| Title: |
KpSC-ID: a multiplex real-time PCR assay for the simultaneous detection of the Klebsiella pneumoniae species complex and specific identification of Klebsiella pneumoniae, Klebsiella quasipneumoniae and Klebsiella variicola |
| Authors: |
Mcandrew, Grainne; Barbier, Elodie; Rodrigues, Carla; Piveteau, Pascal; Brisse, Sylvain; Reddington, Kate |
| Contributors: |
School of Biological and Chemical Sciences Galway, Irlande; National University of Ireland Galway (NUI Galway); Agroécologie Dijon; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Agro Dijon; Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Université Bourgogne Europe (UBE); Biodiversité et Epidémiologie des Bactéries pathogènes - Biodiversity and Epidemiology of Bacterial Pathogens; Institut Pasteur Paris (IP)-Université Paris Cité (UPCité); Centre national de Référence de la Coqueluche et autres Bordetelloses - National Reference Center for Whooping Cough and other Bordetella infections (CNR); European Reference laboratory for Public Health on Diphtheria and Pertussis (EURL-PH-DIPE); European Centre for Disease Prevention and Control Solna, Sweden (ECDC); Union Européenne = European Union (UE)-Union Européenne = European Union (UE); Optimisation des procédés en Agriculture, Agroalimentaire et Environnement (UR OPAALE); Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE); This work was supported financially by the MedVetKlebs project from the European Joint Programme One Health, which has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement no. 773830.; European Project: 773830,H2020-SFS-2017-1,MedVetKlebs (a component of European Joint Programme One Health)(2018) |
| Source: |
ISSN: 1350-0872. |
| Publisher Information: |
CCSD; Microbiology Society |
| Publication Year: |
2025 |
| Subject Terms: |
Klebsiella pneumoniae species complex; identification; differentiation; [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology |
| Description: |
International audience ; The Klebsiella pneumoniae species complex (KpSC) comprises five closely related bacterial species, namely Klebsiella pneumoniae, Klebsiella quasipneumoniae, Klebsiella variicola, Klebsiella quasivariicola and Klebsiella africana. The KpSC is ubiquitous in the environment and is also an important human pathogen, particularly associated with healthcare-associated infections. The accurate detection and differentiation of the KpSC is challenging owing to the close phenotypic and genotypic identity (93-95% average nucleotide identity) shared between these members. Current diagnostic assays either fail to detect and identify all KpSC members or misidentify some KpSC members as K. pneumoniae sensu stricto. It is currently estimated that similar to 20% of human infections are caused by members of the KpSC other than K. pneumoniae. This leads to underreporting of some KpSC members in both clinical and environmental settings, which impacts our understanding of the importance of each species. Furthermore, it limits our understanding of the global and local epidemiological impact of some members of the KpSC. In this study, a rapid multiplex real-time PCR assay (KpSC-ID) was designed and developed to detect all KpSC members while simultaneously identifying the predominant human pathogens K. pneumoniae, K. quasipneumoniae and K. variicola. Assay performance was verified in silico using a panel of over 1,000 publicly available genome sequences and experimentally validated using a panel of genomic DNA extracted from 54 Enterobacteriaceae. The assay displayed excellent specificity against over 1,000 genome sequences tested in silico. During in vitro validation, the pan-KpSC assay detected each (29/29) KpSC species and strains tested. For the species-specific assays, 100% specificity was demonstrated in the K. pneumoniae, K. quasipneumoniae and K. variicola assays, respectively. Sensitivity of 10 genomic equivalents was demonstrated for each assay. Ultimately, the diagnostic assay developed in this study ... |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| Relation: |
info:eu-repo/grantAgreement//773830/EU/Klebsiella pneumoniae: from ecology to source attribution and transmission control/MedVetKlebs (a component of European Joint Programme One Health); WOS: 001542230700001 |
| DOI: |
10.1099/mic.0.001587 |
| Availability: |
https://hal.inrae.fr/hal-05489562; https://hal.inrae.fr/hal-05489562v1/document; https://hal.inrae.fr/hal-05489562v1/file/mic001587.pdf; https://doi.org/10.1099/mic.0.001587 |
| Rights: |
https://creativecommons.org/licenses/by/4.0/ ; info:eu-repo/semantics/OpenAccess |
| Accession Number: |
edsbas.C86CF44B |
| Database: |
BASE |