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. 1990 Nov;34(11):2184–2192. doi: 10.1128/aac.34.11.2184

Discrimination of extended-spectrum beta-lactamases by a novel nitrocefin competition assay.

G A Papanicolaou 1, A A Medeiros 1
PMCID: PMC172021  PMID: 2073109

Abstract

We describe a nitrocefin competition assay for determining inhibition profiles as a useful adjunct to existing biochemical methods for the discrimination of beta-lactamases. The hydrolysis rate of nitrocefin was measured with a plate photometer as the change in A480 over 45 min in the presence of 17 inhibitors. Fourteen well-established beta-lactamases and 13 extended-spectrum beta-lactamases were tested. Correlations with data from isoelectric focusing and amino acid sequencing suggested that the inhibition profile reflects alterations in the active-site configuration of beta-lactamases. The method was especially useful in measuring the relative affinities of beta-lactamases against poorly hydrolyzed substrates and in screening large numbers of isolates for the detection of new beta-lactamase types.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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