The present invention relates to a disposable cartridge for a portable diagnostic assay device, and more particularly, to a system and method for heat optimization of reactions in a portable diagnostic assay system.
Fluid analysis of biological samples such as blood and food samples for assay testing general require a series of process steps. These steps generally require that particular fluids contact a reaction area at different times and in varying secession. Furthermore, each fluid may require different pre-treatment prior to contacting the reaction area such as chemical, optical, thermal, mechanical, magnetic or acoustical pre-treatment. A single fluid sample may be subjected to a variety of steps prior to contact with a reaction area such as heating or ultrasonic processing. As the number of fluids and pre-treatment steps increase, the fluid delivery system becomes more complex.
One of the more recent developments in the field of diagnostic testing relates to a portable diagnostic assay device capable of performing a variety of common and complex laboratory procedures without the requirement for a staff of highly-skilled technicians to perform these procedures in a costly laboratory environment/setting. The portable diagnostic assay device and related diagnostic cartridges are disclosed in a portfolio of issued and pending US and foreign patents/patent applications assigned to Integrated Nano-Technologies located in the town of Henrietta, state of New York, USA. The portable diagnostic assay device comprises a small base unit, i.e., generally smaller than a standard briefcase, for accepting one of many distinct, dedicated, and disposable cartridges prepared for conducting a single assay test. For example, the disposable cartridges may be prepared for testing blood borne diseases, food borne bacteria, and/or animal/insect carrying bacteria and viruses.
The diagnostic cartridges comprise a plurality of chambers each containing a reagent used in the assay test, e.g., PCR primers, enzymes and certain chemical compounds. One method to significantly improve the efficiency and yield of PCR amplification is to heat the reaction at various stages in the assay fluid process. The more rapidly an assay fluid reaches a desired temperature, the more efficient is the process. Furthermore, as the accuracy of the temperature improves, the assay sample yield increases which can reduce the number of cycles required to reach a desired level of PCR amplification.
Of the many variables which can impact temperature, the most dominant is the insulating effects of air trapped between assay fluids and the heat source of the diagnostic assay system. This problem is typically resolved by the use of a conductive cream or gel disposed between the heat source and the assay fluid. While this solution may be satisfactory for machines which typically employ greased bearings and gears, it is not well-suited for a portable laboratory requiring the equivalent of a clean-room environment. Furthermore, the portable diagnostic system employs sensitive electronic circuit boards which can be short-circuited should the conductive cream or gel flow into or across the soldered leads of such PC boards. Finally, the conductive cream or gel must be cleaned with each use and, as such, is not practical for most high cycle machines.
A need, therefore, exists for an efficient, reliable, and practical system and method for optimizing the heat transfer associated with target amplification in a portable diagnostic assay system.
In one embodiment, a method is provided for optimizing the heat transfer when performing target amplification of an assay fluid, comprising the steps of: (i) moving assay fluid through at least one channel disposed along an underside surface of a disposable cartridge of a diagnostic assay test device such that the fluid collects in an amplification region of the channel; (ii) heating the amplification region of the assay channel to heat the assay fluid; (iii) interposing a conformal material between the underside surface of a disposable cartridge and the RF heater, and (iv) applying a contact pressure between underside surface of a disposable cartridge and the RF heater.
In another embodiment, a diagnostic assay system is provided including a mounting platform receiving a disposable cartridge, a heat source disposed in combination with the mounting platform and a multi-axis actuation system operative to rotationally index the cartridge rotor relative to the cartridge body and apply a contact force along a mating interface disposed between the underside surface of the cartridge and the heat source.
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.
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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
In the described embodiment, the heat source 106 is integrated within the circular disc 106 of the mounting plate 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.
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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.
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.
This application 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. 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.
Filing Document | Filing Date | Country | Kind |
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PCT/US2017/035682 | 6/2/2017 | WO | 00 |
Number | Date | Country | |
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62344711 | Jun 2016 | US |