CN116669854A

CN116669854A - System and method for advancing reactions between multiple chambers of a test device

Info

Publication number: CN116669854A
Application number: CN202180075306.1A
Authority: CN
Inventors: 托马斯·E·肖斯; 尹鹏
Original assignee: Harvard University Board Of Trustees
Current assignee: Harvard University Board Of Trustees
Priority date: 2020-11-12
Filing date: 2021-11-12
Publication date: 2023-08-29

Abstract

A test apparatus includes an elongate member and a tube assembly. The tube assembly has a first end and a second end. The tube assembly is configured to receive the elongate member at the second end. The tube assembly includes a plurality of chambers including a first chamber and a second chamber. The first chamber and the second chamber are separated by a membrane. The tube assembly also includes a spring at the second end of the tube assembly. The tube assembly also includes a spring retainer configured to prevent decompression of the spring when in the locked position and to allow decompression of the spring when in the unlocked position.

Description

System and method for advancing reactions between multiple chambers of a test device

Cross Reference to Related Applications

The present application claims priority and benefit from the following patent applications, and the following patent applications are incorporated herein by reference: U.S. provisional patent application No. 63/112,751, filed 11/12 in 2020; U.S. provisional patent application No. 63/124,919 filed on 12/14 2020; U.S. provisional patent application No. 63/126,701 filed on 12/17 2020; U.S. provisional patent application No. 63/191,205 filed on day 20, 5, 2021; and U.S. provisional patent application No. 63/270,350 filed on day 21, 10, 2021.

Statement regarding federally sponsored research

The present invention was carried out with government support under GM133052 awarded by the national institutes of health. The government has certain rights in the invention.

Technical Field

The present invention relates generally to apparatus and methods for testing samples. In particular, the invention relates to a testing device having a plurality of chambers, the testing device advancing at least part of an elongate member between a first chamber and a second chamber of the testing device.

Background

Pandemics or epidemics requiring a large number of tests (e.g., a covd-19 pandemic) place strains on test resources based on a centralized test infrastructure, such as involving a small number of test laboratories or centers when providing large numbers of test results to a population. By eliminating any bottlenecks associated with the centralized infrastructure, the popularity or dispersion of testing can greatly improve efficiency. For example, test results may be provided to a population more quickly in a more decentralized test environment. However, such decentralized testing requires publicly available materials and equipment, which may not be familiar with the testing process. Accordingly, the present disclosure relates to error proofing test equipment for various applications.

Disclosure of Invention

According to some embodiments of the present invention, an apparatus for testing is provided. The apparatus includes an elongate member and a tube assembly. The tube assembly has a first end and a second end. The tube assembly is configured to receive the elongate member at the second end. The tube assembly includes a plurality of chambers including a first chamber and a second chamber. The first chamber and the second chamber are separated by a membrane. The tube assembly also includes a spring at the second end of the tube assembly. The tube assembly also includes a spring retainer configured to prevent decompression of the spring when in the locked position and to allow decompression of the spring when in the unlocked position.

According to some embodiments of the present disclosure, a method for performing a chemical reaction is provided. The method comprises the following steps: (a) Inserting one end of the elongate member into the second end of the tube assembly such that the one end of the elongate member extends into the first chamber of the tube assembly; (b) Decompressing a spring located at a second end of the tube assembly by unlocking a spring retainer of the tube assembly; and (c) piercing the membrane separating the first chamber of the tube assembly from the second chamber of the tube assembly such that one end of the elongate member extends into the second chamber of the tube assembly.

According to some embodiments of the present disclosure, a method for performing a chemical reaction is provided. The method comprises the following steps: (a) Inserting a swab into a first chamber of a testing device, the first chamber containing a first fluid mixture; (b) Decompressing a first spring located in a first spring chamber of the test apparatus, the first spring decompressing causing a fluid mixture in the first chamber to flow into the first spring chamber of the test apparatus, the fluid mixture being filtered through silica en route to the first spring chamber; and (c) decompressing a second spring located in a second spring chamber of the test apparatus, the second spring decompressing causing a second fluid in the second spring chamber of the test apparatus to filter through silica on way to the second chamber.

The above summary is not intended to represent each embodiment, or every aspect, of the present disclosure. Other features and benefits of the present disclosure will be apparent from the detailed description and drawings that follow.

Drawings

At least one of the figures of the present patent or application is completed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the office upon request and payment of the necessary fee;

FIG. 1 illustrates an exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 2 illustrates an exploded view of the test apparatus of FIG. 1, according to some embodiments of the present disclosure;

FIG. 3 illustrates a cross-sectional view of an exploded view of the test apparatus of FIG. 1, according to some embodiments of the present disclosure;

FIG. 4 illustrates a cross-sectional view of the test apparatus of FIG. 1 in a first configuration, according to some embodiments of the present disclosure;

FIG. 5 illustrates a cross-sectional view of the test apparatus of FIG. 1 in a second configuration, according to some embodiments of the present disclosure;

FIG. 6-1 illustrates a cross-sectional view of a second exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 6-2 illustrates a cross-sectional view of a third exemplary test apparatus according to some embodiments of the present disclosure;

6-3 illustrate cross-sectional views of a fourth exemplary test apparatus according to some embodiments of the present disclosure;

6-4 illustrate cross-sectional views of a fifth exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 7 illustrates a process for performing a test according to some embodiments of the present disclosure;

fig. 8A illustrates a cross-sectional view of a first exemplary electronic device, in accordance with some embodiments of the present disclosure;

fig. 8B illustrates a cross-sectional view of a second exemplary electronic device, according to some embodiments of the present disclosure;

FIG. 9 illustrates an exemplary modular assembly of an electronic device according to some embodiments of the present disclosure;

FIG. 10 illustrates a sixth exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 11 illustrates a cross-sectional view of the sixth test apparatus of FIG. 10, according to some embodiments of the present disclosure;

FIG. 12A illustrates a cross-sectional view of a seventh exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 12B illustrates a cross-sectional view of a seventh test device in a first position according to some embodiments of the present disclosure;

FIG. 12C illustrates a cross-sectional view of a seventh test device in a second position according to some embodiments of the present disclosure;

FIG. 12D illustrates a cross-sectional view of a seventh test apparatus in a third position according to some embodiments of the present disclosure;

FIG. 12E illustrates a cross-sectional view of a seventh exemplary test device in a fourth position according to some embodiments of the present disclosure;

FIG. 13 illustrates a cross-sectional view of an eighth exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 13-1 illustrates a cross-sectional view of a portion of a ninth exemplary test apparatus according to some embodiments of the present disclosure;

FIG. 14 illustrates a cross-sectional view of a tenth exemplary test apparatus according to some embodiments of the present disclosure; and

fig. 15 illustrates steps of using an exemplary test device according to some embodiments of the present disclosure.

While the disclosure is susceptible to various modifications and alternative forms, specific implementations and embodiments thereof have been shown by way of example in the drawings and will herein be described in detail. It should be understood, however, that the disclosure is not intended to limit the disclosure to the particular forms disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.

Detailed Description

Embodiments of the present disclosure provide a simple, low cost system and method for facilitating biochemical reactions that require multiple sequential compartments or chambers, whether due to different reagents, temperatures, or other requirements. In some embodiments, a mechanism for storing potential energy is described that can be released by a simple mechanical trigger to effect movement of fluid between chambers. For example, the potential energy may be stored as a user compressed spring or pre-compressed spring having a solenoid that triggers the release of spring energy and forces fluid from the first chamber to the second chamber through the injector mechanism.

Various embodiments are described with reference to the drawings, wherein like reference numerals are used to refer to like or equivalent elements throughout. The drawings are not necessarily to scale, but are merely provided to illustrate various aspects and features of the disclosure. Numerous specific details, relationships, and methods are set forth to provide a full understanding of certain aspects and features of the disclosure, although one of ordinary skill in the relevant art will recognize that such aspects and features may be practiced with other relationships or with other methods without one or more of the specific details. In some instances, well-known structures or operations are not shown in detail for purposes of illustration. The various embodiments disclosed herein are not necessarily limited by the order of acts or events shown, as some acts may occur in different orders and/or concurrently with other acts or events. In addition, not all illustrated acts or events are necessarily required to implement certain aspects and features of the present disclosure.

For the purposes of this detailed description, the singular includes the plural and, where appropriate, the plural unless specifically excluded. The word "comprising" means "including but not limited to". In addition, approximating words such as "about," "nearly," "substantially," "approximately," etc. may be used herein to mean "in," "near," "nearly in," "within 3-5%, within acceptable manufacturing tolerances," or any logical combination thereof. Similarly, the term "vertical" or "horizontal" is intended to include "within 3-5% of vertical or horizontal, respectively. Additionally, directional words such as "top," "bottom," "left," "right," "above," and "below" are intended to refer to equivalent directions as shown in the referenced figures; as understood in the context, from one or more subjects or one or more elements referenced, e.g. from the location of the usual use of one or more subjects or one or more elements; or as described herein.

Fig. 1 illustrates a test apparatus 100 according to some embodiments of the invention. The test apparatus 100 may allow for the advancement of a reaction from one chamber of the test apparatus to another chamber of the test apparatus. The test device 100 may be an inexpensive disposable device. The test apparatus 100 may include an elongate member 102 and a tube assembly 104 having a first end 112 and a second end 110. The elongate member 102 may be a swab, a syringe, or the like. The elongate member 102 is configured to be inserted into the tube assembly 104 at the second end 110. In fig. 1, the tube assembly 104 is shown as a pre-assembly of the test apparatus 100.

Fig. 2 illustrates an exploded view of components of test apparatus 100 according to some embodiments of the present disclosure. Specifically, FIG. 2 illustrates various portions of the preassembled tube assembly 104 of FIG. 1. The tube assembly 104 includes a first cylindrical member 202, a spring 204, a spring retainer 206, a first chamber housing 208 defining a first chamber 407 (fig. 4), and a second chamber housing 210 defining a second chamber 409 (fig. 4). Fig. 3 illustrates a cross-sectional view of an exploded view of a test apparatus 100 according to some embodiments of the present disclosure. Each of the elongate member 102, the first cylindrical member 202, the spring retainer 206, the first chamber housing 208, and the second chamber housing 210 may be made of plastic or other materials. Examples of materials include Acrylonitrile Butadiene Styrene (ABS), polypropylene, polycarbonate, nylon, 3D printing materials, and the like. In some embodiments, the component under stress or subject to mechanical creep (e.g., spring retainer 206) may be metallic. Examples of metals include steel, aluminum, and the like. In some embodiments, the spring 204 is typically a steel spring.

The first cylindrical member 202 is a hollow structure and may include key openings 212 along the side walls of the first cylindrical member 202. The key opening 212 may be a hole in the first cylindrical member 202. The axis of the key opening 212 may be perpendicular to the longitudinal axis of the first cylindrical member 202. Although a single key opening 212 is shown for the tube assembly 104, in some embodiments, multiple key openings may be provided consecutively along, for example, the sidewall of the first cylindrical member 202. The key openings 212 provide a catch that can receive the protrusion 316 or other portion of the spring retainer 206 when the tube assembly 104 is preassembled as shown in FIG. 1. The spring retainer 206 may be hollow and substantially cylindrical, with the inner diameter of the spring retainer 206 being smaller than the inner diameter of the first cylindrical member 202 of the tube assembly 104. The spring retainer 206 may fit within the hollow structure of the first cylindrical member 202.

When the protrusion 316 of the spring retainer 206 does not pass through the key opening 212 of the first cylindrical member 202, the spring retainer 206 is unlocked and free to move within the hollow first cylindrical member 202. The spring 204 may urge the spring retainer 206 such that the spring retainer 206 moves along the longitudinal axis of the first cylindrical member 202. When the protrusion 316 of the spring retainer 206 passes through the key opening 212 of the first cylindrical member 202, the spring retainer 206 is locked and thus retains its position relative to the first cylindrical member 202.

When the spring retainer 206 is in the locked position, the spring 204 cannot move the spring retainer 206. In some embodiments, when the spring retainer 206 is in the locked position, the spring 204 is compressed, thereby storing potential energy. That is, when in the locked position, the spring retainer 206 prevents decompression of the spring 204, and when in the unlocked position, allows the spring 204 to decompress and move the spring retainer 206 longitudinally along the first cylindrical member 202.

The spring retainer 206 is in a locked position so as not to move along the tube assembly 104, or the spring retainer 206 is in an unlocked position so as to allow movement along the tube assembly 104, which may affect the position of the elongate member 102. The elongate member 102 is inserted into the second end 110 of the tube assembly 104. The elongate member 102 can have a tip 308 and a handle end 302. The tip 308 of the elongate member 102 is inserted into the second end 110 of the tube assembly 104. The handle end 302 facilitates manual manipulation of the elongate member 102 by a user to position the elongate member 102 within the tube assembly 104.

The elongate member 102 can be configured to include multiple portions between the tip 308 and the handle end 302. In some embodiments, the elongate member 102 includes a cylindrical portion 314, a flared portion 304, and a stop portion 312, the cylindrical portion 314 having a constant radius, the flared portion 304 having a varying radius that increases over the length of the flared portion 304 as the flared portion extends away from the cylindrical portion 314, the stop portion 312 having one or more tabs 306, the tabs 306 for preventing removal of the elongate member 102 from the tube assembly 104 after insertion.

In some embodiments, the elongate member 102 further comprises a ridge 310. The ridge 310 may be used to secure the O-ring to the elongate member 102 such that the O-ring may form a tight fit between the first chamber housing 208 and the elongate member 102 when the tip 308 of the elongate member 102 is inserted into the second end 110 of the tube assembly 104. In some embodiments, the O-ring may be replaced by a component that has an easier manufacturing process (e.g., a "2 shot" process using a thermoplastic elastomer (TPE) or the like) as an integral seal. In some embodiments, the elongate member 102 is configured to mate with the first chamber housing 208 such that very little liquid is lost when liquid is injected into the first chamber.

Fig. 4 illustrates a cross-sectional view of test apparatus 100 in a first configuration, according to some embodiments of the present disclosure. In fig. 4, the tube assembly 104 is preassembled. When the tube assembly 104 is constructed, the first cylindrical member 202 is attached to the first chamber housing 208 of the second chamber housing 210. The spring retainer 206 is in the locked position such that the protrusion 316 of the spring retainer 206 penetrates the key opening 212 of the first cylindrical member 202. The spring 204 compresses and is positioned at the second end 110 of the tube assembly 104. The compressed spring 204 urges the spring retainer 206, but the protrusion 316 prevents the spring retainer 206 from moving away from the second end 110 of the tube assembly 104.

In the first configuration, the elongate member 102 is inserted into the tube assembly 104 such that the tip 308 of the elongate member 102 is positioned within the first chamber 407 defined by the first chamber housing 208. A seal 402 or diaphragm is provided, the seal 402 or diaphragm separating the volume of the first chamber 407 defined by the first chamber housing 208 and the volume of the second chamber 409 defined by the second chamber housing 210. In the first configuration, the seal 402 is not broken by the tip 308 of the elongate member 102. In some embodiments, a seal may be provided on the first chamber housing 208 such that the tip 308 of the elongate member 102 pierces the seal to enter the first chamber 407.

In the first configuration, the elongate member 102 is prevented from being pushed further into the tube assembly 104 by the spring retainer 206. The radius of the elongate member 102 at the portion 406a of the elongate member 102 is comparable to or greater than the inner diameter of the spring retainer 206 such that pushing the elongate member 102 into the tube assembly 104 causes interference between the elongate member 102 and the spring retainer 206 at the portion 406a such that the elongate member 102 cannot pass further relative to the spring retainer 206. The interference is provided here as an example, but other mechanical alternatives may be used to couple the elongate member 102 to the spring retainer 206. In some embodiments, instead of the portion 406a of the elongate member 102, the portion 406b of the elongate member 102 may be provided with a radius comparable to or greater than the inner diameter of the spring retainer 206, such that pushing the elongate member 102 into the tube assembly 104 results in interference that prevents the elongate member 102 from being pushed further relative to the spring retainer 206. In some embodiments, both portions 406a and 406b are disposed on the elongate member 102. Other alternatives may also be used to prevent further urging of the elongate member 102 relative to the spring retainer 206. For example, instead of varying the radius along the elongate member 102 to cause interference, the inner diameter of the spring retainer 206 may be reduced at one end such that the portion 406a of the elongate member 102 is larger than the reduced inner diameter at one end.

The tabs 306 prevent removal of the elongate member 102 from the tube assembly 104 as the interference between the elongate member 102 and the spring retainer 206 prevents the tip 308 of the elongate member 102 from moving further toward the first end 112 of the tube assembly 104. When an attempt is made to remove the elongate member 102 from the tube assembly 104, the tabs 306 are configured to snap onto the spring retainer 206. This serves both to prevent removal of the elongate member 102 and to drive the elongate member 102 forward with the spring retainer 206. Particular embodiments of one or more of the fins 306 may be made in any manner, including the 3D printing configuration shown, a two-component injection moldable assembly that separates at or near the fins (for manufacturing purposes, or for moving the fin mechanism within the spring holder 206 to cause the same unidirectional capture). The first chamber housing 208 includes a flared portion 404, the flared portion 404 allowing the flap 306 to move the cylindrical member 202 further downward when the spring retainer 206 is released and placed in the unlocked position.

Fig. 5 illustrates a cross-sectional view of test apparatus 100 in a second configuration, according to some embodiments of the present disclosure. In the second configuration, the spring retainer 206 is in the unlocked position such that the protrusion 316 no longer penetrates the key opening 212. Thus, the spring 204 can urge the spring retainer 206 along the longitudinal axis of the tube assembly 104 toward the first end 112 of the tube assembly 104. Because the spring retainer 206 is coupled to the elongate member 102 at the portion 406a (fig. 4) of the elongate member 102, the elongate member 102 moves a similar distance along the longitudinal axis of the tube assembly 104 as the spring retainer 206. In some embodiments, the elongate member 102 may be removed from the portion 406a due to manufacturing tolerances, but the tab 306 engages the spring retainer 206 and causes the elongate member 102 to move a similar distance along the longitudinal axis of the tube assembly 104 as the spring retainer 206. In some embodiments, the portion 406b (fig. 4) of the elongate member 102 has a radius that is greater than the inner diameter of the spring retainer 206, and the portion 406b engages the spring retainer 206 when the elongate member 102 moves a similar distance as the spring retainer 206.

Moving the elongate member 102 along the longitudinal axis of the tube assembly 104 toward the first end 112 of the tube assembly 104 causes the tip 308 of the elongate member 102 to pierce the seal 402, which seal 402 separates the volume of the first chamber 407 (fig. 4) from the volume of the second chamber 409. In some embodiments, the seal 402 is pierced to combine the volume of the first chamber and the volume of the second chamber such that the reagent mixes between the two chambers. In some embodiments, a solenoid is used to push the protrusion 316 of the spring retainer 206 to move the protrusion 316 away from the key opening 212, causing a transition from the first configuration shown in fig. 4 to the second configuration shown in fig. 5. When the protrusion 316 is pushed, the spring retainer 206 may deform such that at least a portion of the spring retainer 206 including the protrusion 316 has a deformed cross-section as compared to a portion of the spring retainer 206 that mates with the elongate member 102.

In some embodiments, the test device 100 is a disposable multi-chamber device that is used only once to prevent cross-contamination. The solenoid that triggers the transition from the first configuration to the second configuration may be part of a reusable device, controlled by electronic circuitry, to control timing of reaction advancement (as well as other features such as heating or fluorescence measurement). Although the spring 204 is disposed within the tube assembly 104, in some embodiments, the spring 204 may simply be a mechanism for providing mechanical energy to controllably advance the elongate member 102 along the tube assembly 104. That is, the spring 204 may be part of a repeatable apparatus that houses the solenoid. For example, the spring 204 may include a mechanism using pneumatic or compressed gas, or other potential energy forming mechanism, or a simple electromagnet.

Although a first configuration and a second configuration are shown in fig. 4 and 5, respectively, some embodiments of the present disclosure include a test apparatus having more than two chambers. Such test equipment may be stacked to move fluid between more than two chambers provided, for example, from chamber 1 to chamber 2 by a first trigger, then from chamber 2 to chamber 3 by a second trigger, then from chamber 3 to chamber 4 by a third trigger, and so on. The multiple chambers may be separated by a seal (e.g., seal 402). In some embodiments, the first cylindrical member 202 may be provided with a plurality of key openings (e.g., key openings 212) such that the spring retainer 206 transitions between unlocking and locking to successive key openings toward the first end 112 of the tube assembly 104 after each successive trigger.

In some embodiments, each successive chamber may include a different reagent than an adjacent chamber. For example, the first chamber may include a first reagent and the second chamber may include a second reagent. The first reagent may be different from the second reagent. Similarly, the second chamber and the third chamber may have different reagents such that the second reagent in the second chamber is different from the third reagent in the third chamber. In some embodiments, the third reagent is the same as the first reagent. In some embodiments, rather than each chamber having a different reagent, each chamber is maintained at a different temperature. In some embodiments, different combinations of temperatures may be used in different chambers.

Fig. 6-1 illustrates a cross-sectional view of a test apparatus 600 according to some embodiments of the present disclosure. The test apparatus 600 includes an elongate member 602 and a tube assembly 604. The tube assembly 604 includes a first chamber housing 606 and a second chamber housing 608, wherein at least a portion of the first chamber housing 606 defines a first chamber 607 and the second chamber housing 608 defines a second chamber 609. In fig. 6, the first chamber housing 606 is a continuous portion that provides similar functionality as the combined portion of the first chamber housing 208 (fig. 4) and the first cylindrical member 202 (fig. 4). The tube assembly 604 includes a key opening 612 and a seal 616, the key opening 612 being similar to the key opening 212 (fig. 4), the seal 616 being similar to the seal 402 (fig. 4). The test apparatus 600 is shown in the first configuration because the tip 614 of the elongate member 602 is positioned within the first chamber 607 and does not pierce the seal 616. The tube assembly 604 may include a cap 610. Cap 610 may be removed prior to insertion of elongate member 602 or, in some embodiments, cap 610 includes a membrane that may be pierced by elongate member 602 when tube assembly 604 is inserted.

Fig. 6-2, 6-3, and 6-4 illustrate cross-sectional views of test equipment 620, 640, and 660, respectively, according to some embodiments of the present disclosure. The testing apparatus 620 and 640 are shown in a first configuration in which the tip 634 (654) of the elongate member 622 (642) is positioned within the first chamber 627 (647) of the testing apparatus 620 (640). The second chamber 629 (649) of the test device 620 (640) remains sealed and separated from the first chamber 627 (647). The first chamber 627 (647) is defined by a first chamber housing 625 (645). The test equipment 660 is shown in a second configuration in which the tip 675 of the elongate member 662 is positioned within the second chamber 669 of the test equipment 660.

The test apparatus 100 of fig. 4 has a spring retainer 206, the spring retainer 206 being positioned such that when in the locked position, the spring 204 compresses only a small portion of the spring retainer 206 at one end thereof. In fig. 6-2 through 6-4, test equipment 620, 640, and 660 have spring holders 631, 651, and 671, with spring holders 631, 651, and 671 positioned such that springs 633, 653, and 673 compress substantially the entire spring holders 631, 651, and 671, respectively. When in the locked position, the spring holders 631, 651 and 671 are compressed by the springs 633, 653 and 673 because the protrusions 632, 652 and 672 prevent movement of the spring holders 631, 651 and 671 while the force of the springs 633, 653 and 673 is transferred through the entirety or substantially the entirety of the spring holders 631, 651 and 671.

Test equipment 660 is shown in a second configuration, as compared to test equipment 620 and 640. The test device 660 includes a first key opening 674 and a second key opening 670. The first key opening 674 is similar to the key opening 212 (fig. 4) and allows the protrusion 672 to prevent the spring holder 671 from moving in a direction toward the second chamber 669 of the test equipment 660 when in the first configuration. The second key opening 670 is optional and is provided as a second optional feature to prevent the spring holder 671 from moving further toward the second chamber 669 when in the second configuration.

Fig. 6-4 illustrate the test apparatus 660 in a second configuration having a plurality of guards for preventing further movement of the elongate member 662 when the tip 675 is in the second chamber 669. For example, the shape of the first chamber housing 665 that interfaces with the shape of the elongate member 662 may prevent further movement, and the spring retainer 671 against the first chamber housing 665 may prevent further movement, etc.

The tube assemblies 624, 644, and 664 of the test apparatus 620, 640, 660 have first cylindrical members 626, 646, and 666, respectively, with flanges 623, 643, and 663. Flanges 623, 643, and 663 may help the tube assemblies 624, 644, and 664 to be vertically positioned in the holder, with flanges 623, 643, and 663 being points of contact between the tube assemblies 624, 644, and 664 and the holder. In addition, the second chamber housings 628, 648, and 668 have different shapes and are thinner as compared to, for example, the second chamber housing 210 of fig. 4. For optical reading, tube assemblies 620 and 660 may be preferred, and tube assembly 640 may be used for laboratory standard '650 μl' tubes.

Fig. 7 illustrates a process for performing a test according to some embodiments of the present disclosure. Step 702 shows an electronic device 710, the electronic device 710 having visual indicators and/or buttons 716 and a receptacle 714 for receiving a tube assembly 712. The tube assembly 712 may be similar or identical to the tube assembly 104 of fig. 1. Step 704 shows tube assembly 712 inserted into electronic device 710. Step 706 shows a subject 718 having a swab 720 to collect a sample. The swab 720 is similar or identical to the elongate member 102 of fig. 1. Step 708 shows the swab 720 inserted into the tube assembly 712 to test the collected sample. In some embodiments, the sample is a biological sample obtained from a subject in need of testing, e.g., a human subject. The biological sample may be a biological fluid such as saliva, nasal fluid, mucus. The biological sample may include cells or genetic material (e.g., nucleic acids), bacteria, viruses, fungi, or combinations thereof from the subject.

Fig. 8A illustrates a cross-sectional view of a first electronic device 800, according to some embodiments of the invention. The first electronic device 800 may include a microprocessor 802, one or more heaters (e.g., 95 degrees celsius heater, 60 degrees celsius heater, etc.), and a fluorometer 806 (e.g., a 3-color fluorometer). The one or more heaters may include a first heater 804a, a second heater 804b, or additional heaters. Microprocessor 802 is configured to interpret and provide a visual indicator based on the results provided by fluorometer 806. In some implementations, the first electronic device 800 may be designed for a dual-lumen tube assembly (e.g., the tube assembly 104 shown in fig. 4). The first chamber (e.g., first chamber 407 shown in fig. 4) may be positioned such that a first heater 804a of the first electronic device 800 maintains the first chamber at a first temperature (e.g., 95 degrees celsius) and a second heater 804b of the first electronic device 800 maintains the second chamber (e.g., second chamber 409 shown in fig. 4) at a second temperature (e.g., 60 degrees celsius).

Fig. 8B illustrates a cross-sectional view of a second electronic device 801 according to some embodiments of the invention. The second electronic device 801 is similar to the first electronic device 800, but the second electronic device 801 includes a solenoid 810 that can be used to move a protrusion (e.g., protrusion 316 of fig. 4) to advance an elongate member (e.g., elongate member 102 of fig. 4) and pierce a membrane separating two chambers of a test assembly. The first electronic device 800 (fig. 8A) may be used with a tube assembly that requires an operator to manually advance an elongated member, while the second electronic device 801 (fig. 8B) may be used with a tube assembly that provides movement of a spring holder by a solenoid (e.g., tube assembly 104 shown in fig. 4). Microprocessor 802 can be used to time solenoid 810 in second electronic device 801 to meet more precise timing of when the elongate member is advanced. Fig. 9 illustrates a modular assembly of an electronic device according to some embodiments of the present disclosure. In one example, for batch testing, an electronic device (e.g., electronic device 800 or 801) may be arranged as shown in fig. 9.

Fig. 10 illustrates a test apparatus 1000 according to some embodiments of the present disclosure. The test apparatus 1000 includes an elongate member 1002 and a tube assembly 1004. Fig. 11 illustrates a cross-sectional view of a test apparatus 1000 according to some embodiments of the present disclosure. The elongate member 1002 includes a button 1010, the button 1010 facilitating the operator or user to push the elongate member 1002 into the tube assembly 1004. The elongate member 1002 also includes a plurality of one-way detents that can be clamped to the tube assembly 1004 at different points 1012 as the elongate member 1002 is pushed into the tube assembly 1004. In some embodiments, a clicking sound is generated when the elongate member 1002 is pushed into the tube assembly 1004. The clicking sound may provide a human perceptible indication to the operator that the elongate member 1002 is located in the first chamber of the tube assembly 1004, the second chamber of the tube assembly 1004, the third chamber of the tube assembly 1004, and so on. The human perceptible indication lets the user know the distance that the elongate member 1002 is pushed as the reaction proceeds. In some embodiments, the elongate member 1002 is shaped such that once the brake is reached, more force is required to push the elongate member 1002 to the next brake. This force feedback may also be used to provide an indication to an operator or user of how far the elongate member 1002 has been pushed into the tube assembly 1004. The manually pushed test device 1000 may be used with the electronic device 800 of fig. 8A, while the test device 100 of fig. 4 may be used with the electronic device 801 of fig. 8B. Fig. 11 also shows how the first heater 804a and the second heater 804b will engage the tube assembly 1004.

Although fig. 10 and 11 are described in the context of human pushing, other pushing mechanisms may be used. For example, a force applied from an external mechanism, such as a spring, may be used to push the elongate member 1002 into the tube assembly 1004. In some embodiments, the external mechanism may include a gear motor, a servo, a pneumatic piston, or the like.

Fig. 12A illustrates a cross-sectional view of a test apparatus 1200 according to some embodiments of the invention. Fig. 12B-12E provide different positions of the test apparatus 1200. The test apparatus 1200 includes a tube assembly 1204 and an elongate member 1202. The elongate member 1202 includes a button 1201 and a spring 1203. The tube assembly 1204 includes a first chamber housing 1208 defining a first chamber and a second chamber housing 1210 defining a second chamber. The first chamber housing 1208 also includes a thin portion 1207, and the solenoid can deform the thin portion 1207 to trigger. In fig. 12B, tube assembly 1204 is separated from elongate member 1202. In fig. 12C, the tube assembly 1204 receives the elongate member 1202 until the elongate member 1202 snaps at the retainer portion 1206 of the first chamber housing 1208. The retainer portion 1206 prevents the elongate member 1202 from advancing beyond the first chamber.

In fig. 12D, any further pushing down on button 1201 results in compression of spring 1203 while elongate member 1202 remains within the first chamber. The button 1201 snaps into the stopper of the tube assembly 1204, so that the compression spring 1203 remains compressed. In fig. 12E, a solenoid may be used to deform the thin portion 1207 of the first chamber housing 1208 such that the clamping effect causes the retainer portion 1206 to expand. As shown in fig. 12E, this expansion allows the spring 1203 to advance the elongate member 1202 into the second chamber. Comparing fig. 12D and 12E, the spring 1203 is compressed in fig. 12D and decompressed in fig. 12E.

Fig. 13 illustrates a cross-sectional view of a test apparatus 1300 according to some embodiments of the present disclosure. The test device 1300 includes a swab 1301 inserted into a sample collection and extraction chamber 1302. The swab 1301 includes a tip 1304 containing a sample. The sample and collection chamber 1302 may include a 'washboard' geometry 1303 such that when the tip 1304 of the swab 1301 is inserted into the sample and collection chamber 1302, the washboard geometry 1303 may deform the tip 1304 and massage the tip 1304 to help release the sample into the extraction buffer present in the sample and collection chamber 1302. The washboard geometry 1303 eases further manipulation of the swab 1301 by the user to obtain optimal sample extraction. The extraction buffer present in the sampling and collection chamber 1302 can include a nucleic acid extraction buffer. The nucleic acid extraction buffer may facilitate a chemical reaction to extract nucleic acids from the sample on the tip 1304 of the swab 1301.

Test apparatus 1300 is similar to test apparatus 100 (FIG. 4) in that a spring and a spring retainer are disposed in a hollow tubular member. The test apparatus 1300 includes a first spring 1305 disposed in a first hollow tubular member 1307, the first spring 1305 being shown in fig. 13 as being compressed by a first spring retainer 1306. The first spring retainer 1306 operates in the first hollow tubular member 1307 in a similar manner as described above with respect to the first cylindrical member 202 in fig. 4. When the first spring holder 1306 is released, the first spring 1305 decompresses, pushing the first spring holder 1306 and creating a vacuum in a chamber within the first hollow tubular member 1307, wherein the first spring 1305 is located within said first hollow tubular member 1307. Creating a vacuum within the chamber causes drawing of the sample and extraction buffer mixture in the sample and collection chamber 1302. That is, the higher pressure (e.g., atmospheric pressure) within the sample and collection chamber 1302 pushes the sample and extract buffer mixture due to the vacuum created. The sample and extraction buffer mixture flows from the sample and collection chamber 1302 through the flexible ball 1308, which acts as a check valve. The sample and extraction buffer mixture further flows through a frit (and/or filter) 1312. Although described in the context of creating a vacuum, the release of the first spring 1305 may create a vacuum, or in some embodiments a positive pressure, within the hollow tube member 1307.

The frit (and/or filter) 1312 may hold silica purification beads for filtering particles from the sample and extraction buffer mixture, as the mixture passes through the frit (and/or filter) 1312 under the direction of the vacuum created. The test apparatus 1300 also includes a second tubular member 1311 having a second spring 1309 and a second spring holder 1310. The second spring 1309 and the second spring holder 1310 help to push (or inject) fluid present in the second tubular member 1311 through the silica beads disposed in the frit (and/or filter) 1312. The flexible ball 1308 acts as a check valve, preventing pushed (or injected) fluid from entering the sampling and collection chamber 1302. A post-wash/concentrate reaction chamber 1313 is provided in the test apparatus 1300 to collect fluid.

The components in fig. 13 may be connected in different orientations, sizes, and shapes, with flow in different directions. Fig. 13 shows that a spring/solenoid system can be used to automate flow through and elute from a silica bead or other filter in order to clean and/or concentrate a sample, such as saliva or nasal fluid.

Fig. 13-1 illustrates an aspect of the test apparatus 1300 of fig. 13. The first chamber 1350 includes a buffer that impregnates a swab (e.g., swab 1301 of fig. 13). Buffer is released from the first chamber 1350 into the waste chamber 1352 through capillaries 1363b, 1363e, 1363 f. On the way to the waste chamber 1352, the buffer passes through the silica at location 1362. The silica filters the particles of interest as the buffer passes through. Then, the spring 1360 is released by triggering the spring holder 1358, which spring holder 1358 pushes the stopper 1356. According to some embodiments of the application, the spring retainer 1358 may be released, for example, by a solenoid. The stopper 1356 pushes the fluid contained in the chamber 1354. In some embodiments, the fluid is amplification buffer, and the stopper 1356 pushes the amplification buffer from the chamber 1354 into the lower chamber 1364 through the capillaries 1363a, 1363 c. The amplification buffer also passes through location 1362 to lower chamber 1364 and is thus also filtered during processing. The smaller diameter of capillaries 1363a-1363f allows fluid movement to be directed by negative and positive pressure created in waste chamber 1352 or chamber 1354 because gravity alone is not desired to move fluid easily through narrow capillaries 1363a-1363 f.

In fig. 13-1, in some embodiments, waste chamber 1352 may include a spring and spring retainer arrangement to create a negative pressure for directing buffer through capillaries 1363e, 1363f, and 1363 b. Additionally, when the spring 1360 is released, a positive pressure may be created that pushes the amplification buffer from the chamber 1354 through the capillaries 1363a, 1363f and into the lower chamber 1364. Spring 1360 also creates a negative pressure through capillary 1363d to assist in positive pressure when released. Using fig. 13 as a comparison, the first hollow tube member 1307 can be used as a waste chamber, wherein the first spring 1305 is used to create a negative pressure to draw sample and extraction buffer into the first hollow tube member 1307. The second spring 1309 can be used to create a positive pressure in the second hollow tube member 1311 to push the amplification buffer to the post-clean/concentrate reaction chamber 1313.

Fig. 14 illustrates a cross-sectional view of a test apparatus 1400 according to some embodiments of the invention. Test apparatus 1400 operates similar to test apparatus 1300 of fig. 13. The swab 1401 is inserted into a sample collection and extraction chamber 1402, which sample collection and extraction chamber 1402 may have a washboard geometry for automated swab massaging. The first trigger spring 1405 may be used to draw a vacuum on the sample through silica beads included in the frit and/or filter 1412. Frit and/or filter 1412 is configured to hold silica purification beads. The second spring 1409 may be used to push the elution/reaction buffer through the frit and/or beads in the filter 1412 and into the amplification chamber 1413. The flexible ball 1408 may be configured to act as a check valve. The amplification chamber 1413 serves as a post-clean concentration reaction chamber. The test apparatus 1400 of fig. 14 is housed in a cylindrical design when compared to the test apparatus of fig. 13.

The components may be connected in different orientations, sizes, and shapes with flow in different directions. Similar to fig. 13, fig. 14 shows that a spring/solenoid system can be used to automate flow through and elute from a silica bead or other filter in order to clean and/or concentrate a sample, such as saliva or nasal fluid.

Fig. 15 illustrates steps of using a test apparatus 1500 according to some embodiments of the present disclosure. The test apparatus 1500 is similar to the test apparatus 1400 of fig. 14 and operates similarly to the test apparatus 1400 of fig. 14. Fig. 15, panel (a) shows a first step of eluting the swab, fig. 15, panel (B) shows a second step of activating the first spring, fig. 15, panel (C) shows a third step of applying the sample, fig. 15, panel (D) shows a fourth step of activating the second spring, and fig. 15, panel (E) shows a fifth step of eluting the sample.

In panel (a) of fig. 15, the swab 1501 is inserted into the extraction buffer 1502 in the first chamber of the test device 1500. As shown, lyophilized particles 1503 are disposed in a second chamber of test apparatus 1500. The swab 1501 is immersed in the extraction buffer 1502 for a predetermined amount of time. The test apparatus 1500 includes a plurality of spring chambers. In fig. 15, test apparatus 1500 includes two spring chambers (a first spring chamber 1504a with a first spring 1505a and a second spring chamber 1504b with a second spring 1505 b). Spring chambers 1504a and 1504b include spring retainers 1506a and 1506b, respectively, that retain springs 1505a, 1505b of spring chambers 1504a, 1504b in a pre-compressed state. The spring retainers 1506a, 1506b are similar or identical to the spring retainers discussed above in connection with fig. 4. Also included in the test apparatus 1500 is a filter 1507.

In fig. 15, insert (B), the first spring 1505a is activated. According to some embodiments of the present disclosure, the first spring 1505a is activated in a similar manner as discussed above in connection with the release spring retainer. When activated, first spring 1505a generates a vacuum pressure that draws fluid (e.g., extraction buffer 1502) from the first chamber of test apparatus 1500 into first spring chamber 1504a. The path of fluid flow is indicated by arrow 1508 and when released, first spring 1505a is shown in fig. 15, insert (C). The fluid drawn into the first spring chamber 1504a passes through the filter 1507 such that particles of interest from the swab 1501 are retained, and thus the fluid 1509 generated in the first spring chamber 1504a is waste fluid.

In fig. 15, insert (D), the second spring 1505b is activated. According to some embodiments of the present disclosure, the second spring 1505b is activated in a similar manner as discussed above in connection with the release spring retainer. When activated, second spring 1505b pushes second fluid 1510 from second spring chamber 1504b into a second chamber having lyophilized particles 1503. The second fluid 1510 may be an elution or amplification buffer. Panel (E) of FIG. 15 shows the location of the first and second fluids after the two springs 1505a, 1505B are activated. Mixture 1511 includes lyophilized particles 1503.

In fig. 13, 14 and 15, although described as an amplification buffer, the pushed fluid may be an elution buffer. The amplification buffer may be a nucleic acid amplification reagent, including isothermal nucleic acid amplification reagents. The nucleic acid amplification reagents may include Polymerase Chain Reaction (PCR) reagents, recombinase Polymerase Amplification (RPA) reagents, loop-mediated isothermal amplification (LAMP) reagents, rolling Circle Amplification (RCA) reagents, or Strand Displacement Amplification (SDA) reagents.

In some embodiments, the nucleic acid amplification reagents are lyophilized. In some embodiments, the chemical reaction that occurs in the second chamber (e.g., mixture 1511 in the second chamber of panel (E) of fig. 15) is a nucleic acid amplification reaction. The nucleic acid amplification reaction may be PCR, RPA, LAMP, etc. In some embodiments, the second chamber contains a nucleic acid probe comprising a reporter capable of producing a detectable signal. The nucleic acid probe may comprise a nucleotide sequence substantially complementary to an amplicon of the nucleic acid amplification. The second chamber may comprise an exonuclease. The exonuclease may be a double-strand specific exonuclease having 5 'to 3' exonuclease activity.

In fig. 13, 14 and 15, the sample undergoes steps to obtain a processed sample in a second chamber (e.g., the lower chamber of fig. 15). Rather than simply heating the sample in the first/intermediate chamber, a laboratory standard 'silica-based RNA/DNA purification' protocol was employed to remove impurities. In some embodiments, 90% to 99% or more of non-nucleic acid impurities, such as proteins, sugars, cell debris, and buffer components, are removed. The final nucleic acid concentration may be lower than, and thus equal to or higher than the original concentration, allowing for purification and/or concentration changes. The scheme comprises the following steps: (i) Mixing the sample with a larger volume of buffer in a first chamber; (ii) Binding nucleic acids as the sample/buffer passes through the silica particles and into the waste chamber, the sample/buffer being pulled by the piston/spring; and (iii) driving the elution buffer through the silica and into the lower reaction chamber using a second spring.

In some embodiments, the system may be configured with opposing springs in a single tube to allow pushing and retracting of fluid in the tube. In some embodiments, opposing springs allow the swab to be advanced or retracted from one chamber to another chamber in the tube. Multiple spring retainers may be used in these embodiments, allowing the piston or swab to be returned to the previous position.

FIGS. 13 to 15 use an amplification buffer and an amplification reaction as examples. "amplification" as used herein is defined as the creation of additional copies of a nucleic acid sequence, i.e., for example, an amplicon or amplification product. Methods of amplifying nucleic acid sequences include, but are not limited to: isothermal amplification; polymerase Chain Reaction (PCR) and variants of PCR, such as Rapid Amplification of CDNA Ends (RACE); ligase Chain Reaction (LCR); multiplex RT-PCR; immuno-PCR; SSIPA; real-time RT-qPCR; nanofluidic digital PCR.

Non-limiting examples of isothermal amplification include: recombinase Polymerase Amplification (RPA), loop-mediated isothermal amplification (LAMP), helicase-dependent isothermal DNA amplification (HDA), rolling Circle Amplification (RCA), nucleic Acid Sequence Based Amplification (NASBA), strand Displacement Amplification (SDA), nicking Enzyme Amplification Reaction (NEAR), and Polymerase Spiral Reaction (PSR). See, for example, yan et al Isothermal amplified detection of DNA and RNA, march 2014,Molecular BioSystems 10 (5), DOI:10.1039/c3mb70304e, the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the sample comprises a nucleic acid molecule (e.g., RNA or DNA), virus, bacteria, etc., extracted from the subject. The RNA used herein may be any known type of RNA. For example, the RNA can include messenger RNA, pre-messenger RNA, ribosomal RNA, signal recognition particle RNA, transfer messenger RNA, micronuclear RNA, micronucleolar RNA, smY RNA, microcajal body-specific RNA, guide RNA, ribonuclease P, ribonuclease MRP, Y RNA, telomerase RNA component, splice leader RNA, antisense RNA, cis-natural antisense transcript, CRISPR RNA, long non-coding RNA, microrna, piwi-interacting RNA, small interfering RNA, short hairpin RNA, trans-acting siRNA, repeat-related siRNA, 7SK RNA, enhancer RNA, parasitic RNA, type, retrotransposon, viral genome (e.g., viral RNA), viroid, satellite RNA, or vault RNA. As used herein, DNA may include genomic DNA, mitochondrial DNA, viral DNA, complementary DNA (cDNA), single-stranded DNA, double-stranded DNA, circular DNA, and the like.

In some embodiments, the viral genome is extracted from an RNA virus that is a group III (i.e., double-stranded RNA (dsRNA)) virus. In some embodiments, the group III RNA viruses belong to the family of viruses selected from the group consisting of: hepatitis C virus (Amalgaviridae), bovine virus (Birnaviridae), lentivirus (Chrysoviridae), bursae virus (Cystonoviridae), endoviridae (Endornaviridae), hypoviridae (hypoviriae), megabovine virus (Megabinaviridae), parvoviridae (Partiaviridae), calboviridae (Picobiridae), reoviridae (Reoviridae) (e.g., rotavirus (Rotavirus)), whole virus (Totiviridae), quadriviridae. In some embodiments, the group III RNA virus belongs to the genus Botybirneavir. In some embodiments, the group III RNA virus is a non-allocated species selected from the group consisting of: botrytis porus (Botrytis porri) RNA virus 1), beta leafhopper (Circulifer tenellus) virus1, anthrax filiform (Colletotrichum camelliae filamentous) virus1, melon chlorosis yellow (cucurbst yellow) related virus, sclerotinia sclerotiorum recession (Sclerotinia sclerotiorum debilitation) related virus and Trigonella foenum-graecum virus1 (Spissistilus festinus virus 1).

In some embodiments, the viral genome is extracted from an RNA virus that is a group IV (i.e., positive sense single stranded (ssRNA)) virus. In some embodiments, the group IV RNA viruses belong to the order of the viruses selected from the group consisting of: the order of the viruses (Nidovirales), the order of the picornales (Picornavirales) and the order of the Tymovirales (Tymovirales). In some embodiments, the group IV RNA viruses belong to the family of viruses selected from the group consisting of: arterividae (Arteriviridae), coronaviridae (e.g., coronavirus, SARS-CoV), marine viridae (Mesoniviridae), baculoviridae (Roniviridae), bicistroviridae (Diccistroviridae), infectious malacia viridae (Iflavidae), marine RNA viridae (Marnaviridae), picornaviridae (Picornaviridae) (e.g., polioviruses, rhinoviruses (Rhinoviruses) (common cold viruses), hepatitis A viruses), companion cowpea viridae (Secovariae) (e.g., sub-comoviridae), family a (alphaflexvidae), family b (betaflexvidae), family c (gammaflexvidae), family a (tyrvidae), family a (Alphatetraviridae, alvernaviridae, family Astroviridae (astrovidae), family Astroviridae (barnaviidae), family beet necrotic Flaviviridae (benyvidae), family beet necrotic Flaviviridae (bromovidae), family calicividae (calicividae) (e.g., noroviridae), family c (tetravidae), family c (flavoviridae) (e.g., yellow fever virus, west nile virus, hepatitis c virus, dengue virus, village card virus), family c (hepeviidae), family c (hypovirulence virus), family c (subflaviviridae), family luvidae (luvidae), family c (leividae), family c, and family c acid (leividae) (e.g., leividae), wild Tian Bingdu (Nodavididae), permutotetravidae, solanidae (Potyvididae), sarthroviridae, statovirus, pogaviridae (Togaviridae) (e.g., rubella virus, ross river virus, sindbis virus, sinkia virus), lycopersicon esculentum family (Tombusviridae) and Scolopendra family (Virgaviridae). In some embodiments, the group IV RNA viruses belong to the genus of viruses selected from the group consisting of: bacillariornavirus, dicipivirus, labyrnavirus A, paramyxoviridae (Sequiviridae), bluervrus, citrus aspergilus (Cilevirus), hibiscus viridae (Higreevirus), rubus (Idaeovirus), negevirus, euramiavirus (Oulmiavir), polemovirus, sinaivirus and North bean mosaic virus (Sobemovirus). In some embodiments, the group IV RNA viruses are unassigned species selected from the group consisting of: pea aphid virus (Acyrthosiphon pisum virus), basstrorus, black ford virus, blueberry necrotic ringspot virus, cadiistrorus, southern salad virus (Chara australis virus), extra small virus, medlar chlorosis virus, hepelirus, chastetree tick virus, le bro virus, nediistrorus, ciliate virus 1 (Nesidiocoris tenuis virus 1), nifacirus, nilla virus 1, oscievirus, osedax japonicus RNA virus 1, picalivirus, holselinium virus, white vein fungus virus 1, santeuil virus, secalivirus, solenopsis invicta virus 3, marsupium virus. In some embodiments, the group IV RNA virus is a companion virus selected from the group consisting of: sartoviridae, albetovirus, aumaivirus, papanivirus, virtovirus and chronic bee paralysis virus.

In some embodiments, the viral genome is extracted from an RNA virus that is a group V (i.e., negative sense ssRNA) virus. In some embodiments, the group V RNA viruses belong to a phylum or subgenera selected from the group consisting of: negarnaviricota, haploviricotina and polyplovicotina. In some embodiments, the group V RNA viruses belong to the class of viruses selected from the group consisting of: chunqiuviricetes, ellioviricetes, insthoviricetes, milneviricetes, monjiviricetes and yunchangvicetes. In some embodiments, the group V RNA viruses belong to the order of virales selected from the group consisting of: articulavirales, bunyavirales, goujianvirales, jinchuvirales, mononegavirales (Mononegavirales), muvirales and serpentinales (Serpentovirales). In some embodiments, the group V RNA viruses belong to the family of viruses selected from the group consisting of: the tilapia family (Amnonviridae) (e.g., taastrup virus), the Arenaviridae (Arenaviridae) (e.g., lassa virus), the Serpentis family (Aspiviidae), the Bornaviridae (e.g., bornaviridae), the Chu Bingdu family (Chuviridae), the Cruliviridae, the Fei La family (Feraviridae), the Filoviridae (e.g., ebola virus, ma Erbao virus), the fig mosaic family (Fimoviridae), the Hantaviridae (Hantaviridae), the Mikaduoviridae (Jonviridae), the Mymonaviridae, the inner Loviridae (Naiririidae), the Nyamiviridae, the Orthomyxoviridae (Orthomyxoviride) (e.g., influenza virus), paramyxoviridae (e.g., measles virus, mumps virus, nipah virus, hendra virus, and NDV), panaviridae (peribuyveridae), pharmaaviridae, alboviridae (Phenuiviridae), pneumoviridae (Pneumoviridae) (e.g., RSV and metapneumovirus), qin Bingdu (Qinviridae), rhabdoviridae (e.g., rabies virus), supaviridae, tomato spotted wilt virus (tospovidae), and yenvidae (Yueviridae). In some embodiments, the group V RNA viruses belong to the genus of viruses selected from the group consisting of: anphevirus, arlivirus, chengtivirus, crustavirus, tilapineviridae, wastrivirus and Deltavirus (e.g., hepatitis delta virus).

In some embodiments, the viral genome is extracted from an RNA virus, which is a group VI RNA virus, that includes a virally encoded reverse transcriptase. In some embodiments, the group VI RNA viruses belong to the order of the virales. In some embodiments, the group VI RNA viruses belong to a viral family or subfamily selected from the group consisting of: belpaoviridae, caulioviridae (Caulioviridae), transposable viridae (Metaviridae), pseudoviridae (pseudooviridae), retroviridae (retrovirae) (e.g., retroviruses, e.g., HIV), orthoretroviridae (Orthotrovirinae), and foamy viridae (Spumaretrovirinae). In some embodiments, the group VI RNA viruses belong to the genus of viruses selected from the group consisting of: alpha retrovirus (e.g., avian leukosis virus; rous sarcoma virus), beta retrovirus (e.g., mouse breast cancer virus), bovine foamy virus (e.g., bovine foamy virus), delta retrovirus (e.g., bovine leukemia virus; human T-lymphotropic virus), pentravirus (e.g., micropterus dermacorus), equine foamy virus (e.g., equine foamy virus), feline foamy virus (e.g., feline foamy virus), gamma retrovirus (e.g., murine leukemia virus; feline leukemia virus), lentivirus (e.g., human immunodeficiency virus 1; monkey immunodeficiency virus; feline immunodeficiency virus), raw monkey foamy virus (e.g., simisiapumavirus) and simiismallpox (e.g., eastern chimpanzee foamy virus).

In some embodiments, the viral genome is extracted from an RNA virus selected from the group consisting of influenza virus, human Immunodeficiency Virus (HIV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In some embodiments, the RNA virus is an influenza virus. In some embodiments, the RNA virus is an immunodeficiency virus (HIV). In some embodiments, the RNA virus is Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

In some embodiments, the viral RNA is an RNA molecule produced by a virus having a DNA genome (i.e., a DNA virus). As non-limiting examples, the DNA virus is a group I (dsDNA) virus, a group II (ssDNA) virus, or a group VII (dsDNA-RT) virus.

Although the invention has been illustrated and described with respect to one or more implementations, equivalent alterations and modifications will occur to others skilled in the art upon the reading and understanding of this specification and the annexed drawings. In addition, while a particular feature of the invention may have been disclosed with respect to only one of several implementations, such feature may be combined with one or more other features of the other implementations as may be desired and advantageous for any given or particular application.

One or more elements or aspects or steps from one or more of the following claims 1-53, or any portion thereof, may be combined with one or more elements or aspects or steps from one or more of the other claims 1-53, or any portion or combination thereof, to form one or more additional embodiments and/or claims of the present disclosure.

While various embodiments of the present invention have been described above, it should be understood that they have been presented by way of example only, and not limitation. Various changes may be made to the disclosed embodiments in accordance with the disclosure herein without departing from the spirit or scope of the invention. Thus, the breadth and scope of the present invention should not be limited by any of the above-described embodiments. Rather, the scope of the invention should be defined in accordance with the following claims and their equivalents.

Claims

1. An apparatus, comprising:

an elongated member; and

a tube assembly having a first end and a second end, the tube assembly configured to receive the elongate member at the second end, the tube assembly comprising:

a plurality of chambers including a first chamber and a second chamber, the first chamber and the second chamber separated by a membrane;

A spring located at the second end of the tube assembly;

a spring retainer configured to prevent decompression of the spring when in the locked position and to allow decompression of the spring when in the unlocked position.

2. The apparatus of claim 1, wherein the tube assembly further comprises a key opening for receiving a portion of the spring retainer when the spring retainer is in the locked position.

3. The apparatus of claim 2, wherein the longitudinal axis of the tube assembly and the axis of the key opening are orthogonal.

4. A device according to claim 2 or 3, wherein moving the part of the spring holder away from the key opening places the spring holder in the unlocked position.

5. The apparatus of claim 4, wherein a shape of the spring retainer is deformed when the portion of the spring retainer is removed from the key opening.

6. The apparatus of any one of claims 1-5, wherein the spring is configured to urge the spring retainer toward the first end of the tube assembly when in the unlocked position.

7. The apparatus of claim 6, wherein the elongate member moves toward the first end of the tube assembly when the spring retainer is pushed toward the first end of the tube assembly.

8. The apparatus of claim 6 or 7, wherein the elongate member pierces the membrane separating the first chamber from the second chamber in response to pushing the spring retainer towards the first end of the tube assembly.

9. The apparatus of any one of claims 2 to 8, wherein the spring holder is hollow and substantially cylindrical, the inner diameter of the spring holder being smaller than the inner diameter of the tube assembly.

10. The apparatus of any one of claims 1 to 9, wherein the elongate member has a varying cross-sectional area along a length of the elongate member.

11. The apparatus of any one of claims 1-10, wherein the spring retainer is further configured to secure the elongate member within the tube assembly when the elongate member is received at the second end of the tube assembly.

12. The apparatus of any one of claims 1 to 11, wherein the elongate member comprises a tab configured to prevent removal of the elongate member from the tube assembly.

13. The apparatus of any one of claims 1 to 12, wherein the first chamber comprises a first reagent and the second chamber comprises a second reagent different from the first reagent.

14. The apparatus of any one of claims 1 to 13, wherein the elongate member is a syringe or a swab.

15. The apparatus of any one of claims 1 to 14, wherein a solenoid is used to transition the spring retainer from the locked position to the unlocked position.

16. The device of any one of claims 1 to 15, wherein the spring comprises a compressed gas device or a mechanical spring.

17. The apparatus of any one of claims 1 to 16, wherein the plurality of chambers are arranged in series, the plurality of chambers further comprising at least a third chamber, the third chamber being separate from the first chamber and the second chamber.

18. An assembly comprising a plurality of apparatus according to any one of claims 1 to 17 for processing a plurality of samples, the plurality of apparatus being arranged in an array and configured to process the plurality of samples in parallel or in series.

19. A method of performing a chemical reaction, comprising:

inserting a swab into a first chamber of a testing device, the first chamber comprising a first fluid mixture;

decompressing a first spring located in a first spring chamber of the test apparatus, the first spring decompressing causing a fluid mixture in the first chamber to flow into the first spring chamber of the test apparatus, the fluid mixture being filtered through silica en route to the first spring chamber; and

Decompressing a second spring located in a second spring chamber of the test apparatus, the second spring decompressing causing a second fluid in the second spring chamber of the test apparatus to filter through silica en route to the second chamber.

20. The method of claim 19, wherein decompressing the first spring generates a vacuum pressure and decompressing the second spring generates a positive pressure.

21. The method of claim 19 or 20, wherein the first spring and the second spring decompress simultaneously.

22. The method of claim 19, wherein the first spring decompresses before the second spring decompresses.

23. The method of any one of claims 19 to 22, wherein the swab comprises a biological sample.

24. The method of claim 23, wherein the biological sample comprises saliva, mucus, or nasal fluid.

25. The method of any one of claims 19 to 24, wherein the first fluid mixture comprises an extraction buffer.

26. The method of claim 25, wherein the extraction buffer comprises a nucleic acid extraction buffer.

27. The method of any one of claims 19 to 26, wherein the chemical reaction occurring in the first chamber is nucleic acid extraction.

28. The method of any one of claims 19-27, wherein the second fluid mixture comprises an elution buffer or an amplification buffer.

29. The method of any one of claims 19 to 28, wherein the second chamber contains a nucleic acid amplification reagent.

30. The method of claim 29, wherein the nucleic acid amplification reagent is an isothermal nucleic acid amplification reagent.

31. The method of claim 29 or 30, wherein the nucleic acid amplification reagents comprise Polymerase Chain Reaction (PCR) reagents, recombinase Polymerase Amplification (RPA) reagents, loop-mediated isothermal amplification (LAMP) reagents, rolling Circle Amplification (RCA) reagents, or Strand Displacement Amplification (SDA) reagents.

32. The method of any one of claims 29 to 31, wherein the nucleic acid amplification reagents are lyophilized.

33. The method of any one of claims 19 to 32, wherein the chemical reaction occurring in the second chamber is a nucleic acid amplification reaction.

34. The method of claim 33, wherein the nucleic acid amplification reaction is a Polymerase Chain Reaction (PCR), a Recombinase Polymerase Amplification (RPA), a loop-mediated isothermal amplification (LAMP).

35. The method of any one of claims 19-34, wherein the second chamber comprises a nucleic acid probe comprising a reporter molecule capable of producing a detectable signal, wherein the nucleic acid probe comprises a nucleotide sequence substantially complementary to an amplicon derived from nucleic acid amplification.

36. The method of any one of claims 19-35, wherein the second chamber comprises an exonuclease.

37. The method of claim 36, wherein the exonuclease is a double-strand specific exonuclease having 5 'to 3' exonuclease activity.

38. A method of performing a chemical reaction, comprising:

inserting an end of an elongate member into a second end of a tube assembly such that the end of the elongate member extends into a first chamber of the tube assembly;

decompressing a spring located at the second end of the tube assembly by unlocking a spring retainer of the tube assembly; and

a membrane separating the first chamber of the tube assembly from a second chamber of the tube assembly is pierced such that the one end of the elongate member extends into the second chamber of the tube assembly.

39. The method of claim 38, wherein the one end of the elongate member comprises a biological sample.

40. The method of claim 39, wherein the biological sample comprises saliva or nasal fluid.

41. The method of any one of claims 38 to 40, wherein the first chamber of the tube assembly comprises a first fluid mixture that interacts with the one end of the elongate member, the first fluid mixture comprising an extraction buffer.

42. The method of claim 41, wherein the extraction buffer comprises a nucleic acid extraction buffer.

43. The method of any one of claims 38 to 42, wherein the chemical reaction occurring in the first chamber of the tube assembly is nucleic acid extraction.

44. The method of any one of claims 38 to 43, wherein the second chamber of the tube assembly comprises a second fluid mixture comprising an elution buffer or an amplification buffer.

45. The method of any one of claims 38 to 44, wherein the second chamber of the tube assembly comprises a nucleic acid amplification reagent.

46. The method of claim 45, wherein the nucleic acid amplification reagent is an isothermal nucleic acid amplification reagent.

47. The method of claim 45 or 46, wherein the nucleic acid amplification reagents comprise Polymerase Chain Reaction (PCR) reagents, recombinase Polymerase Amplification (RPA) reagents, loop-mediated isothermal amplification (LAMP) reagents, rolling Circle Amplification (RCA) reagents, or Strand Displacement Amplification (SDA) reagents.

48. The method of any one of claims 45-47, wherein the nucleic acid amplification reagents are lyophilized.

49. The method of any one of claims 38 to 48, wherein the chemical reaction occurring in the second chamber is a nucleic acid amplification reaction.

50. The method of claim 49, wherein the nucleic acid amplification reaction is Polymerase Chain Reaction (PCR), recombinase Polymerase Amplification (RPA), loop-mediated isothermal amplification (LAMP).

51. The method of any one of claims 38 to 50, wherein the second chamber comprises a nucleic acid probe comprising a reporter molecule capable of producing a detectable signal, wherein the nucleic acid probe comprises a nucleotide sequence substantially complementary to an amplicon derived from the nucleic acid amplification.

52. The method of any one of claims 38 to 51, wherein the second chamber comprises an exonuclease.

53. The method of claim 52, wherein the exonuclease is a double-strand specific exonuclease having 5 'to 3' exonuclease activity.