목차

1. Introduction 1
1.1 LAMP as an important molecular diagnostic tool 1
1.2 Retroref lect ive Janus particle as a non-spectroscopic optical sensing probe 2
1.3 Multiplex molecular diagnostic platform by integrating LAMP and retroreflective signaling 4
1.4 Aim of thesis 9
2. Materials and methods 11
2.1 Reagents and apparatus 11
2.2 Development of a new lamp method for forming double labeled amplicon 12
2.2.1 Introducing a new developed LAMP method 12
2.2.2 Primer design 16
2.2.3 Preparation of target genes for LAMP assay 18
2.2.4 LAMP assay 18
2.3 Fabrication of the sensing surface and optical probes 20
2.3.1 Modif icat ion of target-specif ic capture molecule on sensing substrate 20
2.3.2 Modification of Avidin on RJPs 23
2.4 Analysis of antibiotic-resistant Salmonella using the developed multiplex system 25
2.4.1 Quantitative analysis of antibiotic-resistant Salmonella 25
2.4.2 Quantitative analysis of specific amplicon from double-labeled amplicon mixture 27
3. Results and discussion 29
3.1 Optimization of the developed LAMP method 29
3.2 Confirmation of surface modification for target specific analysis 32
3.2.1 Verification of sensing surface 32
3.2.2 Verification of optical signaling probe 35
3.3 Optimization of double-labeled amplicon concentration and RJP reaction time for maximize the sensitivity 37
3.4 Quantitative analysis of antibiotic-resistant Salmonella 41
3.4.1 Calibration study of the developed platform for antibiotic-resistant Salmonella quantif ication 41
3.4.2 Calibration study of the developed platform using mixture of double labeled amplicon 46
4. Conclusions 49
5. References 51