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Journal of Antimicrobial Chemotherapy (2003) 52, 629-635
© 2003 The British Society for Antimicrobial Chemotherapy

AmpC cephalosporinase hyperproduction in Acinetobacter baumannii clinical strains

Stéphane Corvec1,*, Nathalie Caroff2, Eric Espaze1, Cécile Giraudeau1, Henri Drugeon1 and Alain Reynaud2

1 Laboratoire de Bactériologie-Virologie, Hygiène hospitalière, CHU, Nantes; 2 Laboratoire de Bactériologie-Virologie, Faculté de Pharmacie, Université de Nantes, Nantes, France

Received 16 April 2003; returned 21 May 2003; revised 7 July 2003; accepted 9 July 2003

Objective: To compare the genetic environments of ampC genes in different Acinetobacter baumannii isolates showing different levels of ß-lactam resistance.

Methods: The patterns of ß-lactam resistance and ß-lactamase production were investigated for 42 A. baumannii clinical strains. The MICs of various ß-lactams were determined in the presence or absence of the class C cephalosporinase inhibitor, cloxacillin (500 mg/L). The ampC gene and its 5' adjacent sequence were analysed by PCR and DNA sequencing. An RT–PCR method was developed to evaluate ampC transcript levels.

Results: Strains fell into three resistance groups: first, strains with a ceftazidime MIC <= 8 mg/L (20 strains, 47.6%); secondly, strains with a ceftazidime MIC 32 mg/L, which was reduced four-fold in the presence of cloxacillin (eight strains, 19%); and thirdly, strains with a ceftazidime MIC >= 256 mg/L, which did not decrease in the presence of cloxacillin (14 strains, 33.4%). In all of the resistant isolates (groups II and III), but not in any of the ceftazidime-susceptible isolates (group I), a 1180 bp insert showing all the characteristics of an insertion sequence was detected upstream from the ampC gene. Isolates having this insert overexpress ampC, according to RT–PCR experiments.

Conclusion: Presence of an insertion sequence upstream of ampC in A. baumannii clinical isolates, possibly including a strong promoter, has the potential to cause over-expression of AmpC, resulting in high-level ceftazidime resistance.

Keywords: AmpC hyperproducers, insertion sequences, Acinetobacter baumannii

* Correspondence address. Laboratoire de Bactériologie-Virologie-Hygiène hospitalière, CHU de Nantes – 9 quai Moncousu, 44093 Nantes Cedex 01, France. Tel: +33-2-40-08-39-55; Fax: +33-2-40-08-38-29; E-mail: stephane.corvec{at}chu-nantes.fr


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