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

• Main Authors: Zanini MS, Moreira EC, Lopes MTP, Oliveira RS, Leão SC, Fioravanti RL, Roxo E, Zumarraga M, Romano MI, Cataldi A, Salas CE
• Format: Article
• Language: English
• Published: Instituto Oswaldo Cruz, Ministério da Saúde 2001-01-01
• Series: Memórias do Instituto Oswaldo Cruz.
• Subjects: Mycobacterium bovis diagnosis; DNA polimorphism; thiocyanate guanidinium; Brazil
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762001000600012
• ISSN: 0074-0276; 1678-8060

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.