Medycyna doswiadczalna i mikrobiologia · 2013
[Sequence-based typing--molecular typing of Legionella pneumophila strains within the framework of an international external quality assessment].
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Piekarska Katarzyna, Rzeczkowska Magdalena, Stypułkowska-Misiurewicz Hanna
Аннотация
INTRODUCTION: The aim of the study was evaluation results of molecular typing L. pneumophila strains that was carried by using SBT (Sequence-Based Typing) method, obtained by laboratory of Department of Bacteriology NIZP-PZH within the framework of the ninth international external quality assessment (ELDSNet Legionella pneumophila DNA SBT) and their comparision with the results obtained by other reference laboratories in EU.
MATERIAL AND METHODS: The panel of five coded isolates of L. pneumophila was investigated. Genomic DNA of Legionella were extracted and defined regions of seven genes were amplified by PCR and sequenced. Then, consensus sequence of the correct length were generated. In order to determine the complete allelic profile and Seqence Type (ST) forward and reverse sequence for each allele were submitted online by using the L. pneumophila database.
RESULTS: All of L. pneumoniae isolates sent to genotyping by SBT method were correctly identified in our laboratory.
CONCLUSIONS: Results of the ninth international external quality assessment have confirmed competences of laboratory of Department of Bacteriology NIZP-PZH in typing of L. pneumophila isolates accordance with the requirements of the international classification.
MATERIAL AND METHODS: The panel of five coded isolates of L. pneumophila was investigated. Genomic DNA of Legionella were extracted and defined regions of seven genes were amplified by PCR and sequenced. Then, consensus sequence of the correct length were generated. In order to determine the complete allelic profile and Seqence Type (ST) forward and reverse sequence for each allele were submitted online by using the L. pneumophila database.
RESULTS: All of L. pneumoniae isolates sent to genotyping by SBT method were correctly identified in our laboratory.
CONCLUSIONS: Results of the ninth international external quality assessment have confirmed competences of laboratory of Department of Bacteriology NIZP-PZH in typing of L. pneumophila isolates accordance with the requirements of the international classification.