Isolamento do vírus ectima contagioso e identificação pela reação em cadeia da polimerase. / Contagious ecthyma virus isolation and identification by polymerase chain reaction.

AUTOR(ES)

Jailson Honorato Pinto Júnior

FONTE

IBICT - Instituto Brasileiro de Informação em Ciência e Tecnologia

DATA DE PUBLICAÇÃO

31/08/2011

RESUMO

Contagious ecthyma (CE) is an acute viral disease that affects sheep and goats, widespread throughout the world, including Brazil, especially in the Northeast, where sheep and goat rearing is practiced for the production of skin, meat and milk. In the state of Pernambuco, has been reported as one of the major infectious diseases of goats and sheep in semi-arid. Despite the endemicity of the disease and its importance, few research studies have been conducted in the country with the virus CE, that can support the control of the disease, grounded mainly on vaccination of animals. CE is a disease that can be confused with the vesicular such as Footh and Mouth Disease, with the need for differentiation, especially to support the actions of the National Program for the Eradication of Footh and Mouth Disease (PNEFA) of the Ministry of Agriculture and Livestock (MAPA). For this it is necessary to have a fast and efficient test for diagnosis. Thus aimed to isolate the virus from clinical specimens CE, as well as confirm the etiology of the disease by Polymerase Chain Reaction (PCR). Crust samples collected from seven animals originating in the state of Pernambuco, with clinical signs suggestive of CE, were inoculated in corneal epithelial cells fetal goats (FCC 40) and evaluated every 24 hours for observation of cytopathic effect (CPE). The supernatant layers were frozen and processed for the extraction of viral DNA to be used in PCR, using the primers for B2L gen amplification. The sample studied CE virus had varying degrees (10 to 40%) of CPE in cultures of corneal cells, with most featuring CPE in only about 10%. The CPE was characterized by cells that have become uneven in appearance, rounded appearance of syncytia and vacuoles, shrinkage of cell membranes of neighboring cells and detachment of cells. The specificity of CPE was confirmed by PCR, with amplification of DNA fragments with 235 bp, which corresponds to the expected size. It is concluded that the EC virus isolation in cell cultures FCC 40 and its identification by PCR is a method applicable to the diagnosis of CE potentially useful in the differential diagnosis of vesicular diseases of PNEFA.

ASSUNTO(S)

orf isolamento diagnóstico pcr medicina veterinaria isolation diagnosis

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