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Validação e aplicação da RT-QPCR de RNAm codificantes de citocinas em gatos naturalmente infectados pleo coronavírus felino (FCOV)

Authors: Zadra, Vívian Ferreira;

Validação e aplicação da RT-QPCR de RNAm codificantes de citocinas em gatos naturalmente infectados pleo coronavírus felino (FCOV)

Abstract

O vírus da peritonite infecciosa felina (FIPV) causa uma doença fatal, sistêmica e imunomediada. O FIPV contem mutações do coronavírus entérico felino (FECoV) e o diagnóstico antemortem da PIF é desafiador pois a maioria dos gatos são positivos para FCoV nos testes sorológicos e de amplificação de DNA. A diferenciação entre a infecção pelo FIPV e pelo FECoV é difícil, sendo conclusivo post-mortem pelo exame histopatológico. A RT-PCR para detecção do RNAm de coronavírus felino em sangue periférico é um teste diagnóstico que evidencia a infecção e replicação do vírus. Entretanto, esses ensaios não conseguem distinguir entre cepas produtoras de PIF e do FECoV. Na patogenia da infecção, as células imunes infectadas induzem uma produção exacerbada de citocinas. Assim, nosso objetivo foi o de verificar se gatos com PIF expressam RNAM codificante de citocinas de forma alterada quando comparadas com animais saudáveis. Para tanto, amostras de sangue de 44 gatos provenientes de gatis de SP e RJ foram coletadas em 4 momentos, num intervalo de 4 meses. A partir do RNA total extraído, foi realizada a RT-qPCR para RNAm codificante de citocinas felinas (IL-1, IL-2, IL-6, IL-10, IL-12, TNF-α, IFN-γ) normalizadas pelo GAPDH. Observou-se que, para as citocinas IL-1, IL-2, IL-6 e IL-10, foram obtidos resultados elevados na maioria dos animais. Entretanto, IL-12, TNF-α e IFN-γ estavam aumentadas em apenas alguns grupos, que pode estar associado à infecção por FIPV. Os animais que apresentaram valores elevados para essas citocinas, tinham sinais clínicos compatíveis com a doença e contato com animais que desenvolveram a PIF

The feline infectious peritonitis virus (FIPV) causes a fatal disease, and systemic immune mediated. The FIPV contains mutations of feline enteric coronavirus (FECoV) and antemortem diagnosis of FIP is challenging because most cats are positive for FCoV in serological and DNA amplification. The differentiation between infection the FIPV and FECoV is hard being conclusive post-mortem by histopathological examination. RT-PCR for detection of feline coronavirus mRNA in peripheral blood is a diagnostic test which shows the infection and replication of the virus. However, these tests can’t distinguish between strains producing FIP and FECoV. In the pathogenesis of infection, the infected cells induce an immune exacerbated production of cytokines. Thus, our goal was to determine whether cats with FIP express mRNA encoding cytokines altered form compared to healthy animals. Therefore, blood samples from 44 cats from catteries of SP and RJ were collected at 4 times within 4 months. From the total RNA, we performed RT-qPCR mRNA coding for feline cytokine (IL-1, IL-2, IL-6, IL-10, IL-12, TNF-α, IFN-γ) normalized by GAPDH. It was observed that, for the cytokines IL-1, IL-2, IL-6 and IL-10 high scores were obtained in most animals. However, IL-12, TNF-α and IFN-γ were increased only in some groups, which may be associated with FIPV infection. The animals with high values for these cytokines had clinical signs consistent with the disease and contact with animals that developed FIP

Pós-graduação em Medicina Veterinária - FMVZ

Country
Brazil
Keywords

Coronavírus, Peritonio - Inflamação, Peritonite, Cytokines, Citocinas, Felideo - Doenças

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
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