Abstract
A nucleic acid amplification and detection device includes an amplification cartridge with a plurality of reaction chambers for containing an amplification reagent and a visual detection reagent, and a plurality of optically transparent view ports for viewing inside the reaction chambers. The cartridge also includes a sample receiving port which is adapted to receive a fluid sample and fluidically connected to distribute the fluid sample to the reaction chamber, and in one embodiment, a plunger is carried by the cartridge for occluding fluidic communication to the reaction chambers. The device also includes a heating apparatus having a heating element which is activated by controller to generate heat when a trigger event is detected. The heating apparatus includes a cartridge-mounting section which positioned a cartridge in thermal communication with the heating element so that visual changes to the contents of the reaction chambers are viewable through the view ports.
Claims
1. A nucleic acid amplification and detection device, comprising: an amplification cartridge having distribution chamber, a plurality of reaction chambers all arranged around and adjacent the distribution chamber in a hub-and-spoke configuration for containing an amplification reagent and a visual detection reagent and wherein the distribution chamber has a plurality of inlets leading to the respective reaction chambers, a plurality of optically transparent view ports for viewing inside the reaction chambers, a sample receiving port adapted to receive a fluid sample and fluidically connected to distribute the fluid sample to the reaction chambers via the distribution chamber, and a plunger positioned in the distribution chamber and actuable from the first position not occluding the inlets to a second position occluding the inlets for simultaneously occluding fluidic communication to all the reaction chambers; and a heating apparatus having a heating element, a controller adapted to activate the heating element to generate heat upon detecting a trigger even, and a cartridge-mounting section adapted to receive the amplification cartridge when loaded thereon so that the reaction chambers are in thermal communication with the heating element and so that visual changes to the contents of the reaction chambers are viewable through the view ports.
2. The nucleic acid amplification and detection device of claim 1, further comprising the amplification reagent and the visual detection reagent pre-loaded in the reaction chambers.
3. The nucleic acid amplification and detection device of claim 2, wherein the pre-loaded reagents are lyophilized.
4. The nucleic acid amplification and detection device of claim 1, wherein the cartridge-mounting section has a plural of heating wells in thermal communication with the heating element and adapted to receive therein the reaction chambers of the amplification cartridge.
5. The nucleic acid amplification and detection device of claim 4, wherein the heating element includes a plurality heating sub-elements each in thermal communication with a corresponding one of the heating wells.
6. The nucleic acid amplification and detection device of claim 1, wherein the heating apparatus includes a cover adapted to engage the cartridge-mounting section so as to cover a loaded amplification cartridge.
7. The nucleic acid amplification and detection device of claim 6, wherein the cover has view ports arranged to align with the view ports of the loaded amplification cartridge when the cover is engaged to cover the loaded amplification cartridge so that visual changes to the contents of the reaction chambers are viewable through the cover view ports.
8. The nucleic acid amplification and detection device of claim 6, wherein the trigger event is the engagement of the cover to cover the loaded amplification cartridge.
9. The nucleic acid amplification and detection device of claim 6, wherein the means for occluding fluidic communication to the reaction chambers is adapted to be activated by the cover upon engagement of the cover to cover the loaded amplification cartridge.
10. A multi-chamber, nucleic acid amplification cartridge for use with a heating apparatus of a type having a heating element and a cartridge-mounting section adapted to receive the amplification cartridge when loaded thereon, comprising; a cartridge body having a distribution chamber, a plurality of reaction chambers all arranged around and adjacent the distribution chamber in a hub-and-spoke configuration for containing an amplification reagent and a visual detection reagent and wherein the distribution chamber has a plurality of inlets leading to the respective reaction chambers, a plurality of optically transparent view ports for viewing inside the reaction chambers, and a sample receiving port adapted to receive a fluid sample and fluidically connected to distribute the fluid sample to the reaction chambers via the distribution chamber, wherein the reaction chambers are located on the cartridge body so as to be in thermal communication with the heating element when the amplification cartridge is loaded on the cartridge-mounting section of the heating apparatus, and wherein the view ports are located on the cartridge body so that the contents of the reaction chambers are viewable through the view ports when the amplification cartridge is loaded on the cartridge-mounting section of the heating apparatus; and a plunger positioned in the distribution chamber and actuable from a first position not occluding the inlets to a second position occluding the inlets for simultaneously occluding fludic communication to all the reaction chambers.
11. The multi-chamber nucleic acid amplification cartridge of claim 10, further comprising the amplification reagent and the visual detection reagent pre-loaded in the reaction chambers.
12. The multi-chamber nucleic acid amplification cartridge of claim 11, wherein the pre-loaded reagents are lyophilized.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1) The accompanying drawings, which are incorporated into and form a part of the disclosure, are as follows
(2) FIG. 1 is a perspective view of an example embodiment of the amplification cartridge of the present invention.
(3) FIG. 2 is a top schematic view of the amplification cartridge of FIG. 1 with the plunger assembly removed to view the distribution chamber.
(4) FIG. 3 is a radial cross-sectional view of the amplification cartridge of FIG. 2 taken along radial lines labeled “FIG. 3”, illustrating the flow of a fluid sample from the sample receiving port to one of the reaction chambers.
(5) FIG. 4 is a cross-sectional view of a plunger assembly of an example embodiment of the amplification cartridge of the present invention, shown in a first position not occluding/sealing the inlets to the reaction chamber.
(6) FIG. 5 is a cross-sectional view of the plunger assembly following FIG. 4, shown in a second position occluding/sealing the inlets to the reaction chamber.
(7) FIG. 6 is a perspective view of an example embodiment of the heating apparatus of the present invention.
(8) FIG. 7 is a perspective view of the amplification cartridge of FIG. 1 loaded in the heating apparatus of FIG. 6.
(9) FIG. 8 is a partial cross-sectional view of the amplification cartridge and heating apparatus of FIG. 7, showing a reaction chamber seated in a heating well of the heating apparatus.
(10) FIG. 9 is a perspective view of the amplification cartridge and heating apparatus of FIG. 8, shown with the cover engaged to the cartridge-mounting section to close the heating apparatus and cover the loaded amplification cartridge.
DETAILED DESCRIPTION
(11) Turning to the drawings, FIGS. 1-3 show an example embodiment of a multi-chamber, nucleic acid amplification cartridge of the present invention, generally indicated at reference character 10. As best shown in the perspective view of FIG. 1, the amplification cartridge has a cartridge body 11 (e.g. shown having a generally disc-shape), and a plunger assembly 17 located at the center of the cartridge body with a plunger 18 actuably carried on a plunger base mount 19 which may be connected to or an extension of the cartridge body. A plurality of reaction chambers 12 are also shown provided as cavities in the cartridge body, which are radially spaced from and arranged around the plunger assembly in a generally hub-and-spoke configuration, and which form cavity openings on one side (e.g. a top side) of the cavity body. And a sample receiving port 13 is also shown provided on the cartridge body for receiving a fluid sample (not shown) and distributing the fluid sample to each of the reaction chambers where it may mix with amplification and visual detection reagents which may be pre-loaded in the reaction chambers, or otherwise provided therein.
(12) The inside of the reaction chambers/cavities and its contents are viewable through optically transparent view ports/windows comprising optically transparent materials, such as glass, plastic, etc. which are arranged to sealably cover the reaction chamber/cavity openings. In FIGS. 1-3, the optically transparent materials are shown provided as a plurality of optically transparent window units 14, each sealably covering a cavity opening of a corresponding one of the reaction chambers 12. It is appreciated, in the alternative, that the optically transparent material may be provided as a single monolithic body (e.g. a ring-shaped structure) which sealably covers all reaction chamber openings, or two or more monolithic bodies, each sealably covering two or more reaction chamber openings.
(13) FIG. 2 also shows the amplification cartridge 10 of FIG. 1, but with the plunger assembly 17 removed to expose a distribution chamber 21 which is in fluidic communication with the sample receiving port 13 and each of the reaction chambers 12. In particular, a plurality of inlets 22 is shown in the distribution chamber 21 which allows fluid to pass out from the distribution chamber and into the reaction chambers via fluidic channels or conduits. FIG. 3 particularly shows the distribution path of a fluid sample introduced through the sample receiving port 13 to the reaction chambers. As shown by the arrows, a fluid sample is first introduced into the cartridge body 11 through sample receiving port 13 where it is transported radially inward to the distribution chamber 21 via fluidic conduit 20. From the distribution chamber, the fluid sample is passed into inlet 22, and transported radially outward via fluidic conduit 15, and into each of the reaction chambers, such as representative reaction chamber 12.
(14) Once the reaction chambers are loaded with the fluid sample, the plunger 18 of the plunger assembly may be actuated in the direction of the arrow in FIG. 3, i.e. toward the cartridge body, to occlude the inlets such as representative inlet 22. Inlet 22 is particularly shown with a seal 23 surrounding the inlet, which when brought into contact with the plunger 18 (i.e. when the plunger is actuated to a position which occludes fluidic communication between the distribution chamber 21 and the inlet 22 and reaction chamber 12) seals entry to or exit from the reaction chambers. The seal may be, for example, a raised collar, flange, O-ring, or other sealing material or structure. Backflow of a fluid sample may be prevented in this manner to isolate and prevent cross-contamination of the reaction chambers from each other. It is also appreciated that a separate diaphragm may be positioned adjacent the inlet 22 so as to be urged by the plunger against the inlets to occlude the inlets. When loaded into the reaction chambers, the fluid sample is viewable through the optically transparent view port 14, as well as a colorimetrically detectable reaction the fluid sample may have with amplification and detection agents contained in the chambers. FIGS. 1-3 also show expansion slots 16 fluidically connected to the reaction chambers for relieving pressure as fluid enters. Furthermore, an absorbent material, such as a sponge, may be provided for trapping excess fluid.
(15) FIGS. 4 and 5 show cross-sectional views of a plunger assembly of another example embodiment of the amplification cartridge of the present invention, shown in a first position in FIG. 4 not occluding/sealing the inlets to the reaction chamber, and in a second position in FIG. 5 occluding/sealing the inlets to the reaction chamber. The cartridge body 11 is shown having fluidic channels/conduits 15 terminating at inlets 22 in the distribution chamber 21. Seals 25 are also shown in the distribution chamber surrounding the inlets. And a plunger base mount 19 is shown having a lower cavity which together with the cavity body 11 forms the distribution chamber 21, and an upper cavity in which the center of the plunger is actuably positioned to be displaced into and out of the lower cavity. Also shown in the distribution chamber 21 is a sealing diaphragm 49 (e.g. made of a flexible or resiliently biasing material, such as rubber) held away from the inlets 22 by a spacer 28, which may be a large diameter O-ring, block, or other structure which suspends the sealing diaphragm in a non-occluding position as shown in FIG. 4. As shown between FIGS. 4 and 5, when the plunger 18 is actuated toward the cavity body 11, the plunger 18 urges the diaphragm 49 toward the inlets 2, until a sealing contact is made with the seals 25. The plunger base mount 19 is also shown having a guide track formed between walls 27 and 27′ in which a guide arm 26 of the plunger may be moved. In some embodiments, the guide arms 26 may be configured for a friction fit in the guide track such that the plunger 18 may remain in the occluding position once actuated.
(16) FIG. 6 shows an example embodiment of the heating apparatus of the present invention, generally indicated at 30, having a cartridge mounting section 31 that is adapted and configured to receive an amplification cartridge, such as 10 in FIG. 7, that may be loaded onto it for heating the reaction chambers, so that the reaction chambers of the cartridge are in thermal communication with a heating element (or sub-elements) and so that visual changes to the contents of the reaction chambers are viewable through the view ports. In FIG. 6, the mounting section 31 is shown having a centrally located raised platform 32 with a plurality of open cavities, i.e. heating wells 33, formed thereon, and with the heating element or sub-elements arranged to heat the wells. The raised platform is particularly shown having the same generally disk-shape of the amplification cartridge 10, with the heating wells 33 radially arranged to receive the reaction chambers into the cavities. For this purpose, and as shown in FIG. 3, the reaction chamber 12 may be formed in part by lower wall and floor 24 sections which protrude and otherwise extend below the cavity body 11, so that the protruding chambers may be seated in the heating wells 33 of the heating unit. This is best shown in FIG. 8 where the raised platform 32 and the wells 33 (i.e. cavity walls) are particularly shown integrally connected to perimeter walls 39 which surround and encase the raised platform 32 of the mounting section 31. FIG. 8 also shows one embodiment of the heating element 41 which is positioned on the outside of the heating wells and cavity walls 33 for heating the cavity and the reaction chamber 12 positioned therein. It is appreciated that the heating element may in the alternative be disposed in the cavities of the heating wells, or integrally formed in the walls of the heating wells, or provided as a single or multiple heating elements, with each heating element arranged to heat multiple reaction chambers together. It is also appreciated that the cartridge mounting section can be adapted to simply receive a cartridge, such as a form fitting space with no direct attachment or mounting mechanism, or in the alternative, be adapted to mount or releasably attach, hold, or secure a cartridge, such as by using a fastening mechanism such as for example clamps.
(17) FIGS. 6-9 also show included on the cartridge mounting section 31 a controller 34, including control electronics, such as for exampling a time circuit, for controlling reaction temperature and for controlling reaction start and stop times. An onboard power source, e.g. a battery not shown, may also be provided with the controller 34 for powering the controller, the heating element 41, and other electronics features provided on the heating unit. It is appreciated, however, that the heating unit may include connectors, such as a power cord and outlet plug, for connecting to an off-board power source, such as an electrical outlet. In this manner, the heating unit may be used to maintain an optimum reaction temperature(s) across the reaction chambers. Also shown provided is an activation switch 35 for initiating the controller to heat the reaction chambers. Optionally, an indicator light such as 36 may also be provided.
(18) As shown in FIGS. 6, 7, and 9 the heating unit 30 may also include a cover section or outer shell 37 for covering the mounting section 31 and any cartridge that may be positioned thereon. In particular, the cover section 37 is adapted to engage the cartridge-mounting section so as to cover a cartridge that is loaded on the cartridge mounting section, and is shown in particular hingedly connected to the mounting section 31 by hinge 40. The cover is shown having optically transparent windows 38 arranged to align with the optically transparent view ports 14 of a mounted cartridge 10, as shown in FIG. 9, for viewing reaction progress during heating. It is appreciated that the view ports on the cover may either be an opening or an optically transparent material. The cover 37 is also shown having a cutout 42 which allows the activation switch 35 and heating indicator light 36 to be viewable therethrough. For the switch, this enables manual activation of the heating operation after the cover is positioned to close the heating unit. In the alternative, the cutout 42 may be removed so that the cover contacts and activates the switch when closed over the mounting section 31, to automatically initiate heating.
(19) Although the description above contains many details and specifics, these should not be construed as limiting the scope of the invention but as merely providing illustrations of some of the presently preferred embodiments of this invention. Other implementations, enhancements and variations can be made based on what is described and illustrated in this patent document. The features of the embodiments described herein may be combined in all possible combinations of methods, apparatus, modules, systems, and computer program products. Certain features that are described in this patent document in the context of separate embodiments can also be implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single embodiment can also be implemented in multiple embodiments separately or in any suitable subcombination. Moreover, although features may be described above as acting in certain combinations and even initially claimed as such, one or more features from a claimed combination can in some cases be excised from the combination, and the claimed combination may be directed to a subcombination or variation of a subcombination. Similarly, while operations are depicted in the drawings in a particular order, this should not be understood as requiring that such operations be performed in the particular order shown or in sequential order, or that all illustrated operations be performed, to achieve desirable results. Moreover, the separation of various system components in the embodiments described above should not be understood as requiring such separation in all embodiments.
(20) Therefore, it will be appreciated that the scope of the present invention fully encompasses other embodiments which may become obvious to those skilled in the art. In the claims, reference to an element in the singular is not intended to mean “one and only one” unless explicitly so stated, but rather “one or more.” All structural and functional equivalents to the elements of the above-described preferred embodiment that are known to those of ordinary skill in the art are expressly incorporated herein by reference and are intended to be encompassed by the present claims. Moreover, it is not necessary for a device to address each and every problem sought to be solved by the present invention, for it to be encompassed by the present claims. Furthermore, no element or component in the present disclosure is intended to be dedicated to the public regardless of whether the element or component is explicitly recited in the claims. No claim element herein is to be construed under the provisions of 35 U.S.C. 112, sixth paragraph, unless the element is expressly recited using the phrase “means for.”
