Proceedings of the
9th International Conference of Asian Society for Precision Engineering and Nanotechnology (ASPEN2022)
15 – 18 November 2022, Singapore
doi:10.3850/978-981-18-6021-8_OR-03-0082
Development of A Microfluidic Chip for Early And Rapid Detection of Multiple Dengue Serotypes
1Smart Microfluidics (SMF), Singapore Institute of Manufacturing Technology, Nanos, 31 Biopolis Way, #04-10, S(138669), Singapore
2Printed Intelligent Device (PID), Singapore Institute of Manufacturing Technology, 73 Nanyang Dr, S(637662), Singapore
ABSTRACT
It is important to distinguish between the 4 dengue virus serotypes, DEN 1, DEN 2, DEN 3 and DEN 4, when one is infected with a dengue fever episode due to lack of cross-protective neutralizing antibodies for each serotype. Secondary infections also put individuals at higher risks for severe dengue illness than those who have not been previously infected. Current gold-standard assays include reverse transcription-PCR (RT-PCR) and other laboratory and specialized labor-intensive assays like nucleotide sequencing and serotype-specific ELISA. To enable on-field diagnosis of dengue serotypes, the detection process would need to be simplified and made at least semi-automated. Polymer-based microfluidics manufacturing methods had been employed for the fabrication of such point-of-care diagnostic devices, particularly on sample loading, cell lysis, RNA extraction and RT-PCR. A downstream detection module was conceptualized and fabricated to detect the amplified DNA from provided PCR mix (product) in that sample. Further, to ensure accuracy, each serotype assay necessitates a positive control and a negative control, which constitutes 3 separate channels for diagnosis of just 1 serotype. In this study, a 6-channel bi-assay microfluidic chip was designed with pre-loaded diluent (ultrapure water) and cyanine dye, sample chamber for loading, sequential fluidic sample mixing, and integrated membranes for simultaneous (6-channel) fluidic manipulation from a single actuation source. Positive samples turned the dye from blue to violet while the negative controls remained blue. The integrated membranes provided color contrast and facilitated the manipulation of the samples to the same line of sight for simultaneous analysis, paving the way for automated color analysis via smartphone.
Keywords: Microfluidics, Chip Design, Dengue Serotypes.
1Smart Microfluidics (SMF), Singapore Institute of Manufacturing Technology, Nanos, 31 Biopolis Way, #04-10, S(138669), Singapore
2Printed Intelligent Device (PID), Singapore Institute of Manufacturing Technology, 73 Nanyang Dr, S(637662), Singapore
ABSTRACT
It is important to distinguish between the 4 dengue virus serotypes, DEN 1, DEN 2, DEN 3 and DEN 4, when one is infected with a dengue fever episode due to lack of cross-protective neutralizing antibodies for each serotype. Secondary infections also put individuals at higher risks for severe dengue illness than those who have not been previously infected. Current gold-standard assays include reverse transcription-PCR (RT-PCR) and other laboratory and specialized labor-intensive assays like nucleotide sequencing and serotype-specific ELISA. To enable on-field diagnosis of dengue serotypes, the detection process would need to be simplified and made at least semi-automated. Polymer-based microfluidics manufacturing methods had been employed for the fabrication of such point-of-care diagnostic devices, particularly on sample loading, cell lysis, RNA extraction and RT-PCR. A downstream detection module was conceptualized and fabricated to detect the amplified DNA from provided PCR mix (product) in that sample. Further, to ensure accuracy, each serotype assay necessitates a positive control and a negative control, which constitutes 3 separate channels for diagnosis of just 1 serotype. In this study, a 6-channel bi-assay microfluidic chip was designed with pre-loaded diluent (ultrapure water) and cyanine dye, sample chamber for loading, sequential fluidic sample mixing, and integrated membranes for simultaneous (6-channel) fluidic manipulation from a single actuation source. Positive samples turned the dye from blue to violet while the negative controls remained blue. The integrated membranes provided color contrast and facilitated the manipulation of the samples to the same line of sight for simultaneous analysis, paving the way for automated color analysis via smartphone.
Keywords: Microfluidics, Chip Design, Dengue Serotypes.
