Innovative use of platinum compounds to selectively detect live microorganisms by polymerase chain reaction

Area of Science:

• Microbiology
• Molecular Biology
• Analytical Chemistry

Background:

• Standard PCR cannot differentiate between live and dead microorganisms.
• Existing methods like ethidium/propidium-monoazide (EMA/PMA) are laborious and time-consuming.
• A need exists for a more efficient method to determine microbial viability.

Purpose of the Study:

• To introduce an innovative method using platinum compounds for discriminating live from dead microorganisms.
• To evaluate the efficacy of Platinum-PCR compared to traditional methods like PMA.
• To assess the potential of Platinum-PCR for rapid microbial detection in food and environmental samples.

Main Methods:

• Microorganisms were exposed in vivo to platinum compounds, which selectively bind to DNA in dead cells.
• The Platinum-PCR method was developed to detect viable microorganisms based on platinum compound chelation.
• The method was tested on water and milk samples, including spiked with Cronobacter sakazakii and Escherichia coli.

Main Results:

• Platinum compounds effectively discriminated between live and dead microorganisms in PCR analysis.
• The Platinum-PCR method demonstrated clear differentiation compared to PMA.
• The method enabled specific detection of viable coliforms in milk at low concentrations (5-10 CFU mL(-1)) within 4 hours.

Conclusions:

• Platinum-PCR offers a rapid, efficient, and accurate alternative to traditional methods for determining microbial viability.
• This method has significant potential for application in environmental, food safety, and clinical testing.
• Platinum-PCR could serve as a viable substitute for conventional culture-based methods, improving food safety and public health.