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Journal of Clinical Microbiology, April 2005, p. 1604-1611, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1604-1611.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Discrimination within Phenotypically Closely Related Definitive Types of Salmonella enterica Serovar Typhimurium by the Multiple Amplification of Phage Locus Typing Technique

Ian L. Ross and Michael W. Heuzenroeder*

Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, South Australia, Australia

Received 21 September 2004/ Returned for modification 18 November 2004/ Accepted 6 December 2004

Multilocus sequence typing (MLST) is a relatively new high-resolution typing system employed for epidemiological studies of bacteria, including Salmonella. Discrimination based on MLST of housekeeping genes may be problematical, due to the high identity of gene sequences of closely related Salmonella species. The presence of genomic sequences derived from stable temperate phages in Salmonella offers an alternative for MLST of Salmonella. We have used MLST of prophage loci in Salmonella enterica serovar Typhimurium to discriminate closely related isolates of serovar Typhimurium. We have compared these results to MLST of five housekeeping genes, as well as pulsed-field gel electrophoresis (PFGE). The presence or absence of prophage loci in the 73 serovar Typhimurium isolates tested, as well as allelic variation as detected by sequencing, provided greater discrimination between isolates than either MLST of housekeeping genes or PFGE. Amplification of prophage loci alone separated serovar Typhimurium isolates into 27 groups comprising multiple isolates or individual strains. Sequencing of isolates found within the clusters separated isolates even further. By contrast, PFGE could only divide the 73 isolates into five distinct groups. MLST using housekeeping genes did not provide any significant separation of isolates in comparison to amplification or MLST of prophage loci. The results demonstrate that the amplification and sequencing of prophage loci provides a high-resolution, objective method for the discrimination of closely related isolates of serovar Typhimurium. It is proposed that multiple amplification of phage locus typing may provide sufficient discrimination for epidemiological purposes without recourse to MLST.

* Corresponding author. Mailing address: Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Box 14, Rundle Mall, P.O. Adelaide, S.A. 5000, Australia. Phone: 618 8222 3275. Fax: 618 8222 3543. E-mail: heuzenroeder{at}imvs.sa.gov.au.

Journal of Clinical Microbiology, April 2005, p. 1604-1611, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1604-1611.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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