Shortening the incubation time for the combination disk diffusion extended-spectrum β-lactamase (ESBL) confirmation test: how far can we go?

- The combination disk diffusion ESBL confirmation test (CDT) is used to confirm ESBL production in Enterobacteriaceae.
- Concordance between 10-h and 18-h readings of the CDT is high.
- No change in the testing algorithm is needed.
- Turnaround time can be significantly reduced by early disk zone reading.

The combination disk diffusion extended-spectrum β-lactamase (ESBL) confirmation test (CDT) is used for the confirmation of ESBL production in Enterobacteriaceae and usually takes 16-20 h to results. In this study, we searched for the shortest possible incubation time without a reduction in reliability. A total of 125 ESBL screening-positive isolates were subjected to CDT and were molecularly characterised by microarray. Inhibition zones were read every hour over 6-18 h of incubation. Concordance between earlier and 18-h readings was calculated for each hour. Results were validated on 224 isolates during routine clinical practice. For the initial 125 isolates, concordance (Cohen's κ) between the 6-h and 18-h readings was 0.88 [95% confidence interval (CI) 0.78-0.96; P <0.001]. The earliest time point for full concordance with the 18-h reading was 10 h. Validation of the 10-h reading for 224 clinical isolates resulted in a concordance of 0.99 (95% CI 0.98-1.0) between the 10-h and 18-h readings. Overall concordance on all 349 isolates was 0.99 (95% CI 0.97-1.0). Reading after 10 h of incubation has an excellent correlation with results after 18 h of incubation. This can significantly reduce the turnaround time for ESBL detection in laboratories with long opening hours or providing a 24/7 service. Consequently, there is a potential for implementing infection control measures up to 8 h earlier.