This application also relates to International Patent Application No. PCT/US2017/032904, internationally filed May 16, 2017 entitled “Flow Control System for Diagnostic Assay System”, which claims priority to U.S. Provisional Patent Application Ser. No. 62/337,446 filed May 17, 2016 entitled “Multi-Chamber Rotating Valve and Cartridge.” Additionally, this application also relates to U.S. patent application Ser. No. 15/157,584 filed May 18, 2016 entitled “Method and System for Sample Preparation”, which is a continuation of U.S. Non-Provisional patent application Ser. No. 14/056,543, filed Oct. 17, 2013, now U.S. Pat. No. 9,347,086, which claims priority to U.S. Provisional Patent Application Ser. No. 61/715,003, filed Oct. 17, 2012, which is a continuation-in-part of U.S. patent application Ser. No. 12/785,856, filed May 24, 2010, now U.S. Pat. No. 8,663,918, which claims priority to U.S. Provisional Patent Application Ser. No. 61/180,494, filed May 22, 2009, and which is also a continuation-in-part of U.S. patent application Ser. No. 12/754,205, filed Apr. 5, 2010, now U.S. Pat. No. 8,716,006, which claims priority to U.S. Provisional Patent Application Ser. No. 61/158,519, filed Apr. 3, 2009. The contents of the aforementioned applications are hereby incorporated by reference in their entirety.
The present invention relates to systems and methods for accelerating Polymerase Chain (PC) reactions, and more particularly to efficiently and effectively heating a PCR chamber by a heating source disposed along a single side of the chamber.
There is continuing interest to improve testing methodologies, facilitate collection and decrease the time associated with clinical laboratories. Particular testing requires that a sample be disrupted to extract nucleic acid molecules such as DNA or RNA.
The number of diagnostic tests performed annually has increased exponentially in the past decade. The use of molecular diagnostics and gene sequencing in research and medical diagnostics is also rapidly growing. For example, DNA testing has also exploded in view of the growing interest in establishing and tracking the medical history and/or ancestry of a family. Many, if not all of these assays, could benefit from a rapid sample preparation process that is easy to use, requires no operator intervention, is cost effective and is sensitive to a small sample size.
Sample collection and preparation is a major cost component of conducting real-time Polymerase Chain Reaction (PCR), gene sequencing and hybridization testing. In addition to cost, delays can lead to the spread of infectious diseases, where time is a critical component to its containment/abatement. In addition to delaying the test results, such activities divert much-needed skilled resources from the laboratory to the lower-skilled activities associated with proper collection, storage and delivery.
For example, a portable molecular diagnostic system could be operated by minimally trained personnel (such as described in US 2014/0099646 A1) and have value with regard to disease surveillance. However, the adoption of such portable systems can be limited/constrained by current methods of sample collection, which require trained personnel to permit safe and effective handling of blood/food/biological samples for analysis. Other limitations relate to: (i) the ability of injected/withdrawn fluids to properly flow, (ii) manufacturability, (iii) cross-contamination of assay fluids which may influence the veracity of test results, (iv) proper admixture of assay fluids to produce reliable test results, and (v) the ability or inability to introduce catalysts to speed the time of reaction,
A need, therefore, exists for an improved disposable cartridge for use in combination with a portable molecular diagnostic/assay system which facilitates/enables the use of minimally-trained personnel, hands-off operation (once initiated), repeatable/reliable test results across multiple assay samples (e.g., blood, food, other biological samples) and an ability to cost effectively manufacture the disposable cartridge for the diagnostic assay system.
The present invention is directed to an system for performing diagnostic testing of an assay fluid. The diagnostic assay system includes a platform configured to receive a disposable cartridge having a sample chamber for receipt of the assay fluid, and an a PCR chamber disposed in fluid communication with the sample chamber for performing target amplification of the assay fluid. A heating source is disposed adjacent a heat exchange surface disposed along at least one side of the PCR chamber and is configured to conform to the contour of the heat exchange surface to accelerate target amplification of the assay fluid. The heating source introduces heat into the assay fluid from one side of the disposable cartridge and, in one embodiment, employs a conformal material interposing the heating source and the heat exchange surface to mitigate the formation of air pockets therebetween.
The present invention is disclosed with reference to the accompanying drawings, wherein:
Corresponding reference characters indicate corresponding parts throughout the several views. The examples set out herein illustrate several embodiments of the invention but should not be construed as limiting the scope of the invention in any manner.
A disposable cartridge is described for use in a portable/automated assay system such as that described in commonly-owned, co-pending U.S. patent application Ser. No. 15/157,584 filed May 18, 2016 entitled “Method and System for Sample Preparation” which is hereby included by reference in its entirety. While the principal utility for the disposable cartridge includes DNA testing, the disposable cartridge may be used to detect any of a variety of diseases which may be found in either a blood, food or biological specimen. For example, blood diagnostic cartridges may be dedicated cartridges useful for detecting hepatitis, autoimmune deficiency syndrome (AIDS/HIV), diabetes, leukemia, graves, lupus, multiple myeloma, etc., just naming a small fraction of the various blood borne diseases that the portable/automated assay system may be configured to detect. Food diagnostic cartridges may be used to detect salmonella, e-coli, staphylococcus aureus or dysentery. Blood diagnostic cartridges may be dedicated cartridges useful for detecting insect or animal borne diseases including malaria, encephalitis and the West Nile virus.
More specifically, and referring to
The disposable cartridge 20 provides an automated process for preparing the fluid sample for analysis and/or performing the fluid sample analysis. The sample preparation process allows for disruption of cells, sizing of DNA and RNA, and concentration/clean-up of the material for analysis. More specifically, the sample preparation process of the instant disclosure prepares fragments of DNA and RNA in a size range of between about 100 and 10,000 base pairs. The chambers can be used to deliver the reagents necessary for end-repair and kinase treatment. Enzymes may be stored dry and rehydrated in the disposable cartridge, or added to the disposable cartridge, just prior to use. The implementation of a rotary actuator allows for a single plunger to draw and dispense fluid samples without the need for a complex system of valves to open and close at various times. This greatly reduces potential for leaks and failure of the device compared to conventional systems. It will also be appreciated that the system greatly diminishes the potential for human error.
In
Depending upon the specific function of the cartridge 20, one important feature of the channels 40, 42 is to facilitate and augment amplification by forming a region which may be heated from the underside of the cartridge 20. During development of the disposable cartridge and diagnostic assay system, the inventors were faced with various challenges associated with accelerating amplification. More specifically, the inventors learned that the use of conventional conductive grease along the mating interface of a channel 42 was inadequate to reach a desired temperature set point, i.e., to transfer heat, within a reasonable time frame. It was at this point that the inventors began conducting a variety of inventive methods and configurations which would lead to a two-fold increase in amplification time. These tests/inventive discoveries are discussed in the subsequent paragraphs.
In
More specifically, the mounting platform 104 includes a circular disc 110 disposed at the center of a rectangular or square mounting plate 112. The circular disc 110 is adjacent to and is contiguous with the underside surface 44S (best seen in
In the described embodiment, the heat source 106 is integrated with the circular disc 106 of the mounting platform 104. The heat source 106 may be any resistive heater, however, in the disclosed embodiment, a low wattage RF heat source or inductive heater may be employed. That is, inasmuch as the diagnostic assay tester 10 is portable, a source of high current may not be readily available. In view of these contingencies, an RF and/or inductive heater may be preferable inasmuch as such heat sources may operate on 6-12 volt battery power. A typical RF heating device may include any strip of material which is responsive to RF energy. Such materials include a molecular lattice which is excited, i.e., vibrates, in the presence of an RF energy field within a particular frequency band.
In
In
With respect to the former, the conformal material 132 is configured to elongate from between about twenty (20%) to about fifty percent (50%) of an original dimension. For example, a conformal material having an original dimension of about 0.5 inches may deform elastically under a tensile load (i.e., pulling the material apart) to between about 0.6 inches to about 0.75 inches. With respect to the latter, it will be appreciated that a conformal material having a Shore-A hardness of between about thirty (30) to about seventy (70) less than about 70 is useful for practicing the inventive features of this disclosure.
Testing of the various configurations described herein provides nearly a two-fold increase in temperature response and accuracy. For most of the assay fluid procedures, temperatures can be controlled to within one degree Celsius (1°). In one embodiment, a thermocouple 136 may be introduced to measure the temperature within the amplification region AR while another thermocouple 138 reads an ambient temperature to establish a baseline or threshold temperature. The thermocouple 136 in the amplification region AR issues an actual temperature signal indicative of an instantaneous temperature of the assay fluid XX. The signal processor 140 is responsive to the actual temperature signal, compares it to a stored threshold temperature signal, and controls the heat source such that the actual temperature is maintained within a threshold range of the threshold temperature. Alternatively, a second thermocouple 138 issues a baseline or ambient temperature signal for comparison to the actual temperature signal. While the illustrated embodiment depicts a thermocouple along the underside surface of the disposable cartridge 20, it will be appreciated that one or both of the thermocouples 136, 138 may be disposed in combination with the contact plate 112, proximal the heat source 106 and juxtaposed the underside of the cartridge rotor 18.
In one embodiment, the conformal coating is disposed over the heating source. This could be some type of elastomer, silicone, foam, epoxy, phase change material, or gel pack. The properties of the material would be such that repeated contact would have minimal effect on its physical integrity (slow wear). This could be done using slip coatings, slip additives or other fillers. Naturally, the conformal properties would be retained. Alternately, the material may be considered a consumable and replaced after a particular lifetime. This would relax the wear tolerance. While the material would not require thermal conductive properties, it is desired. Materials with a low thermal conductivity would likely require thinner coatings to reduce heat transfer times. With the heating element coated with a conformal and thermally conductive film, the reagent vessel can be put in contact. The conformal nature of the film improves surface to surface contact while minimizing small voids that may occur The temperature can then be cycled.
In another embodiment, the wear and damage incurred are abated by actuating the heating element itself. When not being used, the heating element or heat source is retracted away from the heat transfer surface preventing any contact induced damage. When the process calls for heating or cooling, the heating element or vessel is actuated and the two surfaces are pressed together. The conformal and conductive nature of the heating element surface allows for enhanced surface contact and thermal transfer between the heating element and the reagent vessel. The process of actuating the parts can be done a variety of ways depending upon design requirements.
In another embodiment, the heating element is coated with a conformal and preferably thermally conductive coating. In addition, the heating element is spring loaded. This provided partial wear relief if the vessel and heating element are moved while in contact.
In another embodiment, a material having a high coefficient of thermal expansion is employed while also having a conformal characteristic. The material is coated over the heating element. Under non-processing conditions, the coating would not be in contact with the PC chamber or heat transfer interface. Upon heating, the material expands to fill any voids which may exist between the two surfaces. The enhanced surface contact would allow improved thermal transfer. When processing is complete, the material cools and retracts from the vessel surface to allow free movement therebetween.
While the invention has been described with reference to particular embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the scope of the invention.
Therefore, it is intended that the invention not be limited to the particular embodiments disclosed as the best mode contemplated for carrying out this invention, but that the invention will include all embodiments falling within the scope and spirit of the appended claims.
While the invention has been described with reference to particular embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from the scope of the invention.
This application is a Continuation-In-Part of U.S. patent application Ser. No. 16/303,441 entitled “System and Method for Optimizing Heat Transfer for Target Amplification within a Diagnostic Assay System” which claims priority to U.S. Provisional Patent Application Ser. No. 62/344,711, filed Jun. 2, 2016 entitled “Multi-chamber Rotating Valve and Thermal Control In A Microfluidic Chamber”. The contents of the aforementioned applications are hereby incorporated by reference in their entirety. The contents of the aforementioned applications are hereby incorporated by reference in their entirety. The contents of the aforementioned applications are hereby incorporated by reference in their entirety.
Number | Date | Country | |
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62344711 | Jun 2016 | US |
Number | Date | Country | |
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Parent | 16303441 | US | |
Child | 16206253 | US |