Samples of the Enterobacter cloacae complex were
identified by amplification and sequencing of a fragment of
rpoB gene. PCR was performed in the same conditions
as described above, using the primers Vic3a (forward):
50-GGCGAAATGGCWGAGAACCA-30 and Vic2a (reverse):
50-GAGTCTTCGAAGTTGTAACC-30 [46]. The thermal cycle
consisted an initial denaturation step at 94 C for 1.5
minutes, followed by 40 cycles of 94 C/10 seconds, 55 C/
20 seconds and 72 C/50 seconds, with final extension of
72 C/5 minutes [47]. Amplified products were purified
using GFX PCR DNA and Gel Band Purification kit (GE
Healthcare) for subsequent sequencing using BigDye
Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems
Life Technologies) for an automated ABI sequencer 3500xL