Development of a non-spectroscopic biosensing kit for the quantitative analysis of foodborne pathogenic bacteria
- 주제(키워드) Foodborne pathogen , Salmonella Typhimurium , Retroreflection , Immunosensing
- 주제(DDC) 547
- 발행기관 아주대학교
- 지도교수 윤현철
- 발행년도 2023
- 학위수여년월 2023. 8
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000032855
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
As the food industry expands and the number of consumers increases, food safety is becoming more important worldwide. However, the outbreak of food poisoning caused by foodborne pathogenic bacteria is a great threat to food safety. Salmonella is one of the major causes of foodborne illness. However, conventional methods such as the selective culture method, polymerase chain reaction (PCR), and enzyme-linked immunosorbent assay (ELISA) for detecting foodborne pathogenic bacteria are time-consuming, labor-intensive, and require complicated equipment to measure. Thus, we developed a microfluidic chip for detecting pathogens rapidly, accurately, and sensitively. The form of the chip was created based on the lateral flow immunoassay (LFIA) commonly used for rapid diagnosis. The sensing surface is composed of two capture areas, a test area that captures immune complex of pathogens and particles and a control area that captures the residual particles not reacted with target pathogens. By applying the form of LFIA to a microfluidic chip and using retro-reflective particles as a signal probe, it was possible to simultaneously perform a high-sensitivity quantitative analysis of foodborne pathogens and judge the effectiveness of the chip. Furthermore, by using the characteristics of retro-reflective particles even in non-spectral light, it was possible to detect bacteria with simple optical equipment. In this study, the concentration of Salmonella Typhimurium was measured ranging from 0 to 103 CFU/mL. As a result, we could rapidly detect Salmonella Typhimurium up to the limit of detection to 10 CFU/mL.
more목차
1. Introduction 1
1.1 The risk of food poisoning by pathogenic bacteria 1
1.2 Conventional methods for pathogenic bacteria detection 2
1.3 Sensing strategy of the developed bacteria detection system 3
1.4 Aims of thesis 11
2. Materials and methods 12
2.1 Reagents and apparatus 12
2.2 Fabrication of sensing surface and optical signaling probe 13
2.2.1 Functionalization of the glass surface 13
2.2.2 Utilization of RJP as an optical sensing probe 16
2.3 Preparation of Salmonella bacteria sample 18
2.4 Designing of the sensing surface for quantitative immunoassay 21
3. Results and discussion 23
3.1 Verification of sensing surface and optical signaling probe 23
3.2 Immunoaffinity test of anti-Salmonella polyclonal antibody to target pathogen 27
3.3 Feasibility test of antigen-antibody based sandwich type Salmonella immunoassay 30
3.4 Verification of the new developed chip sensing surface 34
3.5 Quantitative analysis according to Salmonella concentration 37
3.6 Image processing for quantitative analysis 42
4. Conclusions 44
5. References 45
