Validation of a novel molecular diagnostic panel for pediatric musculoskeletal infections: Integration of the Cepheid Xpert MRSA/SA SSTI and laboratory-developed real-time PCR assays for clindamycin resistance genes and Kingella kingae detection.


Journal

Journal of microbiological methods
ISSN: 1872-8359
Titre abrégé: J Microbiol Methods
Pays: Netherlands
ID NLM: 8306883

Informations de publication

Date de publication:
01 2019
Historique:
received: 12 04 2018
revised: 06 12 2018
accepted: 07 12 2018
pubmed: 12 12 2018
medline: 1 2 2020
entrez: 12 12 2018
Statut: ppublish

Résumé

Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care. Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a "Musculoskeletal Diagnostic Panel" (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results. A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%. This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections.

Sections du résumé

BACKGROUND
Pathogen detection in pediatric patients with musculoskeletal infections relies on conventional bacterial culture, which is slow and can delay antimicrobial optimization. The ability to rapidly identify causative agents and antimicrobial resistance genes in these infections may improve clinical care.
METHODS
Convenience specimens from bone and joint samples submitted for culture to Children's Hospital Colorado (CHCO) from June 2012 to October 2016 were evaluated using a "Musculoskeletal Diagnostic Panel" (MDP) consisting of the Xpert MRSA/SA SSTI real-time PCR (qPCR, Cepheid) and laboratory-developed qPCRs for Kingella kingae detection and erm genes A, B, and C which confer clindamycin resistance. Results from the MDP were compared to culture and antimicrobial susceptibility testing (AST) results.
RESULTS
A total of 184 source specimens from 125 patients were tested. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Xpert MRSA/SA SSTI compared to culture and AST results were 85%, 98%, 93%, and 95% respectively for MSSA and 82%, 100%, 100%, and 99% for MRSA. Compared to phenotypic clindamycin resistance in S. aureus isolates, the erm A, B, and C gene PCRs collectively demonstrated a sensitivity, specificity, PPV, and NPV of 80%, 96%, 67%, and 98%. In comparison to clinical truth, Kingella PCR had a sensitivity, specificity, PPV, and NPV of 100%, 99.5%, 100%, and 100%.
CONCLUSIONS
This novel MDP offers a rapid, sensitive, and specific option for pathogen detection in pediatric patients with musculoskeletal infections.

Identifiants

pubmed: 30527965
pii: S0167-7012(18)30290-2
doi: 10.1016/j.mimet.2018.12.004
pii:
doi:

Substances chimiques

Anti-Bacterial Agents 0
Bacterial Proteins 0
Clindamycin 3U02EL437C
Methyltransferases EC 2.1.1.-
ErmA protein, Bacteria EC 2.1.1.184
rRNA (adenosine-O-2'-)methyltransferase EC 2.1.1.230

Types de publication

Journal Article Research Support, Non-U.S. Gov't Validation Study

Langues

eng

Sous-ensembles de citation

IM

Pagination

60-67

Informations de copyright

Copyright © 2018 Elsevier B.V. All rights reserved.

Auteurs

Justin B Searns (JB)

Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Christine C Robinson (CC)

Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Qi Wei (Q)

Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Ji Yuan (J)

Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Stacey Hamilton (S)

Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Kristin Pretty (K)

Microbiology Department, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Nathan Donaldson (N)

Department of Orthopedic Surgery, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Sarah K Parker (SK)

Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA.

Samuel R Dominguez (SR)

Division of Pediatric Infectious Diseases, Children's Hospital Colorado, 13123 East 16th Avenue, Aurora, CO 80045, USA. Electronic address: Samuel.dominguez@childrenscolorado.org.

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Classifications MeSH