Detecção de DNA de Brucella abortus pela PCR em leite bubalino experimentalmente contaminado pela amostra 1119-3 / Detection of Brucella abortus DNA by PCR in spiked buffalo milk with B. abortus strain 1119-3

AUTOR(ES)

Maria Carolina Guido

DATA DE PUBLICAÇÃO

2003

RESUMO

To improve the PCR performance for detection of Brucella spp. in buffalo milk, different DNA extraction protocols were carried out in buffalo milk spiked with Brucella abortus strain 1119-3. These protocols were based on utilization of guanidine isothiocyanate, boil and proteinase K, with some variations. The analytical sensitivity was the evaluation criteria. Boiling with initial filtration of the sample through a solution of sacarosis 40% presented analytical sensitivity of 103 CFU/ml. The protocols based on proteinase K presented analytical sensitivity of 104 CFU/ml. The highest analytical sensitivity (10 CFU/ml) and best visualization of the amplified product were verified for the protocol using guanidine isothyocianate with immediate precipitation in alcohol without addition of Tween 20 in the initial sample wash.

ASSUNTO(S)

búfalos leite milk pcr pcr animal brucellosis buffalo brucelose animal

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