Biopolymeric surface modified 96-well plates for equipment-free high-throughput DNA extraction and direct ACK-LAMP colorimetric detection of foodborne pathogen.

Ensuring the safety and high quality of food relies on promptly and clearly identifying foodborne diseases, including Salmonella . Nucleic acid testing plays an important role in ensuring food safety. Commercially available high-throughput molecular extraction technologies in-field application is limited as they are reliant on specialized and expensive equipment. The 96-well microplate recycling through biopolymer inner surface coating, was confirmed with FTIR, XRD, and XPS. Once optimized the DNA extraction protocol was equipment-free, user and eco-friendly while reducing sample contamination. Using the biopolymer microplate, the extraction and direct LAMP amplification of Salmonella spp. from milk samples were assessed. The gDNA had an average 260/280 ratio of 1.97, with a DNA yield of 16.88 ± 5.40 μg·mL -1 from 9 × 10 10 CFU·mL -1 Salmonella . Subsequently, using acid chrome K we visually detected the presence of Salmonella . The quantity of bacteria was correlated to the increase in the blue intensity of the color indicator. A detection limit of 1.54 CFU·mL -1 was achieved with good specificity and sensitivity in an equipment free setting. This study successfully established a 96-well plate-based high-throughput equipment free DNA extraction method. The methods enable the high-throughput extraction and amplification of 96 samples concomitantly within 1hr 30 min corresponding respectively to a total of 1536 and 768 samples within a working day treated by a researcher without the use of specialized and expensive equipment. The polymeric modified microplate was used for high-throughput DNA extraction and direct molecular amplification for pathogen detection. High-throughput DNA extraction no longer requires cumbersome equipment.