A modified multilocus sequence typing protocol to genotype Kingella kingae from oropharyngeal swabs without bacterial isolation

BMC Microbiology

Methodology Article

Nawal El Houmami¹  Guillaume André Durand¹  Jean-Christophe Pons¹  Didier Raoult¹  Anis Oubraham¹  Pierre-Edouard Fournier¹  Pablo Yagupsky²  Philippe Minodier³  Dimitri Ceroni⁴  Philippe Bidet⁵  Janek Bzdrenga⁶ 

[1] Aix-Marseille Univ, UM63, CNRS 7278, IRD 198, Inserm 1095, Assistance Publique – Hôpitaux de Marseille, URMITE, Institut Hospitalo-Universitaire Méditerranée Infection, 19-21 Boulevard Jean Moulin, 13385, Marseille, France;Clinical Microbiology Laboratory, Soroka University Medical Center, Beer-Sheva, Israel;Department of Pediatric Emergency Medicine, North Hospital, Aix-Marseille University, Marseille, France;Département de l’enfant et de l’adolescent, HUG-Hôpital des Enfants, Geneva, Switzerland;Laboratoire de Microbiologie, Hôpital Robert Debré, Assistance Publique – Hôpitaux de Paris, Université Paris Diderot, Sorbonne Paris Cité, Inserm, IAME, UMR 1137, Paris, France;University of Grenoble Alpes, CEA, CNRS, IBS, F-38000, Grenoble, France;

关键词: Kingella kingae;    MLST;    Pediatrics;    Outbreaks;    Bone and joint infections;   

DOI  :  10.1186/s12866-017-1104-5

 received in 2017-05-26, accepted in 2017-09-05,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundOutbreaks of Kingella kingae infection are an emerging public health concern among daycare attendees carrying epidemic clones in the oropharynx. However, genotyping of such epidemic clones from affected cases is limited by the low performance of current methods to detect K. kingae from blood samples and lack of specimens available from infected sites. We aimed at developing a modified multilocus sequence typing (MLST) method to genotype K. kingae strains from oropharyngeal samples without prior culture. We designed in silico MLST primers specific for K. kingae by aligning whole nucleotide sequences of abcZ, adk, aroE, cpn60, recA, and gdh/zwf genes from closely related species belonging to the Kingella and Neisseria genera. We tested our modified MLST protocol on all Kingella species and N. meningitidis, as well as 11 oropharyngeal samples from young children with sporadic (n = 10) or epidemic (n = 1) K. kingae infection.ResultsWe detected K. kingae-specific amplicons in the 11 oropharyngeal samples, corresponding to sequence-type 6 (ST-6) in 6 children including the epidemic cases, ST-25 in 2 children, and 3 possible novel STs (ST-67, ST-68, and ST-69). No amplicon was obtained from other Kingella species and N. meningitidis.ConclusionsWe herein developed a specific MLST protocol that enables genotyping of K. kingae by MLST directly from oropharyngeal samples. This discriminatory tool, with which we identified the first K. kingae outbreak caused by ST-6 in Europe, may be used in further epidemiological investigations.

【 授权许可】

CC BY   
© The Author(s). 2017