Associação da técnica de cultivo em camada delgada contendo ágar Middlebrook 7H11 Modificado com a Reação em Cadeia de Polimerase (PCR) para identificação precoce de Mycobacterium bovis em órgãos de bovinos e bubalinos oriundos de abatedouros comerciais / Association between the modified Middlebrook 7H11 agar thin layer cultivation technique with the Polymerase Chain Reaction (PCR) on the earlier identification of Mycobacterium bovis in bovine and buffalo organs deriving from commercial slaughterhouses
AUTOR(ES)
Tatiana Reis do Rosário
DATA DE PUBLICAÇÃO
2010
RESUMO
The modified thin layer Middlebrook 7H11 cultivation was compared to the standard culture technique containing Stonebrink media, in order to evaluate the sensitivity and time for the detection of Mycobacterium bovis in bovine and buffalo organs deriving from commercial slaughterhouses. The PCR was applied for confirmation of the growth observed in the cultures, as well as the Ziehl-Neelsen color technique in order to detect acid-fast bacilli. 49 samples were submitted to the classic Petroffs decontamination method, and worked in two stages. In the first one, all of them were sowed in plates containing modified Middlebrook 7H11 agar in thin layer and in tubes containing the Stonebrink media, and the colonies observed macroscopically in both medias were submitted to the PCR. In the second stage, 10 samples from the first stage were submitted to a new culture, only in thin layer, for microscopical growth observation of the colonies and also submitted to the PCR. The results showed that: 1) Mycobacterium bovis cultivation in modified thin layer Middlebrook 7H11 for bovine and buffalo samples were viable when compared to the classic culture in Stonebrink, reducing the time of isolation, and can be applied as a complement to the traditional bovine tuberculosis diagnosis methods; 2) earlier identification of mycobacteria growth was found in thin layer (between 12nd and 25th day of growth) when compared to the Stonebrink; 3) PCR technique showed to be a complementary tool to the sum of techniques including Petroff decontamination, Ziehl-Neelsen color and thin layer cultivation for the diagnosis confirmation of the presence of mycobacteria in bovine and buffalo organs samples .
ASSUNTO(S)
mycobacterium bovis mycobacterium bovis buffalo organs camada delgada Órgãos bovinos e bubalinos polymerase chain reaction reação em cadeia de polimerase thin layer cultivation tuberculose tuberculosis
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