Molecular Techniques for Identification of Species of the Mycobacterium tuberculosis Complex: The use of Multiplex PCR and an Adapted HPLC Method for Identification of Mycobacterium bovis and Diagnosis of Bovine Tuberculosis

Application of High Performance Liquid Chromatography for Identification of Mycobacterium spp

“…Also this method is relatively inexpensive and has been found to be more rapid alternative laboratory technique than the use of commercial nucleic acid probes [20]. Different methods have been developed for the detection of mycobacteria in clinical samples (e.g., blood, sputum) but they can also be applied to detection in other sources such as water [21] and milk [22]. Standard procedures to HPLC identification of mycobacterial species and most common steps used for different researchers are showed in the Figure 2 and Table 1, respectively.…”

“…However, they obtained chromatograms quite similar to those from the CDC protocol. On the other hand, Figueiredo et al [22] use a C-18 column 33% taller than those used in the CDC protocol (7.5 cm), increased the run time to 20 min. With these changes they observed a superior resolution in an adapted protocol and could be an alternative to discriminate between species with homologous HPLC chromatogram patterns.…”

Isolation and molecular characterization of Mycobacterium tuberculosis complex isolated from raw milk in some dairy farms in Egypt

“…In Nigeria, [40] indicated that the presence of M. tuberculosis in milk indicating post milking contamination from human and unhygienic practices associated with milking process. Furthermore, [41] detected M. bovis from (12%) of milk samples obtained from tuberculin positive cow cases. It is well established that cattle infected with M. bovis can excrete the bacillus in their milk.…”

Evaluation of the efficiency of nested q-PCR in the detection of Mycobacterium tuberculosis complex directly from tuberculosis-suspected lesions in post-mortem macroscopic inspections of bovine carcasses slaughtered in the state of Mato Grosso, Brazil

“…The DNA from the BTB-suspected tissue fragments was extracted using a Qiagen extraction and purification commercial kit (DNeasy® Blood & Tissue Kit), applying the protocol modifications as described by Figueiredo et al (2012). A 1 g tissue fragment was removed per animal.…”

“…Samples of lymph node tissue were spiked with the six concentrations of each plasmid DNA. DNA templates were prepared from the spiked samples using a Qiagen extraction kit (DNeasy® Blood & Tissue Kit), as described by Figueiredo et al (2012).…”