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Byline: E.E. Stobberingh (1), J. Arends (2), J.A.A. Hoggkamp-Korstanje (3), W.H.F. Goessens (4), M.R. Visser (5), A.G.M. Buiting (6), Y.J. Debets-Ossenkopp (7), R.J. van Ketel (8), M.L. van Ogtrop (9), L.J.M. Sabbe (10), G.P. Voorn (9), H.L.J. Winter (11), J.H. van Zeijl (12) Keywords: Key words ESBL; Klebsiella; E. coli; Epidemiology Abstract: The prevalence of ESBL was determined among isolates of Escherichia coli (n = 571) and Klebsiella spp. (n = 196) collected during a 1-week study period in 8 university and 3 large regional laboratories all over the Netherlands. 18 isolates were positive for at least one of the screening tests used, i.e., VITEK-ESBL, E-test ESBL and MIC ratio of ceftazidime/ceftazidime-clavulanic acid, cefotaxime/cefotaxime-clavulanic acid. In 5 of these 18 putative ESBLs no betalactamase production was detectable. A TEM type was found in three E. coli and two Klebsiella spp. An SHV type was present in five Klebsiella spp. In one E. coli and one Klebsiella pneumoniae both enzymes were present. In one Klebsiella oxytoca neither of the two enzymes was present. Using PCR for both ESBL TEM and ESBL SHV, an SHV ESBL was found in one E. coli and four Klebsiella isolates. The mutations at position 238 and 240 were already described. In one E. coli isolate a TEM ESBL was found with three mutations, at position 21, 164 and 265. These mutations were already described in other ESBLs but not in this combination suggesting a new TEM ESBL. The overall prevalence of ESBL producing E. coli and Klebsiella spp. was less than 1% (6 out of 767). Author Affiliation: (1) University Hospital Maastricht, Medical Microbiology, P.O. Box 5800, NL-6202 AZ Maastricht, The Netherlands, Phone: +31-43-3874644, Fax: +31-43-3876643, e-mail: EST@LMIB.AZM.NL, NL (2) University Hospital Groningen, Medical Microbiology, P.O. Box 30001, 9799 RB Groningen, The Netherlands, NL (3) University Hospital Nijmegen, Medical Microbiology, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands, NL (4) Hospital Dijkzigt, Medical Microbiology, Dr. Molenwaterplein 40, 3033 CC Rotterdam, The Netherlands, NL (5) University Hospital Utrecht, Clinical Microbiology, P.O. Box 85500, 3508 GA Utrecht, The Netherlands, NL (6) Public Health Laboratory, P.O. Box 747, 5000 AS Tilburg, The Netherlands, NL (7) University Hospital Amsterdam, Clinical Microbiology, P.O. Box 7057, 1007 MB Amsterdam, The Netherlands, NL (8) Medical Centre Amsterdam, Medical Microbiology, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands, NL (9) University Hospital Leiden, Medical Microbiology, P.O. Box 9600, 2300 RC Leiden, The Netherlands, NL (10) Oosterschelde Hospital, Medical Microbiology, 's Gravenpolderseweg 114, 4462 RA Goes, The Netherlands, NL (11) Public Health Laboratory, van Ketwich Verschuurlaan 92, 9721 SW Groningen, The Netherlands, NL (12) Public Health Laboratory, P.O. Box 21020, 8900 JA Leeuwarden, The Netherlands, NL Article note: Received: December 14, 1998 * Accepted: September 19, 1999