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Mycobacterium bovis: polymerase chain reaction identification in bovine Lymphonode biopsies and genotyping in isolates from Southeast Brazil by spolygotyping and restriction fragment length polymorphism

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Author(s): Zanini MS | Moreira EC | Lopes MTP | Oliveira RS | Leão SC | Fioravanti RL | Roxo E | Zumarraga M | Romano MI | Cataldi A | Salas CE

Journal: Memórias do Instituto Oswaldo Cruz
ISSN 0074-0276

Volume: 96;
Issue: 6;
Start page: 809;
Date: 2001;
Original page

Keywords: Mycobacterium bovis diagnosis | DNA polimorphism | thiocyanate guanidinium | Brazil

ABSTRACT
Diagnosis of the Mycobacterium tuberculosis complex by direct PCR of mediastinal lymphnode DNA and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. The PCR procedure applied on DNA samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70% of the samples. PCR confirmed the identification of 23 samples (100%) that grew in culture, 9 samples (60%) that failed to grow in culture, plus 6 (37.5%) samples that resulted in growth of bacterial contaminants. Genotyping by IS6110-RFLP and DR-spoligotyping analysis of seven samples revealed the presence of several polimorphisms. Seven of the isolates contained multiple copies of IS6110, thus defining the existence of five singular genotypes.
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