title: Identification of Legionella spp. by 19 European reference laboratories: results of the European Working Group for Legionella Infections External Quality Assessment Scheme using DNA sequencing of the macrophage infectivity potentiator gene and dedicated online tools.
abstract: Identification of Legionella spp. can be achieved by DNA sequencing of the macrophage infectivity potentiator (mip) gene. The External Quality Assurance (EQA) scheme described in this report is the first to assess the proficiency of laboratories using this methodology. The results obtained from two EQA distributions sent to European reference laboratories involved in Legionella outbreak control and environmental monitoring are presented. Each distribution contained a panel of ten coded Legionella strains. All strains were from clinical and environmental sources and were considered to be wild-type strains. Participants used dedicated online tools to compare sequence text files against a database of known Legionella spp. The majority of centres (seven of ten, and 11 of 12) correctly identified all strains tested, in the first and second distributions, respectively. Typically, sequence similarity values of 98-100% were obtained when the test strains were compared with sequences contained in the database. In all but one case, lower values indicated a poor quality sequence. The exception was associated with the identification of a putative new species in the first panel. Genotypic identification of Legionella can be achieved by the use of standard protocols, dedicated identification libraries, and online tools. EQA schemes provide an independent measure of performance, and it is recommended that laboratories performing these techniques participate in such schemes, thereby allowing optimisation of and improvements in their performance.
authors.affiliation: Respiratory and Systemic Infection Laboratory, Health Protection Agency Centre for Infections, London, UK.
authors.forename: N K
authors.initials: NK
authors.lastname: Fry
authors.forename: B
authors.initials: B
authors.lastname: Afshar
authors.forename: W
authors.initials: W
authors.lastname: Bellamy
authors.forename: A P
authors.initials: AP
authors.lastname: Underwood
authors.forename: R M
authors.initials: RM
authors.lastname: Ratcliff
authors.forename: T G
authors.initials: TG
authors.lastname: Harrison
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D005060:Europe; D006801:Humans; D007875:Legionella / Q000235:genetics* / Q000302:isolation & purification; D019696:Peptidylprolyl Isomerase / Q000235:genetics*; D011786:Quality Control; D012015:Reference Standards; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000592:standards
chemical_list: D001426:Bacterial Proteins; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase