Multiplexed Ultrafast Photothermal Digital Polymerase Chain Reaction Based on Oligo(Phenylene-Ethynylene)

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Microbiology

Background:

• Periodontal disease diagnosis is currently limited by lengthy, costly, and time-consuming methods.
• An imbalance between oral bacteria and the host immune system underlies periodontal disease.
• Existing diagnostic techniques lack the speed and sensitivity required for timely intervention.

Purpose of the Study:

• To develop an ultrafast, highly sensitive, and multitargeted diagnostic method for periodontal disease.
• To utilize a novel plasmonic photothermal-based digital PCR (dPCR) chip for enhanced detection capabilities.
• To enable real-time, point-of-care molecular diagnostics for periodontal infections.

Main Methods:

• Development of a novel plasmonic photothermal-based dPCR chip incorporating photothermal materials and organic interfacial chemicals.
• Application of ultrafast plasmonic photothermal-based dPCR for amplifying multiple target genes (Streptococcus mutans, Porphyromonas gingivalis, Campylobacter rectus, Prevotella nigrescens) in under 14 minutes.
• Utilizing a fluorescence scanning system for rapid fluorescence intensity measurement (within 9 minutes).

Main Results:

• Achieved amplification of multiple target genes in 14 minutes and fluorescence detection in 9 minutes.
• Demonstrated high sensitivity of 10^1 copies/μL for detecting standard bacteria and bacteria in artificial saliva.
• Successfully performed multiplex PCR on bacterial mixtures containing four species, confirming platform's multitargeting capability.

Conclusions:

• The developed plasmonic photothermal-based dPCR platform offers ultrafast and highly sensitive detection of periodontal pathogens.
• This technology has the potential to revolutionize periodontal disease diagnosis, enabling real-time, point-of-care molecular testing.
• The platform's versatility supports application in various diagnostic fields beyond periodontal disease.