Mycobacteria Identification via Multiplex PCR

DNA isolation was performed using Genomic Mini AX Bacteria (A&A Biotechnology, Gdańsk, Poland) following the manufacturer’s instructions (https://www.aabiot.com/pobierz?code=15de0a1fe3fd35e7688cf0fe68a91af304ece5d2; accessed on 4 March 2023). To determine whether a given strain belonged to tuberculous or nontuberculous mycobacteria, multiplex PCR was performed (Table 1, Table 2 and Table 3), as previously described [16 (link)]. In this method, two pairs of primers were used, which allowed for the simultaneous amplification of two different DNA fragments [18 ]. The primers were constructed by the European Union Reference Laboratory for bovine tuberculosis in Madrid and synthesized by Eurofins Genomics company (Eurofins Genomics, Ebersberg, Germany).