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

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. 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.

What is claimed is: 
     
         1 . Apparatus for point-of-care amplification and detection of a target nucleic acid sequence, the apparatus comprising:
 a sample collector configured to collect a sample matrix;   a device having at least one microfluidic channel, at least one reaction chamber with nucleic acid amplification reagents, and at least one valve; and   a heating element,   wherein the at least one microfluidic channel is configured to transport the sample matrix from the sample collector to the at least one reaction chamber, wherein the heating element is configured to heat the nucleic acid amplification reagents after transport of the sample matrix to the at least one reaction chamber in order to amplify the target nucleic acid sequence when contained in the sample matrix, and wherein the at least one valve is configured to prevent backflow of the sample matrix from the at least one reaction chamber through the at least one microfluidic channel during heating.   
     
     
         2 . The apparatus of  claim 1 , wherein the device further comprises at least one sequestered chamber for venting of overflow from the at least one reaction chamber. 
     
     
         3 . The apparatus of  claim 2 , wherein the device further comprises flow control media for venting of gases but not liquids from the at least one reaction chamber. 
     
     
         4 . The apparatus of  claim 1 , wherein the valve is chosen from the group consisting of one-way valves, locking valves, reversible valves, irreversible valves and combinations thereof.