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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060 EISSN: 1678-8060
Vol. 96, No. 6, 2001, pp. 809-813
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Bioline Code: oc01130
Full paper language: English
Document type: Research Article
Document available free of charge
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Memórias do Instituto Oswaldo Cruz, Vol. 96, No. 6, 2001, pp. 809-813
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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
MS Zanini; EC Moreira; MTP Lopes; RS Oliveira; SC Leão; RL Fioravanti; E Roxo; M Zumarraga; MI Romano; A Cataldi & CE Salas
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 a -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.
Keywords
Mycobacterium bovis diagnosis, DNA polimorphism, thiocyanate guanidinium, Brazil
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