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Early identification of mycobacterial species is crucial for early diagnosis. PCR-multiplex method performed on randomly chosen 54 mycobacteria isolates originating from clinical samples was found to be an inexpensive, quick and reliable alternative for commercially available diagnostics tests. Although the results of gene probes identification performed by NTLDR were generally consistent with multiplex PCR, two mixed Mycobacterium bovis BCG/Mycobacterium tuberculosis infections and a single misdiagnosis of M. tuberculosis with M. bovis were found. The routine application of multiplex-PCR has the potential to make diagnostics surveillance studies feasible.
The capacities of differentiation of Mycobacterium bovis BCG from other members of M. tuberculosis complex species using PCR-RFLP, multiplex PCR, and PCR-based genomic deletion analysis approaches were compared. In the study, mycobacteria isolated from patients suspected of adverse events following vaccination with BCG, primarily classified according presence of RD1 marker as virulent and avirulent mycobacteria, were used. The PCR-based genomic deletion analysis was found the best option for mycobacteria diagnostics improvement, as it was capable precisely differentiate virulent and avirulent mycobacteria or virulent species of M. tuberculosis complex. The routine confirmation of mycobacteria species in the cases of adverse events following BCG vaccination is highly expected, especially in clinical practice of patients with primary immunodeficiency.
In the study, phenotypic and genotypie properties of 42 Corynebacterium pseudotuberculosis strains isolated from 16 goat flocks were determined by random amplified polymorphic DNA and amplified fragment length polymorphism techniques. Several sets of RAPD primers and AFLP protocols were experimentally evaluated for potential of generating amplification profiles of the best discriminatory power. The overall similarity level of profiles by cluster analysis generated with RAPD3 (64%) set was higher than obtained for both RAPD 2 (70%) and AFLP (88%). The results confirmed usefulness of both AFLP and RAPD methods to discriminate strains of C. pseudotuberculosis, even at the flocks' level, and confirmed a single genotype as the most prevalent in goats coming from flocks in Poland.
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