Sample preparation has recently been an issue in the detection of food poisoning pathogens, particularly viruses such as norovirus (NoV), in food because of the complexity of foods and raw fresh materials. Here, we demonstrate a total analytical microfluidic chip module to automatically perform a series of essential processes (cell concentration, lysis (RNA extraction), nucleic acid amplification, and detection) for the fast but sensitive detection of norovirus in oysters. The murine NoV spiked oyster was stomached using a standard method. The supernatant was first loaded into a shape switchable sample preparation chamber consisting of charge switchable micro-beads. Murine NoV, which was adsorbed on microbeads by electrostatic physisorption, was lysed using bead beating. The extracted RNA was transferred to the detection chamber to be amplified using Nucleic Acid Sequence Based Amplification (NASBA). The optimal surface functionality, size, and number of microbeads were achieved for the virus concentration and the stable RNA extraction in the shape-switchable micro-channel. As a result, murine NoV in a single oyster was successfully detected within 4h by the microfluidic chip developed here, and could be directly applied to the large volume environmental sample as well as the food sample.
Keywords: Bead-beating; Charge switchable microbead; NASBA; Norovirus; Oyster; Shape switchable microchannel.
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