Analysis Of The Mechanism Of Acinetobacter Baumanii Resistance To Carbapenems

Objective: Analyze carbapenemase and out membrane proteins of 5 Acinetobacter Baumanii isolates resistance to carbapenems to investigate the mechanism of Acinetobacter Baumanii resistance to carbapenems in Jinan.Methods: Extracted chromosome DNA of 5 Acinetobacter Baumanii isolates resistance to carbapenems determined by E-test was subjected to polymerase chain reaction (PCR) with the specific primers of bla_oxA-23(FR 5'-GATGTGTCATAGTATTCGTCG-3' , RV 5'-TCACAACAACTAAAAGCACTG-3' )> bla_oxA-24(FR 5' -TTCCCCTAACATGAATTTGT-3' , RV 5' -GTACTAATCAAAGTTGTGAA -3'), bla_IMP(FR 5' -CTACCGCAGCAGAGTCTTTAC -3' , RV 5' -AACCAGTTTTGCCTTACCAT -3' ) bla_VIM(FR 5'-TCCGACAGTCAGCGAAAT -3', RV 5' -GCACAACCACCATAGAGCA-3' ) . The conditions for the presence of bla_oxA-23-specific primers comprised 1 cycle at 94℃ for 5 min, followed by 30 cycles at 94℃ for 1 min, 52℃ for 1 min, and 72℃ for 2 min and a final elongation at 72℃ for 7 min. The conditions for the presence of bla_oxA-24-specific primers comprised 1 cycle of denaturation at 94℃ for 4 min, 30 cycles at 94℃ for 1 min, 48° C for 1 min, and 72℃ for 2 min and then a final elongation at 72° C for 10 min. The conditions for the presence of bla_IMP-specific primers and bla_VIM-specific primers comprised 1 cycle of denaturation at 94℃ for 5 min, 30 cycles at 94℃ for 30 sec, 56℃ for 30 sec, and 72° C for 45 sec and then a final elongation at 72℃ for 7 min. The PCR products were ligated into PMD18-T, transformed into DH5 α and then sequenced.Isolates were inoculated in nutrient broth, grown overnight at 36℃.