Identification of Nontuberculous Mycobacteria in Patients with Pulmonary Diseases in Gyeongnam, Korea, Using Multiplex PCR and Multigene Sequence-Based Analysis

Figure 1

Validation of the two-step multiplex PCR using reference strains. (a) The first mPCR reaction was conducted with mycobacterial reference strains. The amplified products were of the following sizes: Lane M, DNA ladder; Lane 1, 358 bps (RD9), 484 bps (16s rRNA), and 705 bps (rv0577) for MTB NCCP 72077; Lane 2, 484 bps (16s rRNA) and 705 bps (rv0577) for M. bovis BCG Pasteur 1173P2; Lane 3, 282 bps (IS1311) and 484 bps (16s rRNA) for M. avium subsp. hominissuis 104; Lane 4, 282 bps (IS1311), 484 bps (16s rRNA), and 753 bps (IS901) for M. avium subsp. avium KBN12P06234; Lane 5, 106 bps (DT1) and 484 bps (16s rRNA) for M. intracellulare ATCC 13950; Lane 6, 484 bps (16s rRNA) for M. kansasii KBN12P06233; Lane 7, M. abscessus KCTC 19621; Lane 8, M. massiliense KCTC 19086; Lane 9, M. fortuitum KBN12P06244; and Lane 10, negative control. (b) The second mPCR reaction was conducted with mycobacterial reference strains. The amplified products were of the following sizes: Lane M, DNA ladder; Lane 1, 310 bps (mass_3210) for M. abscessus; Lane 2, 1145 bps (mass_3210) for M. massiliense; Lane 3, 275 bps (SOD) for M. fortuitum; Lane 4, 582 bps (transferase) for M. kansasii; and Lane 5, negative control.

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