Article Sharing| MIRA Technology for Detection of Norovirus Using a Visualization Device

Experimental Method
Cellulose paper strips absorbed MNV-1 RNA, which was then released into the MIRA reagent. The RNA was first reverse-transcribed into complementary DNA (cDNA) and then amplified into double-stranded DNA (dsDNA). Subsequently, probes specifically bound to the target sequence. After the tetrahydrofuran (THF) residue was cleaved by endonuclease IV, the remaining strand was extended by polymerase to produce a complete amplicon. The amplicon was captured and visualized as a color change on the T-line.

(Schematic Diagram of Experimental Principle)

Development Achievements

(Schematic Diagram of Paper/PDMS Microfluidic Device)

The research team developed a foldable paper/PDMS microfluidic device that integrates multiple functions, including nucleic acid extraction, isothermal nucleic acid amplification, and visual detection. This device seamlessly integrates the entire process, from sample preparation to the endpoint detection of MNV-1.

The microfluidic device completes the detection within 35 minutes, with a detection limit of 200 PFU/mL. The cost of each microfluidic device is calculated to be $6.76, making it the most cost-effective method among all known detection methods used by the research team.

Experimental Significance

The research team further evaluated the performance of this microfluidic device for detecting MNV-1 in lettuce and raspberries. They introduced crude MNV-1 RNA extracts into fresh agricultural samples, eliminating the need for complex virus RNA extraction and purification steps. The device detected 170 PFU/g in lettuce and 230 PFU/g in raspberries.

(Sensitivity of MNV-1 Detection in Lettuce and Raspberries Using the Microfluidic Device)

Traditional molecular methods for detecting viruses in food, such as PCR and qPCR, require high-purity viruses, which makes the process of virus elution, concentration, and purification from food matrices complex. Additionally, traditional methods require complicated, bulky, and expensive equipment, making them unsuitable for use in resource-limited environments.

This device holds significant value for the rapid detection of HuNoV and other foodborne viruses within the food supply chain.

Article Information
Journal: Applied and Environmental Microbiology
Article Title: Hybrid paper/PDMS microfluidic device integrated with RNA extraction and recombinase polymerase amplification for detection of norovirus in foods
Impact Factor: 3.9
Research Team: Viral Diseases Center, Lady Davis Institute, McGill University