Cinomose em cães naturalmente infectados: técnicas diagnósticas e análise filogenética do gene da hemaglutinina do vírus da cinomose
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Data
2018-10-09
Autores
Santos, Romeu Moreira dos [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A cinomose canina é uma das enfermidades infectocontagiosas mais importantes que acomete os cães. É uma doença causada pelo vírus da Cinomose (“Canine Distemper Virus” - CDV), um Paramyxovirus, do gênero Morbillivirus, de ocorrência mundial, sem sazonalidade, sem predileção de sexo ou raça, apresenta maior incidência em animais jovens, podendo acometer todas as idades. No Brasil, pesquisas sobre o uso de técnicas que comparam os diferentes testes diagnósticos direto para o vírus da cinomose (CDV) associado a análise molecular e filogenética do mesmo ainda continuam escassas. Além disso, são poucos os estudos sobre epidemiologia molecular que relatam diferenças marcantes na análise filogenética do gene hemaglutinina (H) do CDV entre os isolados de campo e as cepas de referência do CDV, especialmente àquelas utilizadas na produção de vacinas. Dessa maneira, o presente trabalho tem como principal objetivo avaliar os principais métodos diagnósticos laboratoriais para identificação do CDV em cães com suspeita de cinomose associando à análise filogenética do gene H dos isolados de campo. O Ensaio imunocromatografico direto (IC), a RT-PCR e a Nested-PCR foram avaliados para detecção do CDV em diferentes amostras clínicas (urina, suabes retais e conjuntivais) de 62 animais suspeitos de cinomose, provenientes do atendimento ambulatorial do hospital veterinário da Unesp (Jaboticabal-SP) e de clínicas particulares da região. Após detecção viral as amostras foram submetidas a sequenciamento seguida de análise filogenética do gene H do CDV. Um total de 42 animais foram positivos em pelo menos uma das técnicas utilizadas neste experimento. As técnicas moleculares (RT-PCR e Nested-PCR) demonstraram maior sensibilidade que o ensaio comercial de IC. Os resultados obtidos da análise filogenética do gene H sugeriram que o grupo de isolados do CDV no Brasil são distintos das cepas vacinais e são mais relacionados genotipicamente aos isolados Europeus 1/Sul-Americanos 1. Em conclusão, a técnica Nested-PCR aplicada na detecção do CDV em suabes retais revelou-se o método mais eficaz e sensível para identificação desse vírus, que associado a análise filogenética pode ser classificado em um clado diferente das cepas vacinais sugerindo um potencial de infecção para os cães domésticos inclusive os vacinados.
Canine distemper is one of the most important infectious diseases that affects dogs. It is a Canine Distemper Virus (CDV), a Paramyxovirus, of the genus Morbillivirus, a global deforest, without seasonality, without predilection of sex or race, that is constituted by the majority of the animals, being able to affect all ages. In Brazil, research on the use of techniques that compare the different direct diagnostic tests for the distemper virus (CDV) associated with a molecular and phylogenetic analysis of the same are still scarce. In addition, there are some studies on molecular epidemiology that report differences in the phylogenetic analysis of the hemagglutinin (H) gene from CDV between the fields of action and as a reference for CDV, especially the practices used in the production of vaccines. Thus, the main objective of this study is to identify laboratory diagnoses for the identification of CDV in dogs with suspected pair association to the phylogenetic analysis of the H gene of the field isolates. The direct immunochromatographic (CI) assay, RT-PCR and nested-PCR were evaluated for the detection of CDV in different animal species (urine, swabs and conjunctival) of 62 suspected cases of distemper from the outpatient clinic of the Unesp (Jaboticabal-SP) and the private medical records of the region. After viral detection as sequential sequencing followed by phylogenetic analysis of the CDV H gene. A total of 42 animals were positive at least one of the experiment techniques in this experiment. As molecular techniques (RT-PCR and Nested-PCR) demonstrated greater sensitivity than the commercial IC test. The results obtained from the phylogenetic analysis of the H gene suggest that the group of CDV cells in Brazil are distinct from the vaccine strains and are more commonly genotyped to the following European 1 / South American 1. In conclusion, a Nested-PCR technique. CDV can be used as a more effective and sensitive method to virus infection, which is associated with a phylogenetic analysis that can be classified into one of the different clones of the vaccines, suggesting a potential for infection for animals that include the vaccinated.
Canine distemper is one of the most important infectious diseases that affects dogs. It is a Canine Distemper Virus (CDV), a Paramyxovirus, of the genus Morbillivirus, a global deforest, without seasonality, without predilection of sex or race, that is constituted by the majority of the animals, being able to affect all ages. In Brazil, research on the use of techniques that compare the different direct diagnostic tests for the distemper virus (CDV) associated with a molecular and phylogenetic analysis of the same are still scarce. In addition, there are some studies on molecular epidemiology that report differences in the phylogenetic analysis of the hemagglutinin (H) gene from CDV between the fields of action and as a reference for CDV, especially the practices used in the production of vaccines. Thus, the main objective of this study is to identify laboratory diagnoses for the identification of CDV in dogs with suspected pair association to the phylogenetic analysis of the H gene of the field isolates. The direct immunochromatographic (CI) assay, RT-PCR and nested-PCR were evaluated for the detection of CDV in different animal species (urine, swabs and conjunctival) of 62 suspected cases of distemper from the outpatient clinic of the Unesp (Jaboticabal-SP) and the private medical records of the region. After viral detection as sequential sequencing followed by phylogenetic analysis of the CDV H gene. A total of 42 animals were positive at least one of the experiment techniques in this experiment. As molecular techniques (RT-PCR and Nested-PCR) demonstrated greater sensitivity than the commercial IC test. The results obtained from the phylogenetic analysis of the H gene suggest that the group of CDV cells in Brazil are distinct from the vaccine strains and are more commonly genotyped to the following European 1 / South American 1. In conclusion, a Nested-PCR technique. CDV can be used as a more effective and sensitive method to virus infection, which is associated with a phylogenetic analysis that can be classified into one of the different clones of the vaccines, suggesting a potential for infection for animals that include the vaccinated.
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Palavras-chave
Canis lupus familiaris, Detecção viral, Filogenia, Morbillivirus