Ten samples were positive for BoHV-5, one for BoHV-1, one for SuHV-1 and five for OvHV-2. The mPCR described here is a useful technique for the differential diagnosis of diseases related to the central nervous system of cattle.

18.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);62(5): 1259-1262, out. 2010. ilus
Artigo em Português | LILACS | ID: lil-570488

RESUMO

A duplex PCR was developed to differentiate the wild-type virus from the attenuated virus used in vaccinations. The PCR was able to amplify fragments of 493bp for glycoprotein E (gE) gene and 207bp for glycoprotein B (gB) gene. The analytical sensitivity was determined by addition of a virus field sample titled in the brain samples of pigs. The standard virus strain Shope, the vaccine strain Bartha, and ten other field isolates were subjected to PCR. The PCR was able to amplify fragments of gE and gB in all field samples and only fragments of gB were amplified in the attenuated virus, as expected. The technique was able to detect up to 100.5 TCID50/50mL virus in samples of brain. Duplex PCR proved to be an important tool for differentiation of naturally-infected animals and animals vaccinated with the virus deleted for gE.


Assuntos
Animais , Herpesvirus Suídeo 1/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Suínos/virologia , Vacinas
19.
Artigo em Inglês | MEDLINE | ID: mdl-12420867

RESUMO

Analysis of the partial bovine leukaemia virus (BLV) env gp51 gene sequences obtained from three BLV strains isolated in three different regions of Brazil was carried out. The Brazilian BLV env gp51 sequences were compared with seven other corresponding sequences of BLV strains isolated in different countries and with consensus sequence as well. The obtained data point on qualitative and quantitative differences among the analysed strains as far as the occurrence of single point mutations is concerned. Two Brazilian strains show significantly higher mutation rate than other analysed strains. Amino acid analysis did not show, however, any substantial changes of the primary protein structure coded by well conserved region of BLV env gp51 gene. Based on the obtained data, the putative dendogram image of possible phylogenetic relations among the studied BLV strains is presented as well.


Assuntos
DNA Viral/genética , Produtos do Gene env/genética , Vírus da Leucemia Bovina/genética , Sequência de Aminoácidos , Animais , Brasil/epidemiologia , Bovinos , Leucose Enzoótica Bovina/epidemiologia , Leucose Enzoótica Bovina/virologia , Vírus da Leucemia Bovina/classificação , Vírus da Leucemia Bovina/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária
20.
Arq. bras. med. vet. zootec ; Arq. bras. med. vet. zootec. (Online);59(4): 939-942, ago. 2007. ilus
Artigo em Português | LILACS | ID: lil-462190

RESUMO

Investigou-se a ocorrência da infecção pelo vírus da imunodeficiência felina (FIV) e pelo vírus da leucemia felina (FeLV) em gatos domésticos, provenientes de dois abrigos, no município de Belo Horizonte. Amostras de sangue de 145 animais foram coletadas e testadas para detecção do FIV pela reação em cadeia da polimerase (PCR). Destas amostras, 40 foram testadas para o antígeno p26 de FeLV por meio de ELISA. Observaram-se duas fêmeas (1,4 por cento) e quatro machos (2,8 por cento) positivos para FIV e nove fêmeas (22,5 por cento) e quatro machos (10,0 por cento) positivos para FeLV


The occurrence of the feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) was investigated in domestic cats from two shelters of Belo Horizonte. Samples from 145 cats were collected and tested for FIV by the polymerase chain reaction (PCR). Forty out of 145 samples were tested for FeLV p27 antigen by a commercial ELISA kit. Two females (1.4 percent) and four males (2.8 percent) were positive for FIV. For FeLV tests, 13 cats (32.5 percent) were positive, being nine females (22.5 percent) and four males (10.0 percent)


Assuntos
Animais , Gatos , Ensaio de Imunoadsorção Enzimática , Gatos/imunologia , Gatos/sangue , Reação em Cadeia da Polimerase/métodos , Vírus da Imunodeficiência Felina/isolamento & purificação , Vírus da Leucemia Felina/isolamento & purificação
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