Ultrafast Pcr Microfluidic Chip Combined with Paper-Based Immunochromatographic and Triple-Stranded Dna (Tfo) Detection Technique for Rapid Test of Bacterial and Virus

The goal of this proposal is to develop an ultra-fast (＜ 3 min) PCR microfluidic chip and to integrate two detection technologies for different application scenario. For example, for the point-of care application, immunochromatographic test will integrate with ultra-fast PCR to provide a fast and eye-reading molecular diagnosis. In addition, for the high sensitivity detection, triplex-forming oligonucleotide label technique will integrate with ultra-fast PCR to provide a fast and high sensitivity molecular diagnosis. PCR is wildly used in molecular diagnosis, in which the global market is over 5 billion in USD. However, currently used PCR spends too much time, including some microfluidic type rapid PCR (several tens minutes). The most important keys for ultra-fast PCR are the proper reagent and rapid temperature control system. It is known that the high enzyme and oligonucleotide concentration is suitable for the ultra-fast PCR. However, there is no commercially available instrument or microfluidic chip which can provide such rapid temperature control. Microfluidic technology has the advantages of tiny sample volume (small thermal capacity) and high surface to volume ratio (high thermal conductivity). It is very suitable for the development of ultra-fast PCR system. In the first year, an ultra-fast PCR microfluidic chip which can perform 30-40 cycles within 3 min will develop. The observation of the PCR product mostly relies on gel electrophoresis, which is a time consumption and bulky instrument setup. In the second year, the ultra-fast PCR will integrate with immunochromatographic test to provide a rapid molecular diagnosis microfluidic system, which can be done within 10 min. In the third year, the ultra-fast PCR will integrate with a novel DNA label technique, triplex-forming oligonucleotide, which can provide a rapid and high sensitivity detection. We believe that with these new technologies the molecular diagnosis can be applied to point of care scenario.

Project ID:PB10901-3460
External Project ID:MOST108-2221-E182-012-MY3