[Sequence-based typing--molecular typing of Legionella pneumophila strains within the framework of an international external quality assessment].

Katarzyna Piekarska, Magdalena Rzeczkowska, Hanna Stypułkowska-Misiurewicz
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Abstract

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.

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[基于序列的分型——在国际外部质量评估框架内嗜肺军团菌菌株的分子分型]。
前言:本研究的目的是对NIZP-PZH细菌学学系实验室在第9次国际外部质量评估(ELDSNet Legionella pneumoniila DNA SBT)框架下采用SBT (Sequence-Based typing)方法进行的嗜肺杆菌分子分型结果进行评价,并与欧盟其他参比实验室的结果进行比较。材料与方法:对5个嗜肺乳杆菌编码分离株进行鉴定。提取军团菌基因组DNA,用PCR扩增7个基因的特定区域并测序。然后,生成正确长度的共识序列。利用嗜肺乳杆菌数据库在线提交每个等位基因的正向和反向序列,以确定完整的等位基因谱和序列型(ST)。结果:本实验室采用SBT法进行基因分型的肺炎乳杆菌分离株均得到正确鉴定。结论:第九次国际外部质量评价结果确认我校细菌学研究室符合国际分类要求,具有开展嗜肺乳杆菌分型工作的能力。
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