Methods and apparatus for point-of-care nucleic acid amplification and detection

摘要

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. 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. Such detection may be performed visually by an operator or may be achieved utilizing an imaging technique, e.g., spectrophotometric imaging.

主权项

1. A method for amplification and detection of a target nucleic acid sequence in a sample, the method comprising: transferring the sample through an inlet into a diagnostic apparatus; distributing the sample among a plurality of microfluidic channels of the diagnostic apparatus, each microfluidic channel extending to one of a plurality of reaction chambers in the diagnostic apparatus, at least one of the reaction chambers having a nucleic acid amplification reagent therein, so that a portion of the sample is received in each reaction chamber; heating the reaction chambers in order to amplify any target nucleic acid sequence within the sample portion; manually sliding a locking valve within a void of the diagnostic apparatus from an open position to a closed position after the transferring step to block all of the microfluidic channels and prevent backflow of the sample portion from each reaction chamber through the microfluidic channel extending to it during heating; and detecting in the at least one reaction chamber amplification of the target nucleic acid sequence within the sample.