Activity fingerprinting of AMR β-lactamase towards a fast and accurate diagnosis

Front Cell Infect Microbiol. 2023 Sep 5:13:1222156. doi: 10.3389/fcimb.2023.1222156. eCollection 2023.

Abstract

Antibiotic resistance has become a serious threat to global public health and economic development. Rapid and accurate identification of a patient status for antimicrobial resistance (AMR) are urgently needed in clinical diagnosis. Here we describe the development of an assay method for activity fingerprinting of AMR β-lactamases using panels of 7 β-lactam antibiotics in 35 min. New Deli Metallo β-lactamase-1 (NDM-1) and penicillinase were demonstrated as two different classes of β-lactamases. The panel consisted of three classes of antibiotics, including: penicillins (penicillin G, piperacillin), cephalosporins (cefepime, ceftriaxone, cefazolin) and carbapenems (meropenem and imipenem). The assay employed a scheme combines the catalytic reaction of AMR β-lactamases on antibiotic substrates with a flow-injected thermometric biosensor that allows the direct detection of the heat generated from the enzymatic catalysis, and eliminates the need for custom substrates and multiple detection schemes. In order to differentiate classes of β-lactamases, characterization of the enzyme activity under different catalytic condition, such as, buffer composition, ion strength and pH were investigated. This assay could provide a tool for fast diagnosis of patient AMR status which makes possible for the future accurate treatment with selected antibiotics.

Keywords: NDM-1; antibiotics; antimicrobial resistance; thermometric biosensor; β-lactamase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents*
  • Carbapenems / pharmacology
  • Cefazolin
  • Cefepime
  • Humans
  • beta-Lactamases*

Substances

  • beta-Lactamases
  • Anti-Bacterial Agents
  • Carbapenems
  • Cefazolin
  • Cefepime

Grants and funding

This work was supported by Swedish Research council [VR 5607,2014], Swedish Foundation for Strategic Research [RBP14-0055], the four “batches” Innovation Project of Invigorating Medical Through Science and Technology of Shanxi Province (No. 2022XM13), the Programme of Introducing Talents of Discipline to Universities (No. D21004), National Key Research and Development Project of China (No. 2019YFF0216504), the Innovation Program of graduate in Shanxi (No. SXU2022Y220).