Devices and methods for molecular diagnostic testing

• US 10,112,196 B2
• Filed: 11/15/2017
• Issued: 10/30/2018
• Est. Priority Date: 12/31/2014
• Status: Active Grant

• Current Assignee: Visby Medical, Inc.
• Original Assignee: Click Diagnostics, Inc.
• Inventors: Andreyev, Boris, Chu, Steven, Ciopyk, Brian, De La Zerda, Adam, Huang, Helen, Loney, Gregory, Swenson, David D.
• Primary Examiner(s): Hurst, Jonathan M
• Application Number:
  US15/813,671
• Publication Number:
  US 20180071734A1
• Time in Patent Office: 349 Days
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0605 check valves
• CPC Class Codes: B01L 2400/0611 duck bill valves
• CPC Class Codes: B01L 2400/0644 : rotary valves
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/5029 : using swabs
• CPC Class Codes: B01L 3/527 : for a plurality of reagents
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones

Abstract

A hand-held molecular diagnostic test device includes a housing, an amplification (or PCR) module, and a detection module. The amplification module is configured to receive an input sample, and defines a reaction volume. The amplification module includes a heater such that the amplification module can perform a polymerase chain reaction (PCR) on the input sample. The detection module is configured to receive an output from the amplification module and a reagent formulated to produce a signal that indicates a presence of a target amplicon within the input sample. The amplification module and the detection module are integrated within the housing.

Claims

1. A method of detecting a target nucleic acid using a stand-alone molecular diagnostic test device without any external instrument, comprising:

conveying an input sample from a sample preparation module to an amplification module, the sample preparation module and the amplification module each fixedly coupled within a housing of the stand-alone molecular diagnostic test device, the housing having an overall size of less than about 260 cubic centimeters, the amplification module defining a reaction volume and including a heater;

heating the input sample within at least a portion of the reaction volume via the heater to amplify the target nucleic acid molecule within the input sample thereby producing an output solution containing a target amplicon; and

reacting within a detection module within the housing each of A) the output solution and B) a reagent formulated to produce a signal that indicates a presence of the target amplicon within the output solution, the detection module including a detection surface configured to capture the target amplicon to produce the signal,

the stand-alone molecular diagnostic test device producing the signal in less than about 25 minutes after the input sample is conveyed from the sample preparation module.

2. The method of claim 1, wherein a volume of the output solution produced by the amplification module is at least about 75 μ

l.

3. The method of claim 1, wherein the heating the input sample within the reaction volume includes heating the input sample through at least 30 thermal cycles in a time period of less than 10 minutes.

4. The method of claim 1, wherein the reaction volume is a serpentine flow path, the heater is a first heater, the heating the input sample includes producing a first temperature zone within a first portion of the serpentine flow path via the first heater and producing a second temperature zone within a second portion of the serpentine flow path via a second heater, the method further comprising:

producing within the serpentine flow path a flow of the input sample of at least 0.2 μ

l/sec that repeatedly passes through the first temperature zone and the second temperature zone to heat the input sample through at least 30 cycles.

5. The method of claim 1, wherein the conveying the input sample from the sample preparation module to the amplification module is performed by a fluid pump configured to generate within the housing a force to convey the input sample.

6. The method of claim 5, wherein the conveying into the detection module each of the output solution and the reagent is performed by the fluid pump.

7. The method of claim 1, wherein the stand-alone molecular diagnostic test device consumes a total electrical charge of less than 1200 mAh during the method of detecting the target nucleic acid via the signal.

8. The method of claim 1, further comprising:

heating the input sample, within a lysis module disposed within the housing, to a temperature of about 95 degrees Celsius before the input sample is conveyed to the amplification module.

9. The method of claim 1, further comprising:

heating at least one of the output solution or the reagent within the detection module to facilitate production of the signal.

10. The method of claim 1, wherein the target amplicon is associated with at least one of chlamydia trachomatis, neisseria gonorrhoeae, trichomonas vaginalis, E. coli, Staphylococcus saprophyticus, Enterococcus faecalis, Klebsiella pneumoniae, Proteus, or P. aeruginosa.

11. The method of claim 1, wherein the stand-alone molecular diagnostic test device draws a power of less than about 4000 mW from a power source during the method of detecting the target nucleic acid via the signal.

12. The method of claim 1, wherein the stand-alone molecular diagnostic test device produces the signal in less than about 20 minutes after the input sample is conveyed from the sample preparation module.

13. The method of claim 1, further comprising:

conveying a biological sample into the sample preparation module, the biological sample containing a bodily fluid; and

heating the biological sample within sample preparation module to produce the input sample, the stand-alone molecular diagnostic test device producing the signal in less than about 25 minutes after the biological sample is conveyed into the sample preparation module.

14. The method of claim 13, wherein the bodily fluid includes any one of blood, urine, a male urethral specimen, a vaginal specimen, a cervical swab specimen, or a nasal swab specimen.

15. The method of claim 1, further comprising:

coupling the stand-alone molecular diagnostic test device to a power source.

16. A method of detecting a nucleic acid using a stand-alone molecular diagnostic test device without any external instrument, comprising:

conveying a biological sample into a sample preparation module within the stand-alone molecular diagnostic test device; and

actuating the stand-alone molecular diagnostic test device to cause the stand-alone molecular diagnostic test device to;

heat the biological sample via a heater of a lysis module within the stand-alone molecular diagnostic test device to lyse a portion of the biological sample to produce an input sample;

convey the input sample to an amplification module, the amplification module fixedly coupled within the stand-alone molecular diagnostic test device, the amplification module defining a reaction volume;

heat the input sample within the reaction volume through at least 30 thermal cycles to amplify the nucleic acid molecule within the input sample thereby producing an output solution containing a target amplicon; and

react within a detection module within the stand-alone molecular diagnostic test device each of A) the output solution and B) a reagent formulated to produce a signal that indicates a presence of the target amplicon within the output solution, the detection module including a detection surface configured to capture the target amplicon to produce the signal, the stand-alone molecular diagnostic test device producing, without any external instrument, the signal in less than about 25 minutes after the actuating;

reading a result associated with the signal; and

discarding, after the reading, the stand-alone molecular test device.

17. The method of claim 16, wherein the reading is performed via a detection window defined by a housing of the stand-alone molecular diagnostic test device.

18. The method of claim 16, wherein a volume of the output solution produced by, the amplification module is at least about 75 μ

l.

19. The method of claim 16, wherein:

the heater is a first heater;

the reaction volume is a serpentine flow path; and

the input sample is heated within the reaction volume via a second heater, a first heating portion of the second heater produces a first temperature zone within a first portion of the serpentine flow path and a second heating portion of the second heater produces a second temperature zone with a second portion of the serpentine flow path, the first temperature zone and the second temperature zone maintained such that a flow of the input sample within the serpentine flow path is thermally cycled to amplify the nucleic acid.

20. The method of claim 19, wherein the input sample is heated within the serpentine flow path through at least 30 thermal cycles in a time period of less than 8 minutes.

21. The method of claim 16, wherein:

the actuating the stand-alone molecular diagnostic test device includes manipulating an actuator coupled to the housing, a switch of a control module within the housing being actuated in response to movement of the actuator, the control module providing power from a power source to the first heater when the switch is actuated.

22. The method of claim 16, wherein the conveying the biological sample includes conveying the biological sample into the sample preparation module via an opening defined by a housing of the stand-alone molecular diagnostic test device, at least one of an overall length of the housing or an overall width of the housing being less than about 10.2 centimeters.

23. The method of claim 16, wherein the stand-alone molecular diagnostic test device has a power consumption of less than about 4000 mW from a power source from a time of the actuating until the signal is produced.

24. The method of claim 16, wherein the biological sample includes any one of blood, urine, a male urethral specimen, a vaginal specimen, a cervical swab specimen, or a nasal swab specimen.

25. The method of claim 16, wherein the stand-alone molecular diagnostic device has an overall volume of less than about 260 cubic centimeters.

26. The method of claim 16, further comprising:

coupling, before the actuating, the stand-alone molecular diagnostic test device to a power source.