Direct Detection and Identification of Mycobacterium tuberculosis and Mycobacterium bovis in Bovine Samples by a Novel Nested PCR Assay: Correlation with Conventional Techniques

摘要: Mycobacterium tuberculosis and M. bovis infect animals humans. Their epidemiologies in developed developing countries differ, owing to differences the implementation of preventive measures (World Health Organization, 1999). Identification differentiation these closely related mycobacterial species would help determine source, reservoirs infection, disease burden due diverse pathogens. The utility hupB gene (Rv2986c tuberculosis, or Mb3010c bovis) differentiate was evaluated by a PCR-restriction fragment length polymorphism (RFLP) assay with 56 characterized bovine isolates (S. Prabhakar et al., J. Clin. Microbiol. 42:2724-2732, 2004). degree concordance between PCR-RFLP microbiological characterization 99.0% (P < 0.001). A nested PCR (N-PCR) developed, replacing for direct detection samples. N-PCR products corresponded 116 89 bp, respectively. limit DNA 50 fg, equivalent five tubercle bacilli. and/or detected 55.5% (105/189) samples N-PCR, compared 9.4% (18/189) culture. sensitivities culture were 97.3 29.7, respectively, their specificities 22.2 77.7%, percentages identified as infected reflected clinical categorizations cattle <0.05 <0.01). Mixed infection 22 animals, whereas mixed 1 animal.