Nucleic Acid Amplification and Detection Devices, Systems and Methods

Abstract

The instant disclosure provides nucleic acid amplification systems and multi-reaction analysis systems useful in the efficient processing of samples, including clinical samples. Integrated systems that include nucleic acid amplification devices functionally combined with multi-reaction analysis systems are also included. Also provided are methods for monitoring multiple concurrent nucleic acid amplification reactions that include the use of devices and systems described herein.

1 . A thermal block for simultaneous nucleic acid amplification and reaction analysis; the thermal block comprising:
 a) at least two reaction vessel wells, each well comprising:
 i) a top opening configured to receive a reaction vessel inserted vertically into the well; 
 ii) a side aperture configured to allow light to pass laterally into the reaction vessel well, wherein upon insertion of a reaction vessel into the well a majority of the sidewall of the reaction vessel is in thermal contact with the well and a portion of the sidewall is exposed to light by the aperture; 
   b) a thermal transfer surface opposite the side apertures of the two reaction vessel wells; and   c) a mounting hole positioned between the at least two reaction vessel wells and having a center axis perpendicular to the plane of the thermal transfer surface.   
     
     
         2 . The thermal block of  claim 1 , wherein the block is bilaterally symmetrical along a vertical axis. 
     
     
         3 . The thermal block of  claim 1 , wherein the thermal transfer surface is essentially flat. 
     
     
         4 . The thermal block of  claim 1 , wherein the mounting hole is positioned equidistant from the two reaction vessels. 
     
     
         5 . The thermal block of  claim 4 , wherein the mounting hole is centrally positioned between the top and bottom sides of the thermal block. 
     
     
         6 . The thermal block of  claim 4 , wherein the mounting hole is centrally positioned between the right and left sides of the thermal block. 
     
     
         7 . The thermal block of  claim 1 , wherein the top surface of the thermal block comprises a raised flange encircling the circumference of each of the two reaction vessel wells. 
     
     
         8 . The thermal block of  claim 1 , wherein the surface of the thermal block opposite the thermal surface comprises a plurality of raised ridges. 
     
     
         9 . The thermal block of  claim 8 , wherein the plurality of raised ridges emanate radially from the mounting hole. 
     
     
         10 . The thermal block of  claim 1 , wherein the thermal block is constructed of aluminum. 
     
     
         11 . The thermal block of  claim 1 , wherein at least the two reaction vessel wells are nickel plated. 
     
     
         12 . The thermal block of  claim 11 , wherein at least a portion of the thermal block other than the two reaction vessel wells is nickel plated. 
     
     
         13 . The thermal block of  claim 12 , wherein the entire thermal block is nickel plated. 
     
     
         14 . The thermal block of  claim 1 , wherein the two reaction vessel wells comprise a lubrication coating. 
     
     
         15 . The thermal block of  claim 14 , wherein the lubrication coating is a dry lubrication coating. 
     
     
         16 . The thermal block of  claim 1 , wherein the thermal block further comprises a temperature detection area configured for the functional attachment of a temperature detector positioned proximally to each reaction vessel well. 
     
     
         17 . The thermal block of  claim 16 , wherein the temperature detection areas are positioned below the reaction vessel wells. 
     
     
         18 . The thermal block of  claim 1 , wherein each reaction vessel well further comprises a basal reservoir configured such that upon insertion of the reaction vessel into the well the reaction vessel does not contact the bottom of the well. 
     
     
         19 . The thermal block of  claim 1 , wherein the thermal block has a mass of 2 to 4 grams. 
     
     
         20 . A nucleic acid amplification module, the module comprising:
 a) a thermoelectric cooler unit comprising a mounting hole;   b) a thermal block of  claim 1 , wherein the thermal transfer surface is in thermal contact with a first surface of the thermoelectric cooler unit; and   c) a heatsink configured to receive a mechanical fastener, wherein the heatsink is in thermal contact with a second surface of the thermoelectric cooler unit and the mounting holes are aligned such that the thermal block, thermoelectric cooler unit, and the heatsink are joined by a mechanical fastener positioned through the mounting holes and affixed to the heatsink.   
     
     
         21 . The module of  claim 20 , further comprising a conductive pad between the thermal block and the thermoelectric cooler unit or between the thermoelectric cooler unit and the heatsink. 
     
     
         22 . The module of  claim 21 , wherein the conductive pad is a graphite pad. 
     
     
         23 . The module of  claim 20 , wherein the module comprises conductive pads both between the thermal block and the thermoelectric cooler unit and between the thermoelectric cooler unit and the heatsink. 
     
     
         24 . The module of  claim 20 , wherein the mechanical fastener joins the thermal block, thermoelectric cooler unit, and the heatsink by a compression force. 
     
     
         25 . The module of  claim 24 , wherein the mechanical fastener is a compression screw. 
     
     
         26 . The module of  claim 24 , wherein the compression force is between 100 and 200 pounds per square inch (psi). 
     
     
         27 . The module of  claim 20 , wherein the thermal block and the thermoelectric cooler are supported by a support bar fastened to the heatsink. 
     
     
         28 . The module of  claim 20 , wherein the module further comprises a heatsink fan configured to force air past the heatsink. 
     
     
         29 . The module of  claim 28 , wherein the heatsink is joined to the heatsink fan by a duct. 
     
     
         30 . The module of  claim 20 , wherein the thermal block has an operating thermal slew rate of greater than 5° C. per second. 
     
     
         31 . The module of  claim 20 , wherein the module further comprises one or more resistance thermometers (RTDs) in thermal contact with the thermal block. 
     
     
         32 . The module of  claim 31 , wherein the module comprises two RTDs in thermal contact with the thermal block, wherein each of the two RTDs are in proximity with a reaction vessel well of the thermal block. 
     
     
         33 . The module of  claim 31 , wherein the thermal contact between the one or more RTDs and the thermal block is maintained by a cantilever bar comprising one or more cantilever arms. 
     
     
         34 . The module of  claim 20 , wherein the module comprises an attached printed circuit board (PCB) for monitoring or controlling at least one electrical component of the module. 
     
     
         35 . The module of  claim 34 , wherein the PCB is conformal coated. 
     
     
         36 . The module of  claim 31 , wherein the module comprises at least one RTD in thermal contact with the thermal block and electrically connected to the PCB. 
     
     
         37 . The module of  claim 20 , wherein the module comprises a RTD in thermal contact with the thermal block or the heatsink, wherein the RTD in thermal contact with the thermal block or the heatsink is configured to monitor the temperature of the thermal block or heatsink and trigger a cutoff of power to the thermal block or heatsink if the temperature indicates a thermal error condition. 
     
     
         38 . The module of  claim 20 , wherein the heatsink is configured to receive a second mechanical fastener and the nucleic acid amplification module further comprises a second thermal block in thermal contact with a second thermoelectric cooler unit in thermal contact with the heatsink, wherein the second thermal block, the second thermoelectric cooler unit and the heatsink are joined by a second mechanical fastener positioned through mounting holes in the second thermal block and the second thermoelectric cooler unit and affixed to the heatsink. 
     
     
         39 . The module of  claim 38 , wherein the first thermal block and the second thermal block comprise separate electrical connections and are controlled independently. 
     
     
         40 . The module of  claim 20 , wherein the module further comprises one or more reaction vessel clamping bars. 
     
     
         41 . The module of  claim 40 , wherein the one or more reaction vessel clamping bars provides a compression force on reaction vessels within the reaction vessel wells of 5 N or more. 
     
     
         42 - 72 . (canceled)