Abstract
The aim of this study was to identify the strengths and weaknesses of five DNA fingerprint methods for epidemiological typing of Streptococcus pneumoniae. We investigated the usefulness of (i) ribotyping, (ii) BOX fingerprinting with the BOX repetitive sequence of S. pneumoniae as a DNA probe, (iii) PCR fingerprinting with a primer homologous to the enterobacterial repetitive intergenic consensus sequence, (iv) pulsed-field gel electrophoresis of large DNA fragments, and (v) restriction fragment end labeling to detect restriction fragment length polymorphism of small DNA fragments. Twenty-eight S. pneumoniae strains isolated from the blood and/or cerebrospinal fluid of 21 patients were analyzed. Genetic clustering among the 28 strains was independent of the DNA fingerprint technique used. However, the discriminatory power and the similarity values differed significantly among the individual techniques. BOX fingerprinting, pulsed-field gel electrophoresis, and restriction fragment end labeling provided the highest degree of discriminatory power. Furthermore, the ease with which computerized fingerprint analysis could be conducted also varied significantly among the techniques. Ribotyping, BOX fingerprinting, and restriction fragment end labeling were very suitable techniques for accurate computerized data analysis. Because of their high discriminatory potential and ease of accurate analysis, we conclude that BOX fingerprinting and restriction fragment end labeling are the most suitable techniques to type pneumococcal strains.
MeSH Terms
Bacterial Typing Techniques
Base Sequence
Consensus Sequence
DNA Fingerprinting/methods
DNA Primers/genetics
DNA Probes/genetics
DNA, Bacterial/blood,cerebrospinal fluid,genetics
Electrophoresis, Gel, Pulsed-Field
Evaluation Studies as Topic
Genotype
Humans
Molecular Epidemiology
Molecular Sequence Data
Pneumococcal Infections/epidemiology,microbiology
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Repetitive Sequences, Nucleic Acid
Streptococcus pneumoniae/classification,genetics,isolation & purification
Chemicals
DNA Primers
DNA Probes
DNA, Bacterial
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hermans P W
Department of Pediatrics, University Hospital Rotterdam, Erasmus University Rotterdam, The Netherlands.
Sluijter M
Hoogenboezem T
Heersma H
van Belkum A
de Groot R
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