Rapid detection of NDM and VIM carbapenemase encoding genes by recombinase polymerase amplification and lateral flow-based detection

Abdulrahman K S Ayfan; Joanne Macdonald; Patrick N A Harris; Claire Heney; David L Paterson; Ella Trembizki; Claire Y T Wang; David M Whiley; Hosam M Zowawi; Adam D Irwin

doi:10.1007/s10096-021-04267-6

Rapid detection of NDM and VIM carbapenemase encoding genes by recombinase polymerase amplification and lateral flow-based detection

Eur J Clin Microbiol Infect Dis. 2021 Nov;40(11):2447-2453. doi: 10.1007/s10096-021-04267-6. Epub 2021 May 11.

Authors

Abdulrahman K S Ayfan^{1

2}, Joanne Macdonald³, Patrick N A Harris^{1

4}, Claire Heney⁴, David L Paterson¹, Ella Trembizki¹, Claire Y T Wang⁵, David M Whiley^{1

4}, Hosam M Zowawi^#^{1

6

7}, Adam D Irwin^#^{8

9}

Affiliations

¹ UQ Centre for Clinical Research, Faculty of Medicine, The University of Queensland, Royal Brisbane and Women's Hospital Campus, Brisbane, Australia.
² Faculty of Science, Biochemistry Department, King Abdul-Aziz University (KAU), Jeddah, Saudi Arabia.
³ School of Science and Engineering, Genecology Research Centre, University of Sunshine Coast (USC), Sunshine Coast, Australia.
⁴ Department of Microbiology, Pathology Queensland, Brisbane, Australia.
⁵ Infection Management and Prevention Service, Queensland Children's Hospital, Brisbane, Australia.
⁶ College of Medicine, King Saud bin Abdul-Aziz University for Health Science (KSAU-HS), Riyadh, Saudi Arabia.
⁷ King Abdullah International Medical Research Centre (KAIMRC), Riyadh, Saudi Arabia.
⁸ UQ Centre for Clinical Research, Faculty of Medicine, The University of Queensland, Royal Brisbane and Women's Hospital Campus, Brisbane, Australia. a.irwin@uq.edu.au.
⁹ Infection Management and Prevention Service, Queensland Children's Hospital, Brisbane, Australia. a.irwin@uq.edu.au.

^# Contributed equally.

PMID: 33974185
DOI: 10.1007/s10096-021-04267-6

Abstract

Carbapenemase-producing organisms (CPOs) pose a serious clinical threat and rapid detection tools are essential to aid in patient management. We developed rapid and simple molecular tests to detect bla_NDM-type and bla_VIM-type carbapenemase genes using recombinase polymerase amplification (RPA) combined with a lateral flow detection. The tests could provide results in approximately 15 min when using DNA extracts, with limits of detection of 9.2 copies/μl for the bla_NDM-type assay and 7.5 copies/μl for bla_VIM-type assay, and successfully detected all isolates harbouring the carbapenemase encoding genes in a panel of 57 isolates. These RPA tests may be suitable for use in low-resource settings to tailor rapid implementation of infection control precautions and antibiotic stewardship.

Keywords: Carbapenemase; Lateral flow; NDM; Point-of-care; Recombinase polymerase amplification; VIM.

Publication types

Evaluation Study

MeSH terms

Anti-Bacterial Agents / pharmacology
Bacteria / drug effects
Bacteria / enzymology*
Bacteria / genetics
Bacterial Proteins / genetics*
Bacterial Proteins / metabolism
DNA Primers / genetics
Drug Resistance, Bacterial
Nucleic Acid Amplification Techniques / instrumentation
Nucleic Acid Amplification Techniques / methods*
Recombinases / metabolism
beta-Lactamases / genetics*
beta-Lactamases / metabolism

Substances

Anti-Bacterial Agents
Bacterial Proteins
DNA Primers
Recombinases
beta-Lactamases
carbapenemase