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A modified multilocus sequence typing protocol to genotype Kingella kingae from oropharyngeal swabs without bacterial isolation.
- Source :
-
BMC microbiology [BMC Microbiol] 2017 Sep 21; Vol. 17 (1), pp. 200. Date of Electronic Publication: 2017 Sep 21. - Publication Year :
- 2017
-
Abstract
- Background: Outbreaks 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.<br />Results: We 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.<br />Conclusions: We 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.
- Subjects :
- Bacterial Proteins genetics
Child
Disease Outbreaks
Europe
Humans
Kingella kingae classification
Molecular Epidemiology
Neisseria genetics
Neisseriaceae Infections diagnosis
Neisseriaceae Infections epidemiology
Neisseriaceae Infections microbiology
Polymerase Chain Reaction methods
Species Specificity
Genotype
Genotyping Techniques methods
Kingella kingae genetics
Kingella kingae isolation & purification
Multilocus Sequence Typing methods
Oropharynx microbiology
Subjects
Details
- Language :
- English
- ISSN :
- 1471-2180
- Volume :
- 17
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- BMC microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 28946848
- Full Text :
- https://doi.org/10.1186/s12866-017-1104-5