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Journal of Clinical Microbiology, September 2002, p. 3406-3415, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3406-3415.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Fluorescent Amplified Fragment Length Polymorphism Analysis of Salmonella enterica Serovar Typhimurium Reveals Phage-Type- Specific Markers and Potential for Microarray Typing

Honghua Hu, Ruiting Lan,^, and Peter R. Reeves^*

School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, New South Wales 2006, Australia

Received 26 December 2001/ Returned for modification 9 March 2002/ Accepted 17 June 2002

Fluorescent amplified fragment length polymorphism (AFLP) was applied to 46 Salmonella enterica serovar Typhimurium isolates of Australian origin comprising nine phage types, by using the restriction enzymes MseI and EcoRI and all 16 possible MseI +1-EcoRI +1 primer pair combinations. AFLP in the present study showed a very good discrimination power with a Simpson index of diversity of 0.98, and 35 different AFLP patterns were observed in the 46 isolates. AFLP grouped most serovar Typhimurium isolates by phage type and enabled differentiation of phage types. Furthermore, 84 phage-type-specific polymorphic AFLP fragments, for which presence or absence correlated with phage type (including 25 with one exception to phage type specificity) were observed in the 46 strains studied. Eighteen phage-type-specific AFLP fragments were cloned and sequenced. Fifteen are of known genes or have a homologue in the databases. Three sequences are plasmid related, eight are phage related, and four relate to chromosomal genes. Twelve of the 18 fragments are polymorphic because the DNA is present or absent as indicated by Southern hybridization, and we see good potential to use sequences of these fragments as the basis for multiplex PCR and development of a microarray-based molecular phage-typing method for serovar Typhimurium.

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* Corresponding author. Mailing address: School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, New South Wales 2006, Australia. Phone: (612) 9351 2536. Fax: (612) 9351 4571. E-mail: reeves{at}angis.usyd.edu.au.

Present address: School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, New South Wales 2052, Australia.

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Journal of Clinical Microbiology, September 2002, p. 3406-3415, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3406-3415.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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