Proposal of a new method for subtyping of Mycobacterium kansasii based upon PCR restriction enzyme analysis of the tuf gene

•In the presented study, on the basis of sequence analysis of the tuf gene, from representatives of the M. kansasii subtypes I–VI, we designed a new molecular tool for the subtype discrimination.•The proposed PCR restriction enzyme analysis assay (PCR-REA) should provide a clinically useful method of identifying M. kansasii subtypes since its application is relatively fast, inexpensive, and simple to use.•The main advantage of the proposed method over previously designed PCR-REA assays is the use of only 1 digestion enzyme (instead of 2 or 3, as in previously designed and currently used assays).

Within this study, a new, rapid method for subtyping of Mycobacterium kansasii was developed based on the sequence analysis of the tuf gene coding for the Tu (thermo-unstable) elongation factor (EF-Tu). The method involves PCR amplification of ca. 740-bp tuf gene fragment, followed by digestion with the MvaI restriction endonuclease.