Rapid Detection of Antimicrobial Resistance Genes in Critically Ill Children Using a Custom TaqMan Array Card

Author:

Clark John A.12ORCID,Curran Martin D.3,Gouliouris Theodore23,Conway Morris Andrew245ORCID,Bousfield Rachel23,Navapurkar Vilas2,Kean Iain R. L.1ORCID,Daubney Esther1,White Deborah1,Baker Stephen6,Pathan Nazima12

Affiliation:

1. Department of Paediatrics, University of Cambridge, Cambridge CB2 0QQ, UK

2. Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 0QQ, UK

3. Clinical Microbiology and Public Health Laboratory, United Kingdom Health Security Agency, Cambridge CB2 0QQ, UK

4. Division of Anaesthesia, Department of Medicine, University of Cambridge, Cambridge CB2 2QQ, UK

5. Division of Immunology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK

6. Cambridge Institute of Therapeutic Immunology and Infectious Disease, University of Cambridge, Cambridge CB2 0AW, UK

Abstract

Bacteria are identified in only 22% of critically ill children with respiratory infections treated with antimicrobial therapy. Once an organism is isolated, antimicrobial susceptibility results (phenotypic testing) can take another day. A rapid diagnostic test identifying antimicrobial resistance (AMR) genes could help clinicians make earlier, informed antimicrobial decisions. Here we aimed to validate a custom AMR gene TaqMan Array Card (AMR-TAC) for the first time and assess its feasibility as a screening tool in critically ill children. An AMR-TAC was developed using a combination of commercial and bespoke targets capable of detecting 23 AMR genes. This was validated using isolates with known phenotypic resistance. The card was then tested on lower respiratory tract and faecal samples obtained from mechanically ventilated children in a single-centre observational study of respiratory infection. There were 82 children with samples available, with a median age of 1.2 years. Major comorbidity was present in 29 (35%) children. A bacterial respiratory pathogen was identified in 13/82 (16%) of children, of which 4/13 (31%) had phenotypic AMR. One AMR gene was detected in 49/82 (60%), and multiple AMR genes were detected in 14/82 (17%) children. Most AMR gene detections were not associated with the identification of phenotypic AMR. AMR genes are commonly detected in samples collected from mechanically ventilated children with suspected respiratory infections. AMR-TAC may have a role as an adjunct test in selected children in whom there is a high suspicion of antimicrobial treatment failure.

Funder

Addenbrooke’s Charitable Trust, Cambridge University Hospitals

NIHR Cambridge Biomedical Research Centre

Gates Cambridge Trust

Wellcome Trust

Wellcome Trust Clinical Research Career Development Fellowship

MRC Clinician Scientist Fellowship

Action Medical Research

Bill & Melinda Gates Foundation

Publisher

MDPI AG

Subject

Pharmacology (medical),Infectious Diseases,Microbiology (medical),General Pharmacology, Toxicology and Pharmaceutics,Biochemistry,Microbiology

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