Fluorescent amplified-fragment length polymorphism analysis of an outbreak of group A streptococcal invasive disease

Meeta Desai, Asha Tanna, Robert Wall, Androulla Efstratiou, Robert George, John Stanley

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was carried out for an outbreak of group A streptococcal (GAS) invasive disease. Streptococcal genomic DNAs were digested with endonucleases EcoRI and MseI, site-specific adaptors were ligated, and PCR amplification was carried out with an EcoRI adaptor-specific primer labelled with fluorescent dye. Amplified fragments of up to 600 bp in size were separated on a polyacrylamide sequencing gel which contained internal size markers in each lane. These data were automatically scanned and analyzed, fragments were precisely sized (± 1 bp), and electropherograms were generated for each genome with GeneScan 2.1 software. All isolates were compared in this way. Among 27 GAS isolates examined, we found 18 FAFLP profiles, compared with 12 macrorestriction profiles by pulsed-field gel electrophoresis. FAFLP readily distinguished genotypes for two clones of GAS serotype M77 which were responsible for outbreaks of invasive disease in a care-of-the-elderly system. It provided an automated analysis of the whole genome of bacterial isolates. It was reproducible, more discriminatory, and capable of higher throughput than other molecular typing methods. Given agreed conditions, FAFLP would be reproducible between laboratories for rapid characterization of outbreak strains.

Original languageEnglish (US)
Pages (from-to)3133-3137
Number of pages5
JournalJournal of clinical microbiology
Volume36
Issue number11
StatePublished - Nov 1 1998

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Amplified Fragment Length Polymorphism Analysis
Disease Outbreaks
Deoxyribonuclease EcoRI
Bacterial Genomes
Molecular Typing
Pulsed Field Gel Electrophoresis
Fluorescent Dyes
Software
Clone Cells
Genotype
Genome
Polymerase Chain Reaction
DNA

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

Cite this

Desai, Meeta ; Tanna, Asha ; Wall, Robert ; Efstratiou, Androulla ; George, Robert ; Stanley, John. / Fluorescent amplified-fragment length polymorphism analysis of an outbreak of group A streptococcal invasive disease. In: Journal of clinical microbiology. 1998 ; Vol. 36, No. 11. pp. 3133-3137.
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Fluorescent amplified-fragment length polymorphism analysis of an outbreak of group A streptococcal invasive disease. / Desai, Meeta; Tanna, Asha; Wall, Robert; Efstratiou, Androulla; George, Robert; Stanley, John.

In: Journal of clinical microbiology, Vol. 36, No. 11, 01.11.1998, p. 3133-3137.

Research output: Contribution to journalArticle

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