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Journal of Clinical Microbiology, June 1999, p. 1948-1952, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

High-Resolution Genotyping of Streptococcus pyogenes Serotype M1 Isolates by Fluorescent Amplified-Fragment Length Polymorphism Analysis

Meeta Desai,1 Androulla Efstratiou,2 Robert George,2 and John Stanley1,*

Molecular Biology Unit, Virus Reference Division,1 and Streptococcus and Diphtheria Reference Unit, WHO Collaborating Centre for Diphtheria and Streptococcal Infections,2 Central Public Health Laboratory, London NW9 5HT, United Kingdom

Received 11 September 1998/Returned for modification 17 December 1998/Accepted 17 March 1999

We have used fluorescent amplified-fragment length polymorphism (FAFLP) analysis to subtype clinical isolates of Streptococcus pyogenes serotype M1. Established typing methods define most M1 isolates as members of a clone that has a worldwide distribution and that is strongly associated with invasive diseases. FAFLP analysis simultaneously sampled 90 to 120 loci throughout the M1 genome. Its discriminatory power, precision, and reproducibility were compared with those of other molecular typing methods. Irrespective of disease symptomatology or geographic origin, the majority of the clinical M1 isolates shared a single ribotype, pulsed-field gel electrophoresis macrorestriction profile, and emm1 gene sequence. Nonetheless, among these isolates, FAFLP analysis could differentiate 17 distinct profiles, including seven multi-isolate groups. The FAFLP profiles of M1 isolates reproducibly exhibited between 1 and more than 20 amplified fragment differences. The high discriminatory power of genotyping by FAFLP analysis revealed genetic microheterogeneity and differentiated otherwise "identical" M1 isolates as members of a clone complex.

* Corresponding author. Mailing address: Molecular Biology Unit, Virus Reference Division, Central Public Health Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone: 0181 200 4400, ext. 3090. Fax: 0181 200 1569. E-mail: mdesai{at}hgmp.mrc.ac.uk.

Journal of Clinical Microbiology, June 1999, p. 1948-1952, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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