Microfluidic device for in-field purification, amplification, and detection of dna from mycotoxin-producing fungi
AssigneePurdue Research Foundation
InventorsJoseph Paul Robinson, Bartlomiej P. Rajwa, Euiwon Bae, Cole Reynolds, Xi Wu
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Abstract
Methods of testing grain samples for the presence of mycotoxin-producing fungi using portable devices are disclosed. The design and fabrication of such devices to practice the disclosed methods allow for rapid and portable in-field detection of mycotoxin-producing fungi prior to harvest and potential contamination of the food supply with infected grain.
Claims19
A method of detecting the presence of mycotoxin-producing fungus in grain in the field, comprising:
- a) procuring a grain sample;
- b) extracting DNA from the grain sample;
- c) isothermally amplifying the extracted DNA;
- d) identifying any DNA from mycotoxin producing fungus as amplified target DNA;
- and
- e) fluorescing products of amplified target DNA reactions to yield quantitative information relating to the presence of DNA from mycotoxin-producing fungus;
- wherein steps c-e are all performed within the same microfluidic chamber platform.
A method of in-field detection of mycotoxin producing fungus, comprising:
- a) collecting potentially infected plant tissue samples;
- b) grinding potentially infected plant tissues to yield ground tissue material;
- c) lysing ground tissue material to extract DNA;
- d) purifying extracted DNA to yield purified DNA;
- e) performing a fluorescence-based loop-mediated isothermal amplification assay on the purified DNA;
- f) measuring the amount of fluorescence from DNA from mycotoxin-producing fungus to yield a quantitative measurement;
- wherein steps b-c are performed in a first device and steps d-f are performed in a second device.
A portable microfluidic device for extracting fungal DNA from grain samples and performing qualitative and quantitative analysis of the same, comprising:
- a housing;
- a bead-bashing chamber enclosed within the housing for extracting DNA from plant tissue samples;
- a microfluidic chamber for purifying extracted DNA and performing a fluorescence-based loop-mediated isothermal amplification assay on the purified DNA positioned within the housing and operationally connected to the bead-bashing chamber;
- and
- a power source positioned within the housing and operationally connected to the microfluidic chamber.
Dependencies are inferred from the claim text.