JAC Advance Access published online on August 4, 2004
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkh389
© 2004 by The British Society for Antimicrobial Chemotherapy
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1 Department of Microbiology, Jawaharlal Nehru Medical College and Hospital, Aligarh Muslim University, Aligarh--202002, Uttar Pradesh, India
* To whom correspondence should be addressed. E-mail: shahidsahar{at}yahoo.co.in.
Objectives: To develop an easy, rapid and reproducible spot-inoculation method for phenotypic detection of extended-spectrum Methods: ESBL and AmpC producing and non-producing isolates of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, as identified by the conventional three-dimensional extract test, were used to evaluate the modified procedures. Whole bacterial cells and freeze-thaw preparations, as Results: All the methods adopted for ESBL and AmpC detection by using the whole bacterial cells gave positive results. However, the best results were given by the spot-inoculation method. In modifications using the enzymic extracts, the enhanced growth of surface organisms was better appreciated in the designed modifications compared with the conventional methods. Conclusions: The method described here is simple and cost-effective. Furthermore, up to 16 isolates may be tested on a single culture plate, thus it is a less labour-intensive and more economic technique than other reported phenotypic methods.
Revised June 20, 2004
Accepted July 1, 2004
Brief report
Phenotypic detection of extended-spectrum and AmpC
-lactamases by a new spot-inoculation method and modified three-dimensional extract test: comparison with the conventional three-dimensional extract test
Abstract 
-lactamases (ESBLs) and AmpC -lactamases and to make the existing three-dimensional extract test more convenient for use in routine diagnostic laboratories.-lactamase sources, were strategically applied to culture plates near ceftazidime and cefoxitin discs on a lawn inoculum of E. coli ATCC 25922. Technical variations of the test included placing the -lactamase-containing inoculum into slits, wells and trenches, or onto the surface as spots at varying distances from the discs, and adding clavulanate or cloxacillin to the three-dimensional inoculum to confirm the presence of ESBLs and AmpC -lactamases, respectively.
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