Multiplex detection of bacterial pathogens by PCR/SERS assay

Area of Science:

• Biomedical Diagnostics
• Molecular Biology
• Nanotechnology

Background:

• Bacterial infections are a major global health concern, necessitating accurate and rapid diagnostic tools.
• Current PCR-based methods for multiplex pathogen detection face limitations in sensitivity and cost-effectiveness.
• There is a critical need for advanced diagnostic approaches to overcome existing challenges in identifying multiple bacterial pathogens simultaneously.

Purpose of the Study:

• To develop and validate a simple multiplex PCR/SERS assay for the simultaneous detection of four key bacterial pathogens.
• To enhance the sensitivity and specificity of bacterial pathogen identification compared to existing methods.
• To provide a potentially cost-effective and efficient diagnostic tool for clinical applications.

Main Methods:

• A multiplex PCR assay was designed using specific primers for amplifying genomic DNA (gDNA) of *P. aeruginosa*, *S. aureus*, *S. epidermidis*, and *M. smegmatis*.
• Surface-enhanced Raman spectroscopy (SERS) nanotags functionalized with complementary DNA probes and Raman reporters were employed for pathogen identification.
• The assay combined PCR amplification with SERS detection for simultaneous genotyping of bacterial pathogens.

Main Results:

• The developed PCR/SERS assay exhibited high specificity and sensitivity in detecting bacterial pathogen gDNA.
• The assay successfully identified as few as 100 copies of target gDNA, demonstrating significant sensitivity.
• Simultaneous detection of four distinct bacterial pathogens was achieved with the proposed method.

Conclusions:

• The proposed multiplex PCR/SERS assay offers a sensitive and specific method for simultaneous bacterial pathogen detection.
• This assay leverages standard PCR amplification, making it potentially adaptable for clinical settings.
• The developed approach holds significant promise for improving the diagnosis of multiple bacterial infections and aiding clinical decision-making.