Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex

Abstract

We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

Direct DNA Modified CTAB Preparation from Nasal Exudate in Live M. bovis Infected Cattle in Mexico Provide with a Valuable Assay Extrapolated to Humans TB Diagnostic Test

“…Detection, identification and differentiation of members of the MTBC complex rely in specificity, sensivity and accuracy of the methods that have been developed since the decades of the 90's. Despite this, still in endemic areas developing countries tuberculin field test as well as conventional techniques (histopathology and bacteriology) are performed due primarily to the costs and availability [25][26][27][28][29][30][31]. Therefore, it is a urgent need for endemic regions to have a routine assay to boost field test (false positive and negative tests) in live cows [37].…”

“…Therefore, it is a urgent need for endemic regions to have a routine assay to boost field test (false positive and negative tests) in live cows [37]. Moreover, the development of an optimal PCR methods that can boost field tuberculin tests (false positive/false negative) of live cows while avoiding unnecessary sacrifice of animals a molecular method should fulfill some basic requirements such as the high quality DNA, the regions to be amplified and primers design [8,16,22,[24][25][26][27][28][29][30][31][37][38][39]. M. bovis detection either from cattle or humans samples have moderate sensitivity, mainly attributed to the difficult for DNA extraction (bacterial lysis, long manipulation steps) or quality of the sample collection [17,[25][26][27][28].…”

“…DNA preparation from these colonies were made using filter column as indicated by the manufacturer. A thiny band of the amplicon (around 7 kb) was obtained from tissue ( Figure 3III-A, (lanes 1-4) or nasal exudate ( Figure 3III-A, lanes 5-7) [25,[35][36]. RD4 amplification of M. bovis DNA obtained from nasal exudate or from tissue was made and one fragment of around 400 pb was obtained in either case, tissue ( Figure 3III-B, lanes 1-4) or exudate ( Figure 3III-B, lanes 5-7).…”

“…The authors concluded that the PCR technique has a better analytical performance than the traditional culture for the detection of mycobacteria in nasal exudates of bovines [24]. To note is the several advancements for the improvement of detection, differentiation of the members of the MTB complex [25][26][27][28][29][30][31], each other have provided with a key point to take in the account either for TBb or human tuberculosis [25][26][27][28][29][30][31].…”

Identification of HPV Types and Mycobacterium Tuberculosis Complex in Historical Long-Term Preserved Formalin Fixed Tissues in Different Human Organs

“…bovis from other species of the MTB complex by a single point mutation [29]. At nucleotide position 169 of the pncA gene a change of C to G occurs in M .…”