Current and Future Technologies for the Detection of Antibiotic-Resistant Bacteria

Abstract

Antibiotic resistance is a global public health concern, posing a significant threat to the effectiveness of antibiotics in treating bacterial infections. The accurate and timely detection of antibiotic-resistant bacteria is crucial for implementing appropriate treatment strategies and preventing the spread of resistant strains. This manuscript provides an overview of the current and emerging technologies used for the detection of antibiotic-resistant bacteria. We discuss traditional culture-based methods, molecular techniques, and innovative approaches, highlighting their advantages, limitations, and potential future applications. By understanding the strengths and limitations of these technologies, researchers and healthcare professionals can make informed decisions in combating antibiotic resistance and improving patient outcomes.

Keywords: PCR; antibiotic-resistant bacteria; antimicrobial resistance (AMR); aptamers; biosensors.

Figure 1

(a) An example describes a colorimetric sensor that detects red-to-purple transitions using gold nanoparticles (AuNPs). The left side of the diagram shows the first phase, where the aptamer coating evenly distributes AuNPs, turning them red. AuNPs aggregated after target–aptamer binding and salt addition. (b) An example of aptamer enzyme-linked sorbent assays is shown. The main antibody targets the molecule, while the secondary antibody can be used in different sandwich testing. (a,b) Visible color changes that may be measured with optical equipment.

Figure 2

CRISPR-Cas-based diagnostics. (I) A logic gate type “AND” of CRISPR-Cas12/13-based diagnostics. Amplified nucleic acid from patient samples can be used as input. Attomolar RNA concentrations in serum or urine samples could be detected. Without purification, Cas13 could detect RNA in samples with serum concentration up to 2%. The diagnostic can use cfDNA liquid biopsy samples and DNA extracted from anal swabs. The diagnostic process also requires the Cas12/Cas13 system and a crRNA configuration that matches a target gene, such as an antibiotic resistance gene. The Cas12/Cas13-crRNA combination produces fluorescence output when it identifies target-positive material. The fluorescent signal is a result of a collateral trans-cleavage of the quenched-fluorescent ssDNA/ssRNA by Cas12/Cas13, respectively. (II) CRISPR-Cas9 enrichment for NGS detection. (a) DNA or cDNA sequences harboring the gene of interest are present (green). (b) DNA extremities are obstructed. (c) CRISPR-Cas9 system cleaves the target gene into fragments suitable for NGS (yellow arrows). (d) Ligation of universal sequencing adapters. (e) The target sequence is enriched, which is ready for sequencing.