title: PFGE standard operating procedures for Listeria monocytogenes: harmonizing the typing of food and clinical strains in Europe.
abstract: Listeria monocytogenes is a foodborne pathogen responsible for a severe disease known as listeriosis. The European Centre for Disease Prevention and Control (ECDC) coordinates a network of national public health laboratories (NPHLs) in charge of typing clinical strains. In food, it is the European Union Reference Laboratory for L. monocytogenes (EURL Lm), which manages a network of National Reference Laboratories (NRLs). A pulsed-field gel electrophoresis (PFGE) standard operating procedure (EURL SOP) has been used routinely at the EURL Lm since 2007. The EURL Lm has recommended that NRLs use the EURL SOP, whereas the Statens Serum Institut (SSI), under contract for ECDC, requested that NPHLs use Halpins' SOP (HSOP) published in 2010 for the PulseNet USA network. An update of Halpins' SOP (uHSOP) was published in 2013. To facilitate the exchange of profiles among human and food European reference laboratories, it is crucial to ensure that the PFGE profiles obtained with these different SOPs are comparable. The aim here was to compare the EURL SOP with HSOP and uHSOP. The panel comprised 114 well-characterized SSI/EURL strains. All were characterized at the EURL using both the EURL SOP and uHSOP. Seventy of the 114 strains were also characterized at the SSI using HSOP. The EURL SOP and uHSOP produced indistinguishable combined (ApaI/AscI) profiles for the 114 strains tested. The EURL SOP and HSOP produced indistinguishable combined profiles for 69 of the 70 strains tested. One strain displayed for the AscI profile an additional low-intensity band at 184 kbp with HSOP. For this strain, SSI and EUR Lm had already observed the same profile from NPHLs and NRLs. However, this deviation is minor as it accounted for about 1% of all the 114 combined profiles. This study should facilitate the exchange of reproducible PFGE profiles among human and food reference laboratories.
authors: 1 University Paris-Est , French Agency for Food, Environmental, and Occupational Health & Safety (ANSES), Laboratory for Food Safety, European Union Reference Laboratory for Listeria monocytogenes, Maisons-Alfort, France .
authors: Damien
authors: D
authors: Michelon
authors: Benjamin
authors: B
authors: Félix
authors: Noemie
authors: N
authors: Vingadassalon
authors: Jean-François
authors: JF
authors: Mariet
authors: Jonas T
authors: JT
authors: Larsson
authors: Eva
authors: E
authors: Møller-Nielsen
authors: Sophie
authors: S
authors: Roussel
mesh_terms: D015373:Bacterial Typing Techniques; D016521:Electrophoresis, Gel, Pulsed-Field / Q000592:standards*; D005060:Europe; D005062:European Union; D005516:Food Microbiology; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification* / Q000302:isolation & purification*; D008088:Listeriosis / Q000453:epidemiology / Q000382:microbiology*; D012703:Serotyping