Performing Department
(N/A)
Non Technical Summary
Getting sick from eating contaminated food is a risk we all face. Illnesses caused by bacteria in our food are becoming a big health and economic problem in the United States. As these bacteria become resistant to antibiotics, detecting and treating the illnesses they cause are getting even harder. Currently, the best ways to detect these antibiotic-resistant bacteria in food are through slow methods like culturing and PCR (polymerase chain reaction). While accurate, these methods take a long time and require trained experts to operate. This project aims to create a small, portable device called the Nanophotonic-CRISPR Chip (NPC-Chip) that can quickly and accurately detect antibiotic-resistant bacteria in food. No expert or lab is needed. The NPC-Chip combines light and CRISPR technologies into one tiny chip. Non-expert users can easily carry and use it to identify the contamination in food. In this project, the chip performance will be tested in milk samples. If successful, this fast, sensitive, and portable NPC-Chip can help control the spread of disease-causing bacteria in our food supply chain and reduce the overuse of antibiotics, crucial for food safety and public health.?
Animal Health Component
50%
Research Effort Categories
Basic
40%
Applied
50%
Developmental
10%
Goals / Objectives
The major goal of this project is to create a small and portable device called the Nanophotonic-CRISPR Chip (NPC-Chip) that can quickly and reliably detect antibiotic-resistant bacteria in food without needing experts or a lab. Towards that, the project encompasses two specific objectives to implement in two years as summarized below:Objective 1. Design, Fabrication, and Integration of Nanophotonic-CRISPR Chip (NPC-Chip)Objective 2. Assessment and Optimization of NPC-Chip to detect foodborne bacterial pathogens
Project Methods
The project will employ a multidisciplinary approach combining nanophotonic design, CRISPR-based assays, and biosensor development to create the Nanophotonic-CRISPR Chip (NPC-Chip) for rapid and sensitive detection of foodborne pathogens, including antibiotic-resistant strains.1. Nanophotonic Design and Fabrication:o Simulation and modeling will be used to design the nanophotonic sensing structure based on the principle of Electromagnetic Induced Transparency (EIT), which provides narrow spectral linewidths and enhanced sensitivity.o Standard microfabrication techniques, such as photolithography and dry etching, will be employed to fabricate the designed nanophotonic structures on silicon chips.2. CRISPR-based Assay Development:o Isothermal recombinase polymerase amplification (RPA) will be used for amplifying bacterial DNA targets.o CRISPR-Cas effectors will be designed and optimized for specific and accurate recognition of antibiotic-resistant bacterial DNA sequences.o Solid-phase CRISPR assays will be developed, where the CRISPR complex cleaves gold nanoparticle-labeled reporter probes immobilized on the nanophotonic chip upon target recognition.3. Integration and Optimization:o The nanophotonic sensing structures and CRISPR-based assays will be integrated to create the NPC-Chip.o Microfluidic components will be incorporated to enable the delivery of reagents and samples to the chip.o Extensive testing and optimization will be conducted to evaluate the sensitivity, specificity, and performance of the NPC-Chip in detecting various foodborne pathogens and their antibiotic-resistant variants.4. Evaluation and Data Analysis:o The sensitivity of the NPC-Chip will be evaluated using serial dilutions of purified bacterial DNA samples, with results validated by standard PCR and culturing assays.o The specificity and cross-reactivity of the NPC-Chip will be assessed by testing against various bacterial strains, including antibiotic-resistant and wild-type strains.o The performance of the NPC-Chip will be evaluated using spiked milk samples to simulate real-world food matrices.o Statistical analyses will be performed to quantify the sensitivity, specificity, and accuracy of the NPC-Chip in detecting foodborne pathogens.Efforts:1. Formal Classroom Instruction:o The PD and co-PD will incorporate the knowledge developed in this project into their respective courses.o Develop course modules related to nanotechnology, biosensors, food safety, and pathogen detection.2. Laboratory Instruction:o These laboratory instructional sessions will provide opportunities for experiential learning and skill development in areas such as nanophotonic design, CRISPR-based assays, and biosensor development.o Hands-on training in laboratory settings will be integrated with current curricula to teach students and researchers the sensing principles and practical aspects involving NPC-Chip technology.3. Extension and Outreach:o The project team will engage with local community organizations and high schools to provide hands-on training in laboratory settings.o This outreach effort can be collaborated with the ASU SCENE program, where the PI is an active mentor, to offer experiential learning opportunities for students and community members.o These activities will not only disseminate knowledge about the NPC-Chip technology but also inspire and encourage interest in STEM fields among diverse audiences.4. Public Workshops:o Public workshops and NPC-Chip demonstration sessions will be offered to expose the general public to the technology and its potential applications.o These workshops can be held at events such as the Arizona Science Fair and ASU Open Day, which attract visitors from across the nation.o These public workshops will aim to raise awareness, generate interest, and foster a broader understanding of the NPC-Chip technology and its significance in food safety and public health.Evaluation:1. Sensitivity and Specificity Evaluation:o Quantify the detection limit of the NPC-Chip for various foodborne pathogens and their antibiotic-resistant variants using serial dilutions of purified bacterial DNA samples.o Evaluate the specificity of the NPC-Chip by testing its ability to distinguish between antibiotic-resistant strains, wild-type strains, and other bacterial species.o Compare the sensitivity and specificity of the NPC-Chip with established methods, such as PCR and culturing assays.2. Performance Evaluation in Food Matrices:o Assess the performance of the NPC-Chip in detecting foodborne pathogens in spiked milk samples, simulating real-world food matrices.o Quantify the accuracy, reproducibility, and robustness of the NPC-Chip in detecting pathogens in complex food samples.o Evaluate the potential scalability and large-volume sampling for the use of NPC-Chip.3. Milestone Tracking:o Monitor and evaluate the progress of the project against predefined milestones and deliverables, such as successful nanophotonic design, CRISPR assay development, and NPC-Chip integration, through regular team meetings, progress reports, and reviews.o Track the achievement of key milestones, such as successful fabrication and testing of the nanophotonic sensing structures, optimization of the CRISPR-based assays for target detection, and integration of the nanophotonic and CRISPR components into the final NPC-Chip prototype.o Assess the project's adherence to the proposed timeline and identify any potential delays or challenges that may require adjustments or contingency plans.4. Impact Assessment:o Evaluate the effectiveness of the NPC-Chip technology by comparing its performance with existing methods in market.o Based on above evaluation, assess the potential economic and public health benefits of the NPC-Chip technology.o Assess the impact and collect feedback of the NPC-Chip technology on improving food safety practices through our efforts such as outreach activities and experiential learning opportunities and public workshops.