Application and evaluation of the interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) for identification of Streptococcus species

• Published in: Journal of clinical microbiology 2001-04, Vol.39 (4), p.1436-1442
• Main Authors: BAELE, Margo, STORMS, Virginie, HAESEBROUCK, Freddy, DEVRIESE, Luc A, GILLIS, Monique, VERSCHRAEGEN, Gerda, DE BAERE, Thierry, VANEECHOUTTE, Mario
• Format: Article
• Language: English
• Subjects: Bacterial diseases; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; capillary electrophoresis; Databases, Factual; DNA Fingerprinting - standards; DNA, Bacterial - genetics; DNA, Intergenic - genetics; Electrophoresis, Capillary - methods; Electrophoresis, Capillary - standards; Fundamental and applied biological sciences. Psychology; Human bacterial diseases; Humans; Infectious diseases; Laboratories - standards; Medical sciences; Microbiology; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - standards; Polymorphism, Genetic - genetics; Reproducibility of Results; RNA, Transfer - genetics; Software; Staphylococcal infections, streptococcal infections, pneumococcal infections; Streptococcal Infections - diagnosis; Streptococcus; Streptococcus - classification; Streptococcus - genetics; tRNA intergenic length polymorphism analysis
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.39.4.1436-1442.2001

The discriminatory power, speed, and interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) combined with capillary electrophoresis was evaluated for the identification of streptococci. This method was carried out in three different laboratories under highly standardized conditions for 54 strains belonging to 18 different species. It was concluded that interlaboratory reproducibility of tDNA fingerprints produced by means of capillary electrophoresis was sufficiently high to permit the exchange between different laboratories and the construction of common libraries which can be consulted for comparison with fingerprints obtained independently in separate laboratories. In a second step, 17 other species were included in the study and examined in one of the participating laboratories. All Streptococcus species studied, except S. mitis, S. oralis, S. parasanguinis, S. pneumoniae, S. thermophilus, and S. vestibularis, showed distinguishable tDNA fingerprints. A database of well-characterized strains was constructed to enable computer-aided identification of unknown streptococcal isolates.