Genomic DNA was isolated using the Nucleospin Tissue DNA extraction kit (Machery Nagel,
Düren, Germany). Acquired AMR genes, which encode mechanisms of resistance to carbapenems
(blaOXA-23-like), tetracyclines [tet(B) and
tet(39)] and aminoglycosides (strA, strB, aac(3)-Iaand aac(6´)-Im) in Acinetobacter spp., were detected by
simplex PCRs (Supplementary Table 1 ). Additional aminoglycoside-modifying enzyme
(AME)-encoding genes including aac(3)-IIa, aac(6´)-Ih,
aph(3′)-VI, ant(2´´)-Ia, aph(3′)-Ia and
aac(6´)-Ib were identified by multiplex PCRs [30 (link)]. A 25-μL PCR mixture was prepared using the 5X Firepol Master Mix
(Solis BioDye, Tartu, Estonia) with 0.2 μM of each primer. Entire
blaOXA-23 were analyzed by Sanger’s capillary sequencing.
Specific regions of ISAba1 and ISAba4 at both
extremities of the blaOXA-23-containing transposon were
amplified by PCR to illustrate the presence of transposable elements [25 (link)].
Düren, Germany). Acquired AMR genes, which encode mechanisms of resistance to carbapenems
(blaOXA-23-like), tetracyclines [tet(B) and
tet(39)] and aminoglycosides (strA, strB, aac(3)-Iaand aac(6´)-Im) in Acinetobacter spp., were detected by
simplex PCRs (
(AME)-encoding genes including aac(3)-IIa, aac(6´)-Ih,
aph(3′)-VI, ant(2´´)-Ia, aph(3′)-Ia and
aac(6´)-Ib were identified by multiplex PCRs [30 (link)]. A 25-μL PCR mixture was prepared using the 5X Firepol Master Mix
(Solis BioDye, Tartu, Estonia) with 0.2 μM of each primer. Entire
blaOXA-23 were analyzed by Sanger’s capillary sequencing.
Specific regions of ISAba1 and ISAba4 at both
extremities of the blaOXA-23-containing transposon were
amplified by PCR to illustrate the presence of transposable elements [25 (link)].
