PFGE standard operating procedures for Listeria monocytogenes: harmonizing the typing of food and clinical strains in Europe.
PFGE standard operating procedures for Listeria monocytogenes: harmonizing the typing of food and clinical strains in Europe.
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单核细胞增生李斯特氏菌的 PFGE 标准操作程序：协调欧洲食品和临床菌株的分类。
DOI：
10.1089/fpd.2014.1877
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发表时间：
2015
影响因子：
2.8
通讯作者：
S. Roussel
中科院分区：
文献类型：
--
作者：
D. Michelon;B. Félix;N. Vingadassalon;Jean;J. Larsson;E. Møller;S. Roussel
单核细胞增多性李斯特菌是一种食源性病原体，可导致一种被称为李斯特菌病的严重疾病。欧洲疾病预防和控制中心(ECDC)协调负责临床菌株分型的国家公共卫生实验室(NPHL)网络。在食品方面，它是欧洲联盟单核细胞增多性李斯特菌参考实验室(EURLLM)，管理着一个国家参考实验室网络(NRLS)。自2007年以来，脉冲场凝胶电泳(PFGE)标准操作程序(EURL SOP)已在EURL LM中常规使用。EURL LM建议NRL使用EURL SOP，而根据ECDC合同，统计血清研究所(SSI)要求NHL使用2010年发布的PulseNet USA网络的Halpins SOP(HSOP)。最新的Halpins标准操作规程(UHSOP)于2013年发布。为了促进人类和食品欧洲参考实验室之间的图谱交换，确保用这些不同的SOP获得的PFGE图谱具有可比性是至关重要的。这里的目的是将EURL SOP与HSOP和uHSOP进行比较。该小组由114株特征良好的SSI/EURL菌株组成。所有这些都在EURL上使用EURL SOP和uHSOP进行了表征。在114株菌株中，有70株也在SSI上用HSOP进行了鉴定。EURL、SOP和uHSOP为114株受测菌株产生了难以区分的组合(APAI/ASCI)图谱。EURL、SOP和HSOP为70个测试菌株中的69个产生了难以区分的组合图谱。ASCI图谱显示1株HSOP为184 kBP的低强度条带。对于该菌株，SSI和EUR LM已经从NPHL和NRLS观察到了相同的图谱。然而，这一偏差很小，因为它约占所有114个组合剖面的1%。这项研究应有助于在人类和食品参考实验室之间交换可重现的全氟乙烷凝胶电泳法图谱。
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.