Methods and apparatus for point-of-care nucleic acid amplification and detection
First Claim
1. A method for point-of-care amplification and detection of a target nucleic acid sequence, the method comprising:
- collecting a sample matrix with filter paper;
advancing a punch element through the filter paper to form at least one punch of the filter paper;
transferring the at least one punch of the filter paper to a reaction chamber comprising nucleic acid amplification reagents;
sealing the at least one punch of the filter paper within the reaction chamber in fluid communication with the nucleic acid amplification reagents;
heating the nucleic acid amplification reagents with a heating element in order to amplify the target nucleic acid sequence when contained within the sample matrix;
detecting amplification of the target nucleic acid sequence with a sensor;
displaying detection results obtained with the sensor on a display; and
disposing of the at least one punch of the filter paper, the reaction chamber, the heating element, the sensor, the display and the nucleic acid amplification reagents after one-time use.
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Abstract
Methods and apparatus are provided for point-of-care nucleic acid amplification and detection. One embodiment of the invention comprises a fully integrated, sample-to-answer molecular diagnostic instrument that optionally may be used in a multiplexed fashion to detect multiple target nucleic acid sequences of interest and that optionally may be configured for disposal after one-time use. Optionally, sample preparation is fully or partially achieved via heat treatment and/or using a filter paper, such as a chemically treated filter paper, e.g., an FTA card. The instrument preferable utilizes an isothermal nucleic acid amplification technique, such as loop-mediated isothermal amplification (LAMP), to reduce the instrumentation requirements associated with nucleic acid amplification. Detection of target amplification may be achieved, for example, via detection of a color shift or fluorescence in a dye added to the amplification reaction.
40 Citations
15 Claims
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1. A method for point-of-care amplification and detection of a target nucleic acid sequence, the method comprising:
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collecting a sample matrix with filter paper; advancing a punch element through the filter paper to form at least one punch of the filter paper; transferring the at least one punch of the filter paper to a reaction chamber comprising nucleic acid amplification reagents; sealing the at least one punch of the filter paper within the reaction chamber in fluid communication with the nucleic acid amplification reagents; heating the nucleic acid amplification reagents with a heating element in order to amplify the target nucleic acid sequence when contained within the sample matrix; detecting amplification of the target nucleic acid sequence with a sensor; displaying detection results obtained with the sensor on a display; and disposing of the at least one punch of the filter paper, the reaction chamber, the heating element, the sensor, the display and the nucleic acid amplification reagents after one-time use. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification