US-12623227-B2 - Apparatus and methods for molecular diagnostics

Abstract

This disclosure relates to apparatus and methods for molecular diagnostics. Certain embodiments include a piston cycled from a first position proximal to a first end of a housing, to a second position proximal to a second end of the housing, and back to the first position proximal to the first end of the housing. In some embodiments, the present disclosure relates to devices, methods, and systems for molecular diagnostics that do not comprise a piston.

Inventors

• Richard Crockett
• Alan Blake
• Michael Karberg
• John Lupher
• CHRIS CLEVER NEE
• Matthew Flint Kepler

Assignees

• NUCLEIN, LLC

Dates

Publication Date

20260512

Application Date

20210304

Claims (20)

1. 1 . An apparatus for performing molecular diagnostics, the apparatus comprising: a chamber comprising: a piston; and a fluid configured to amplify a nucleic acid via thermal cycling; a plurality of reservoirs; a plurality of reaction components, wherein each reaction component is contained in a reservoir of the plurality of reservoirs; and a control circuit configured to receive a first signal from a user interface and transmit a second signal to the array of reaction component, wherein the second signal causes the release of a reaction component from a reservoir into the chamber.
2. 2 . An apparatus according to claim 1 wherein the plurality of reservoirs is arranged in an array.
3. 3 . An apparatus according to claim 1 , wherein the user interface is incorporated into a separate device that transmits the first signal to the control circuit, and wherein the separate device is a smart phone, tablet or computer.
4. 4 . An apparatus according to claim 1 , wherein the reaction component from the array of reaction components is a primer set is configured to replicate a nucleic acid and identify a particular analyte.
5. 5 . An apparatus according to claim 1 , wherein the piston is configured to reciprocate in the chamber, and wherein the apparatus further comprises a light source configured to illuminate the reaction components chamber.
6. 6 . An apparatus according to claim 1 , wherein: the chamber comprises a first end and a second; the apparatus comprises a first heat source proximal to the first end of the chamber.
7. 7 . An apparatus according to claim 6 , further comprising an actuator configured to move the piston in a cycle from a first position proximal to the first end of the chamber, to a second position proximal to the second end of the chamber, and back to the first position proximal to the first end of the chamber.
8. 8 . An apparatus as in claim 7 wherein: the piston is configured to direct the fluid to the second end of the chamber when the piston is in the first position; and the piston is configured to direct the fluid to the first end of the chamber when the piston is in the second position.
9. 9 . An apparatus according to claim 7 , further comprising a controller configured to control the actuator.
10. 10 . An apparatus of claim 9 wherein: the apparatus further comprises a second heat source proximal to a second end of the chamber; the controller is configured to control the first heat source and the second heat source; the controller is configured to control the first heat source to heat the fluid proximal to the first end of the chamber to a temperature between about 92 and 98 degrees Celsius; the controller is configured to control the second heat source to heat the fluid proximal to the second end of the chamber to a temperature between about 60 and 65 degrees Celsius; and the controller is configured to cycle the piston from the first position to the second position and back to the first position between about 30 and 50 cycles.
11. 11 . An apparatus according to claim 10 , wherein: the nucleic acid is DNA or RNA; and the fluid comprises polymerase chain reaction (PCR) reagents or reverse transcriptase polymerase chain reaction (RT-PCR) reagents.
12. 12 . An apparatus according to claim 1 , further comprising an illumination module configured to illuminate an analyte contained in the fluid and a detection module configured to detect a response from the analyte contained in the fluid.
13. 13 . An apparatus according to claim 12 , wherein the detection module detects a response from the analyte in real time during amplification.
14. 14 . An apparatus according to claim 12 , wherein the detection module detects a response from the analyte within an amplification cycle.
15. 15 . An apparatus according to claim 1 , wherein the apparatus weighs less than two pounds.
16. 16 . An apparatus according to claim 1 , wherein the apparatus has two or fewer moving parts.
17. 17 . An apparatus according to claim 1 , wherein the apparatus is powered by a battery having a capacity not greater than 3,000 mAh.
18. 18 . An apparatus according to claim 1 , wherein the apparatus has an operating noise level of less than 10 decibels.
19. 19 . An apparatus according to claim 1 , wherein chamber has a volume of less than 50 microliters.
20. 20 . A method of analyzing a biological sample, the method comprising: obtaining an apparatus for performing molecular diagnostics, where the apparatus comprises: a chamber comprising a piston and a fluid, wherein the fluid is configured to amplify a nucleic acid via thermal cycling; a plurality of reservoirs; a plurality of reaction components, wherein each reaction component is sealed in a reservoir of the plurality of reservoirs; and a control circuit configured to receive a first signal from a user interface and transmit a second signal, wherein the second signal causes the release of a desired reaction component of the plurality of reaction components from a reservoir into the chamber; and selecting the desired reaction component of the plurality of reaction components via the user interface.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a national phase application under 35 U.S.C. § 371 of International Application No. PCT/US2021/020786, filed Mar. 4, 2021, which claims priority to U.S. Provisional Patent Application Ser. No. 62/987,185 filed Mar. 9, 2020, the entire contents of each of which are incorporated herein by reference. BACKGROUND A. Field This disclosure relates to apparatus and methods for molecular diagnostics. More particularly, this disclosure relates to performing molecular diagnostics via a portable device that can provide point-of-care diagnostics. B. Related Art Molecular diagnostics can provide many benefits including early detection of diseases, disorders, or other genetic health-related conditions. Many molecular diagnostic techniques are based on the detection and identification of specific nucleic acids, both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), extracted and amplified from a biological specimen (e.g. blood, saliva or other substances as disclosed herein). Accordingly, while molecular diagnostics provide many benefits, typical molecular diagnostic devices are complicated, expensive, non-portable and require additional equipment and technical expertise for sample preparation, analysis, etc. Despite barriers associated with typical devices, molecular diagnostic tests have the potential to improve health care services, enhance patient outcomes and individualized patient care. In view of the above, there is a need for molecular diagnostics provided by a stand-alone, inexpensive, simple-to-use, portable device suitable for point-of-care use. SUMMARY Briefly, the present disclosure provides devices, methods, and systems for molecular diagnostics comprising a piston that moves between two or more positions to cycle a fluid between two or more temperatures. In some embodiments, the present disclosure provides devices, methods, and systems for molecular diagnostics that do not comprise a piston. Certain embodiments comprise an apparatus for performing molecular diagnostics, where the apparatus comprises: a housing comprising a first end and a second end; a piston disposed within the housing; a first heat source disposed proximal to the first end of the housing; and an actuator configured to move the piston in a cycle between a first position and a second position proximal to the second end of the housing, and back to the first position proximal to the first end of the housing. In some embodiments, the actuator may, after cycling the piston multiple times between the first and second positions, stop the piston at either the first position or the second position when the cycling process is completed. In particular embodiments, at least a portion of the fluid moves in an opposite direction to the piston during the cycle. In certain embodiments the detection module detects a response from the analyte in real time during amplification. In particular embodiments the detection module detects a response from the analyte within an amplification cycle. In some embodiments the amplification cycle has a variable length. Some embodiments further comprise a second heat source proximal to the second end of the housing. In specific embodiments, the first heat source comprises a first heating coil configured to increase the temperature of the first end of the housing when electric current is applied to the first heating coil; and the second heat source comprises a second heating coil configured to increase the temperature of the second end of the housing when electric current is applied to the second heating coil. Certain embodiments further comprise a reaction chamber insert or cartridge configured to be inserted into the housing. In particular embodiments, the reaction chamber insert contains a reaction fluid during use. In some embodiments, the reaction fluid does not contact the piston and the housing during use (for example, when a liner, insert, or other intermediary material is used). In specific embodiments, the housing comprises a fluid configured to replicate (i.e., amplify) a nucleic acid sequence or sequences via thermal cycling. In certain embodiments, the nucleic acid is DNA or RNA or XNA. In particular embodiments, the piston is configured to direct at least a portion of the fluid to the second end of the housing when the piston is in the first position; and the piston is configured to direct at least a portion of the fluid to the first end of the housing when the piston is in the second position. In some embodiments, the fluid comprises reagents for amplification or replication (e.g., via qualitative, quantitative, or semi-quantitative PCR, RT-PCR or other thermal cycling or isothermal techniques). In some embodiments, the fluid comprises polymerase chain reaction (PCR) reagents, such as reagents for reverse transcriptase polymerase chain reaction (RT-PCR), multiplex PCR, nested PCR, asymmetric PCR, hot-start PCR, methylation-specific PCR, allele-s