Identification of the Mycobacterium spp. Isolated from Cows Milk Samples by Using PCR Technique
The aim of this study was to identify the Mycobacterium isolates to species level by application of molecular method (polymerase chain reaction-PCR) with the specific primers that can differentiate the DNA of Mycobacterium tuberculosis and Mycobacterium bovis. Mycobacterium isolates used in this research were obtained from mastitic and apparently normal milk samples from many areas of Baghdad province in previous study. The isolates were previously identified as Mycobacterium based on direct identification of acid fast bacilli, cultural and biochemical properties. For differentiation of M.bovis from other species of Mycobacterium we used three sets of primers, these primers included a common forward primer 5'-TTCCGAATCCCTTGTGA-3', coded CSB1, reverse primer 5'- GGAGAGCGCCGTTGTA-3', coded CSB2 (M.bovis) and reverse primer 5'- AGTCGCGTGGCTTCTCTTTTA-3', coded CSB3 (M.tuberculosis). The m-PCR result showed that thirteen isolates out of fifteen was M.bovis as the product was 168bp and 2 isolates were M. tuberculosis complex as the product was 337bp. This research indicates that the m-PCR method using specific primers (CSB1, CSB2, and CSB3) can be applied for identification of M. tuberculosis and M. bovis and the importance of the implementation of measures for the prevention of tuberculosis in both humans and animals.
Mycobacterium tuberculosis, M. bovis, Milk, Multiplex Polymerase Chain Reaction (m-PCR), Cow
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