Semi-automatic instrumentation for nucleic acid extraction and purification to quantify pathogens on surfaces

Area of Science:

• Microbiology
• Environmental Health
• Biotechnology

Background:

• Public lavatories are high-traffic areas that can facilitate the spread of infectious pathogens.
• Surface microbial contamination in public restrooms serves as a key indicator of hygiene control effectiveness.
• Existing methods for pathogen detection may lack sensitivity or speed for effective hygiene monitoring.

Purpose of the Study:

• To develop and validate a novel, sensitive, and rapid method for detecting microbial contamination on surfaces in public lavatories.
• To enhance the detection sensitivity and reproducibility of pathogen analysis using polymerase chain reaction (PCR)-compatible tools.
• To introduce a semi-automatic system for efficient surface sampling and pathogen preparation.

Main Methods:

• Development of PCR-compatible surface sample preparation tools utilizing half A4-sized wipes for pathogen collection.
• A four-step cartridge process: pathogen transfer to aqua phase, concentration via graphene adsorption, lysis using bead-beating, and nucleic acid collection.
• Amplification of collected DNA/RNA using polymerase chain reaction (PCR) or loop-mediated isothermal amplification (LAMP).

Main Results:

• The developed wipe-based method achieved at least one order of magnitude higher sensitivity compared to conventional swab methods.
• The semi-automatic cartridge system demonstrated effective and reproducible pathogen detection in a lavatory hygiene test.
• The method significantly increased detection sensitivity and reproducibility within a short timeframe.

Conclusions:

• The novel wipe sampling method combined with a semi-automatic cartridge offers a highly sensitive and rapid approach for assessing microbial contamination in public lavatories.
• This technology provides a valuable tool for improving hygiene control and public health surveillance in shared environments.
• The enhanced sensitivity and efficiency of this method represent a significant advancement in environmental microbiology and hygiene monitoring.