Foodborne pathogenic bacteria have been considered as a major risk factor for food safety. It is of great significance to carry out in-field screening of pathogenic bacteria to prevent the outbreaks of foodborne disea...Foodborne pathogenic bacteria have been considered as a major risk factor for food safety. It is of great significance to carry out in-field screening of pathogenic bacteria to prevent the outbreaks of foodborne diseases. In this study, a portable lab-on-a-disc platform with a microfluidic disc was developed for rapid and automatic detection of Salmonella typhimurium using a nickel nanowire(Ni NW) net for effective separation of target bacteria, horseradish peroxidase nanoflowers(HRP NFs) for efficient amplification of biological signals, and a self-developed smartphone APP for accurate analysis of colorimetric images. First,the microfluidic disc was preloaded with reagents and samples and centrifuged to form one bacterial sample column, one immune Ni NW column, one HRP NF column, two washing buffer columns and one tetramethylbenzidine(TMB) column, which were separated by air gaps. Then, a rotatable magnetic field was specifically developed to assemble the Ni NWs into a net, which was automatically controlled by a stepped motor to successively pass through the sample column for specific capture of target bacteria, the HRP NF column for specific label of target bacteria, the washing columns for effective removal of sample background and non-specific binding NFs, and the TMB column for colorimetric determination of target bacteria. The color change of TMB from colorless to blue was finally analyzed using the smartphone APP to quantitatively determine the target bacteria. This lab-on-a-disc platform could detect Salmonella typhimurium from 5.6 × 10^(1) CFU/20 μL to 5.6 × 10^(5) CFU/20 μL in 1 h with a lower detection limit of 56 CFU/20 μL. The recovery of target bacteria in spiked chicken samples ranged from 97.5% to 101.8%. This portable platform integrating separation, labeling, washing, catalysis and detection onto a single disc is featured with automatic operation, fast reaction, and small size and has shown its potential for in-field detection of foodborne pathogens.展开更多
基金supported by National Natural Science Foundation of China (No. 32071899)Walmart Foundation (No.61626817)Walmart Food Safety Collaboration Center for its great support。
文摘Foodborne pathogenic bacteria have been considered as a major risk factor for food safety. It is of great significance to carry out in-field screening of pathogenic bacteria to prevent the outbreaks of foodborne diseases. In this study, a portable lab-on-a-disc platform with a microfluidic disc was developed for rapid and automatic detection of Salmonella typhimurium using a nickel nanowire(Ni NW) net for effective separation of target bacteria, horseradish peroxidase nanoflowers(HRP NFs) for efficient amplification of biological signals, and a self-developed smartphone APP for accurate analysis of colorimetric images. First,the microfluidic disc was preloaded with reagents and samples and centrifuged to form one bacterial sample column, one immune Ni NW column, one HRP NF column, two washing buffer columns and one tetramethylbenzidine(TMB) column, which were separated by air gaps. Then, a rotatable magnetic field was specifically developed to assemble the Ni NWs into a net, which was automatically controlled by a stepped motor to successively pass through the sample column for specific capture of target bacteria, the HRP NF column for specific label of target bacteria, the washing columns for effective removal of sample background and non-specific binding NFs, and the TMB column for colorimetric determination of target bacteria. The color change of TMB from colorless to blue was finally analyzed using the smartphone APP to quantitatively determine the target bacteria. This lab-on-a-disc platform could detect Salmonella typhimurium from 5.6 × 10^(1) CFU/20 μL to 5.6 × 10^(5) CFU/20 μL in 1 h with a lower detection limit of 56 CFU/20 μL. The recovery of target bacteria in spiked chicken samples ranged from 97.5% to 101.8%. This portable platform integrating separation, labeling, washing, catalysis and detection onto a single disc is featured with automatic operation, fast reaction, and small size and has shown its potential for in-field detection of foodborne pathogens.
