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Journal of Antimicrobial Chemotherapy (1987) 20, 645-656
© 1987 The British Society for Antimicrobial Chemotherapy

research-article

Ampicillin resistance in Haemophilus influenzae: identification of resistance mechanisms

Alison J. Reid*, Iain N. Simpsona,{dagger}, Paul B. Harpera,{ddagger} and Sebastian G.B. Amyes§

Bacteriology Department, The Medical School, University of Edinburgh Teviot Place, Edinburgh EH8 9AG aChemotherapy Department, Microbiology Division, Glaxo Group Research Ltd Greenford Road, Greenford, UB6 OHE, U.K.

accepted 28 May 1987

§Corresponding author

The incidence and mechanisms of ampicillin resistance (MIC > 1 mg/l) were investigated in 105 clinical isolates of Haemophilus influenzae collected in Edinburgh during 1983/4. Fifteen (14.3%) ampicillin-resistant strains were identified and these were non-serotypable and comprised six biotypes. Isoelectric focusing and ß-lactamase-inhibition studies demonstrated that production of the TEM-1 ß-lactamase was the principal mechanism of resistance in nine (60%) strains. Radiolabelling revealed that one ß-lactamase-positive strain also had an unusual penicillin-binding protein (PBP) profit. No ß-lactamase activity was detected in the other six (40%) ampicillin-resistant strains. Two ß-lactamase-negative ampicillin-resistant strains had atypical PBP profiles. SDS-PAGE analysis showed that four ß-lactamase-negative ampicillin-resistant strains, including one with altered PBPs, exhibited outer membrane protein profiles which differed from those of sensitive strains of the same biotype. The ampicillin-resistance mechanism of the remaining strain could not be determined. Thus, several resistance mechanisms, either acting individually or in combination, are implicated in ampicillin resistance in H. influenzae.


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