Rapid Identification and Classification of Pathogens That Produce Carbapenemases and Cephalosporinases with a Colorimetric Paper-Based Multisensor

Abstract

Infections caused by bacteria that produce β-lactamases (BLs) are a major problem in hospital settings. The phenotypic detection of these bacterial strains requires culturing samples prior to analysis. This procedure may take up to 72 h, and therefore it cannot be used to guide the administration of the first antibiotic regimen. Here, we propose a multisensor for identifying pathogens bearing different types of β-lactamases above the infectious dose threshold within 90 min that does not require culturing samples. Instead, bacterial cells are preconcentrated in the cellulose scaffold of a paper-based multisensor. Then, 12 assays are performed in parallel to identify whether the pathogens produce carbapenemases and/or cephalosporinases, including metallo-β-lactamases, extended-spectrum β-lactamases (ESBLs), and AmpC enzymes. The multisensor generates an array of colored spots that can be quantified with image processing software and whose interpretation leads to the detection of the different enzymes depending on their specificity toward the hydrolysis of certain antibiotics, and/or their pattern of inhibition or cofactor activation. The test was validated for the diagnosis of urinary tract infections. The inexpensive paper platform along with the uncomplicated colorimetric readout makes the proposed prototypes promising for developing fully automated platforms for streamlined clinical diagnosis.

Pretreatment-Free Dual-Excitation Three-Mode Detection of β-Lactamase in Milk and Bacteria by Cr3+-Doped ZnAl2O4 Phosphors

“…Antibiotics have been widely used for the therapy of various diseases of bacterial infection, − but the overuse of antibiotics has brought the negative effect of antibiotic-resistant bacteria. − The most common mechanism of antibiotic resistance of bacteria is to produce β-lactamase, ,− which cleaves the β-lactam ring and thus inactivates the antibacterial activity of the antibiotics. , In addition, β-lactamase is illegally added to contaminated milk to cover up β-lactam antibiotic residues. − Consequently, the rapid and accurate detection of β-lactamase is of paramount significance for customized treatments and food safety.…”

Culture-Independent Multiplexed Detection of Drug-Resistant Bacteria Using Surface-Enhanced Raman Scattering

“…Biochemical assays, such as the Carba NP test, can detect in vitro hydrolysis of the β-lactam ring of imipenem by carbapenemases. , Lateral flow immune assays (LFIAs) have been developed to directly detect resistance protein markers, including various types of β-lactamases . Substrate-based enzyme function assays, such as colorimetric arrays and ESBL-specific fluorogenic probes, have recently gained attention for detecting lactamase-producing bacteria. − Upon activation by β-lactamase, these probes release the fluorescent product that can be recorded on a microscope. The resulting fluorescence can be related to the activities of ESBL-E.…”