Rapid Detection of Carbapenemase-Producing Enterobacteriacae Based on Surface-Enhanced Raman Spectroscopy with Gold Nanostars

Abstract

The emergence and rapid spread of antibiotic resistance poses a serious threat to healthcare systems across the globe. The existence of carbapenemase-producing Enterobacteriaceae (CPE) such as Klebsiella pneumoniae renders the use of carbapenems, the last-resort class of β-lactam antibiotics, ineffective against bacterial infections, often leading to CPE-associated mortalities. Current methods of detection such as the Carba NP test and modified Hodge's test require hours to days to detect, which delays the response to isolate patients for rapid intervention. Here, we developed a surface-enhanced Raman scattering (SERS)-based detection scheme which utilizes gold nanostars conjugated to a β-lactam antibiotic ceftriaxone (CRO) as a beacon for rapid detection of bacterial β-lactamase secreted by Delhi metalloproteinase (NDM)-producing Escherichia coli as our CPE model with carbapenemase activity. The cleavage of β-lactam ring in CRO by NDM (Class B β-lactamase) caused a detectable reduction in SERS intensities at 722, 1358, and 1495 cm −1 within 25 min. Ratiometric analysis of the SERS peaks at 722, 1358, and 1495 cm −1 normalized against the Raman peak of polystyrene cuvette at 620 cm −1 showed the peak at 1358 cm −1 having the most significant change in intensity upon CPE detection. This reduced detection time has not been reported to date for CPE detection, and our novel approach using SERS could be extended to detect the activity of other classes of β-lactamases to broaden its clinical utility.

Advances in surface‐enhanced Raman spectroscopy technology for detection of foodborne pathogens

“…Wong et al. (2020) used gold nanostar (Au NS) conjugated with the β‐lactam antibiotic ceftriaxone (CRO) as beacon molecule and combined with SERS technology to detect bacterial β‐lactamases secreted by E. coli and, thus, enabled a rapid detection of infections with this class of antibiotic‐resistant bacteria (Figure 6). Rodríguez‐Lorenzo et al.…”

“…I G U R E 6 (A) Schematic diagram for surface-enhanced Raman spectroscopy (SERS) detection of β-lactamase activity with CRO-conjugated Au NS. (B) Characterization of Au NS and Au NS-CRO: (a) UV−vis of AuNS; (b) hydrodynamic size of Au NS; (c) TEM images of AuNS; (d) UV−Vis spectrum of Au NS incubated with varying concentrations of CRO(Wong et al, 2020). Source: Copyright 2020, American Chemical Society.…”

Plasmonic Nanostars: Systematic Review of their Synthesis and Applications

“…In addition to the detection of SARS-CoV-2 − and cancer biomarkers, , many other studies also reported the development of various AuNSt-based biosensors for the detection of bacteria, antibiotics, cortisol, mRNA, or nucleic acids, as summarized in Table . The wide range of biological analytes detected by these AuNSt-enabled sensing platforms is indicative of the advantages and versatility of plasmonic nanostars.…”

Recent advances in antibiotic resistance diagnosis using SERS: focus on the “Big 5” challenges

“…In a different recent study, Wong et al described another single capture probe for the detection of bacterial β-lactamase enzyme secreted by New Delhi metallo-betalactamase (NDM)-producing E. coli . 121 In this study, Au nanostars were conjugated with β-lactam antibiotic ceftriaxone (CRO) and used as a beacon for rapid detection. By incubating the NDM-producing E. coli with the conjugate for 25 min, a detectable reduction in the SERS signal intensity of CRO Raman peaks at 722, 1358, and 1495 cm −1 was noticed due to CRO hydrolysis by NDM-producing E. coli in spite of the fact that the rapid molecular structure change of CRO could be beneficial for early diagnosis and treatment, the method was not able to set a quantifiable limit of detection due to results variations between experiments using a one calibrated loop.…”