Development of a rapid test kit for screening of infectious diseases

• Main Author: Tay, Oriana Ling Ying
• Other Authors: Bai Jianhao
• Format: Final Year Project
• Language: English
• Published: 2016
• Subjects: DRNTU::Science::Biological sciences
• Online Access: http://hdl.handle.net/10356/66962
• Institution: Nanyang Technological University

In point-of-care (POC) setting, lateral flow devices remain popular for the use in healthcare management. They are affordable, simple to use, disposable, capable of storage at room temperatures and enable visual interpretation of the test results. We have utilised a wash-step free ELISA lateral flow system into developing a POC device. This system improves the sensitivity of the device by use of an enzyme label, and reduces the sample volume required. However, the long shelf-life of an enzymatic substrate is typically achieved by storing as solution at 4°C (i.e. not at room temperature). The objective of this project is to determine the optimal conditions for storing the enzymatic substrate in dried form at room temperature for 2 years, and have a comparable assay sensitivity to the enzymatic substrate in solution form. To achieve this, we focus on 1) optimizing the enzymatic substrate concentration for drying and its long-term storage at room temperature, 2) determining the configuration of test strip layout, and 3) optimizing the resolubilization buffer solution used to rehydrate the dried enzymatic substrate. After optimization, testing of infectious diseases (Dengue NS1 and Hepatitis B surface antigen) in the POC device has shown to require only 5 µL of sample, rapid turnaround time of 15 minutes, and a limit-of-detection of 47.05 ng/mL and 20.52 ng/mL respectively.