LSPR-Based Biosensing Enables the Detection of Antimicrobial Resistance Genes

Area of Science:

• Biotechnology
• Molecular Biology
• Spectroscopy

Background:

• The demand for rapid, simple, and accurate nucleic acid detection bioassays is increasing.
• Antimicrobial resistance (AMR) poses a significant global health threat, necessitating efficient detection methods.
• The blaSHV gene confers resistance to a broad spectrum of beta-lactam antibiotics.

Purpose of the Study:

• To develop and validate a localized surface plasmon resonance (LSPR) spectroscopy-based bioassay for detecting the blaSHV antimicrobial resistance gene.
• To assess the sensitivity, real-time monitoring capabilities, and specificity of the LSPR system for nucleic acid detection.
• To demonstrate the utility of LSPR in distinguishing between different single nucleotide polymorphism (SNP) variants of the blaSHV gene.

Main Methods:

• Utilized localized surface plasmon resonance (LSPR) spectroscopy for label-free detection of nucleic acids.
• Performed limit of detection experiments to determine the sensitivity for DNA sequences.
• Monitored hybridization events in real-time to extract kinetic parameters and analyzed SNP variants of the blaSHV gene.

Main Results:

• Achieved detection of a 23-nucleotide (nt) deoxyribonucleic acid (DNA) sequence down to 25 nm.
• Demonstrated an inverse correlation between signal intensity and DNA sequence length (23, 43, 63, and 100 nt).
• Successfully distinguished single nucleotide polymorphism (SNP) variants of the blaSHV gene from the fully complementary sequence using LSPR.

Conclusions:

• The developed LSPR system provides a robust, label-free, and cost-efficient analytical tool for nucleic acid detection.
• The system enables real-time monitoring and kinetic analysis of hybridization events.
• This technology is crucial for the surveillance of antimicrobial resistance determinants and can aid in clinical decision-making for antibiotic treatment.