Integrated microfluidic systems with sample preparation and nucleic acid amplification

Rapid, efficient and accurate nucleic acid molecule detection is important in the screening of diseases and pathogens, yet remains a limiting factor at point of care (POC) treatment. Microfluidic systems are characterized by fast, integrated, miniaturized features which provide an effective platform for qualitative and quantitative detection of nucleic acid molecules. The nucleic acid detection process mainly includes sample preparation and target molecule amplification. Given the advancements in theoretical research and technological innovations to date, nucleic acid extraction and amplification integrated with microfluidic systems has advanced rapidly. The primary goal of this review is to outline current approaches used for nucleic acid detection in the context of microfluidic systems. The secondary goal is to identify new approaches that will help shape future trends at the intersection of nucleic acid detection and microfluidics, particularly with regard to increasing disease and pathogen detection for improved diagnosis and treatment

A Specific and Sensitive Aptamer-Based Digital PCR Chip for Salmonella typhimurium Detection

Food poisoning and infectious diseases caused by Salmonella typhimurium (S. typhimurium) are serious public health concerns for human health and food safety. The diversity and complexity of food matrices pose great challenges for rapid and ultra-sensitive detection of S. typhimurium in food samples. A method capable of identification, detection, and quantification of S. typhimurium is essential for addressing these issues. In this study, aptamer-coated magnetic beads (Apt-MBs) are employed as capture bio-probes to specifically and selectively concentrate S. typhimurium in food samples. A self-priming chip-based digital PCR was then presented as another biosensor for on-site detection and quantification of S. typhimurium cells. The chip we developed was robust and did not require any external power for sample loading. The combination of Apt-MBs with an on-chip digital detection realized the integration into lab-on-a-chip-based biosensors for on-site monitoring of foodborne pathogens. It was possible to capture and detect S. typhimurium cells as low as 90 CFU/reaction with a capture efficiency of 94.5%. Additionally, the whole process only took about 2 h. This unique platform could also be used to monitor other target bacteria with high specificity and sensitivity by utilizing different aptamers. Furthermore, the platform has potential applications in point-of-care testing in the future

Joint Intra/Inter-Slot Code Design for Unsourced Multiple Access in 6G Internet of Things

Unsourced multiple access (UMA) is the technology for massive, low-power, and uncoordinated Internet-of-Things in the 6G wireless system, improving connectivity and energy efficiency on guaranteed reliability. The multi-user coding scheme design is a critical problem for UMA. This paper proposes a UMA coding scheme based on the T-Fold IRSA (irregular repetition slotted Aloha) paradigm by using joint Intra/inter-slot code design and optimization. Our scheme adopts interleave-division multiple access (IDMA) to enhance the intra-slot coding gain and the low-complexity joint intra/inter-slot SIC (successive interference cancellation) decoder structure to recover multi-user payloads. Based on the error event decomposition and density evolution analysis, we build a joint intra/inter-slot coding parameter optimization algorithm to minimize the SNR (signal-to-noise ratio) requirement at an expected system packet loss rate. Numerical results indicate that the proposed scheme achieves energy efficiency gain by balancing the intra/inter-slot coding gain while maintaining relatively low implementation complexity

A Specific and Sensitive Aptamer-Based Digital PCR Chip for <i>Salmonella typhimurium</i> Detection

Food poisoning and infectious diseases caused by Salmonella typhimurium (S. typhimurium) are serious public health concerns for human health and food safety. The diversity and complexity of food matrices pose great challenges for rapid and ultra-sensitive detection of S. typhimurium in food samples. A method capable of identification, detection, and quantification of S. typhimurium is essential for addressing these issues. In this study, aptamer-coated magnetic beads (Apt-MBs) are employed as capture bio-probes to specifically and selectively concentrate S. typhimurium in food samples. A self-priming chip-based digital PCR was then presented as another biosensor for on-site detection and quantification of S. typhimurium cells. The chip we developed was robust and did not require any external power for sample loading. The combination of Apt-MBs with an on-chip digital detection realized the integration into lab-on-a-chip-based biosensors for on-site monitoring of foodborne pathogens. It was possible to capture and detect S. typhimurium cells as low as 90 CFU/reaction with a capture efficiency of 94.5%. Additionally, the whole process only took about 2 h. This unique platform could also be used to monitor other target bacteria with high specificity and sensitivity by utilizing different aptamers. Furthermore, the platform has potential applications in point-of-care testing in the future

A Multiplex RT-PCR Assay for S. aureus, L. monocytogenes, and Salmonella spp. Detection in Raw Milk with Pre-enrichment

This study firstly developed a multiplex real-time PCR (RT-PCR) technique combined with a pre-enrichment step to simultaneously detect Staphylococcus aureus (S. aureus), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw milk and the dairy farm environment (feces, soil, feed, water) in one reaction. Brain heart infusion (BHI) broth was selected for the enrichment step to increase the density of the target bacteria by using an incubation of 4 h before multiplex RT-PCR. The results showed that the detection limit of the multiplex real-time assay was approximately 102 CFU/mL for pure cultures and artificially contaminated milk without enrichment, while 12, 14, and 10 CFU/25 mL, respectively, for S. aureus, L. monocytogenes, and Salmonella spp. after pre-enrichment. The newly developed multiplex RT-PCR assay was applied to 46 dairy farm environmental samples and raw milk samples covering a wide variety of sample types. The results demonstrated that the multiplex RT-PCR assay coupled with the BHI enrichment broth was suitable for the simultaneous screening of S. aureus, L. monocytogenes, and Salmonella spp. in the pasture environment and in raw milk. The multiplex RT-PCR assay clearly and successfully shortened the total detection time and reduced labor compared to conventional culture-based methods for testing natural samples

Multiple action sites of ultrasound on Escherichia coli and Staphylococcus aureus

Ultrasound, is thought to a potential non-thermal sterilization technology in food industry. However, the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure. In this study, the action modes of ultrasound on both Gram-negative and Gram-positive microorganisms were estimated. From colony results, ultrasound acted as an irreversible effect on both Eshcerichia coli and Staphylococcus aureus without sublethal injury. The result in this study also showed that a proportion of bacteria subpopulation suffered from serious damage of intracellular components (e.g. DNA and enzymes) but with intact cell envelopes. We speculated that the inactivated effects of ultrasound on microbes might more than simply completed disruption of cell exteriors. Those microbial cells who had not enter the valid area of ultrasonic cavitation might be injected with free radicals produced by ultrasound and experienced interior injury with intact exterior structure, and others who were in close proximity to the ultrasonic wave field would be immediately and completely disrupted into debris by high power mechanic forces. These findings here try to provide extension for the inactivation mechanisms of ultrasound on microorganisms. Keywords: Ultrasound, Inactivation mechanism, Reactive oxygen species (ROS), Staphylococcus aureus, Escherichia col