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Journal of Antimicrobial Chemotherapy (1993) 31, 841-854
© 1993 The British Society for Antimicrobial Chemotherapy

other

The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other Gram-negative bacteria

A. M. Snellinga, P. M. Hawkeya, J. Heritagea, P. Downeyb and B. Homesc

aDepartment of Microbiology, University of Leeds Leeds LS2 9JT, UK bDepartment of Pathology and Microbiology, University of Bristol, University Walk Bristol BS8 1TD, UK cNational Collection of Type Cultures, Central Public Health Laboratory, 61 Colindale Avenue London NW9 5HT, UK

Received 5 October 1992; accepted 18 January 1993

A nucleotide probe for the chromosomal aminoglycoside 6' acetyltransferase gene (aac(6')-Ic) of Serratia marcescens was used in non-radioactive dot-blot hybridization experiments with 186 strains belonging to ten different species of Serratia. The gene was only detected in strains of S. marcescens (all strains tested), and positive hybridization was seen irrespective of whether or not strains were kanamycinresistant. An additional 180 strains belonging to 28 Gram-negative bacterial species other than in the genus Serratia did not hybridize with the probe. A rapid PCR test for the aac(6')-Ic gene was developed and used to confirm that the aac(6')-Ic gene is only found in S. marcescens. Southern hybridization analysis of S. marcescens chromosomal DNA demonstrated that the gene was usually located on a PvuII fragment with a putative isoleucine tRNA-2 gene, but polymorphisms with respect to the size of this fragment were observed.


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