HK1231115A1 - Cartridge and flow-through apparatus - Google Patents

Description

Box type testing device and through type detection equipment

The present application is a divisional application of a patent application having an application number of 201180055560.1, an application date of 2011, 11/17, and an invention name of "pass-through detection device".

Technical Field

The present invention relates to a cartridge-type test apparatus and a pass-through detection device, and more particularly to a pass-through detection device having a reaction chamber for flow-through hybridization. The invention also relates to an accessory or consumable for use in the diversion hybridization.

Background

The identification or detection of nucleic acid molecules, such as DNA, RNA, antigens, antibodies, proteins, receptors, cells, etc., provides useful information for clinical, diagnostic, and medical applications. For example, genotyping provides useful information about a genetic disease or genetic disorder for reference by a medical professional. However, most identification or detection processes involving nucleic acid molecules require precisely controlled reaction conditions to provide an appropriate and optimal reaction for the particular molecular interaction to occur, such that accurate, predictable and/or consistent results are obtained.

For example, where one process involves the formation of a double-stranded helix from complementary single-stranded molecules in an intermediate process, very precise temperature control is required to be able to distinguish between complete (i.e. 100%) matching sequence homologies of single base variations. In the case of annealing at low temperature, nonspecific binding becomes apparent, and incomplete pairing of low homologous sequence fragments occurs due to the formation of hybrid helices, which increases false detection.

Nucleic acid hybridization techniques have been increasingly used in the detection of nucleic acid molecules for molecular diagnostic applications, and this technique builds on the ability of nucleic acid strands of complementary sequence to form double-helix hybrids.

The conventional reverse dot blot hybridization process for nucleic acids is performed in glass test tubes or plastic bags in a temperature controlled hybridization chamber, and takes hours or even days to complete.

The flow-through nucleic acid hybridization methods described in U.S. patent 6,638,760 and WO2007/115491 are an improvement over conventional processing. In this improved method, a sample solution containing the target analyte is applied to a porous matrix permeable membrane on which capture polynucleotides capable of hybridizing to the target nucleic acid have been immobilized. As the sample solution flows through the porous matrix, the target nucleic acid is captured and detected.

Flow-through hybridization techniques are attractive because they can produce significantly improved signal intensity in a very short time with reduced processing time and the use of small amounts of reagents. In addition, using this method, results can be captured using low-cost trans-membrane-based macroarrays (macro-arrays).

Disclosure of Invention

The present invention provides a cartridge test device adapted for use with a flow-through hybridization detection apparatus having a reaction chamber and a temperature-regulated reaction chamber bottom surface, said cartridge test device comprising a molded main cartridge body, a permeant membrane attached to a downstream side bottom surface of the main cartridge body and adapted to hybridize with passing analytes to capture and display a specific or predetermined sequence of a target analyte, and a sealing means disposed on the downstream side of the permeant membrane, wherein said main cartridge body forms at least one test compartment, said sealing means adapted to cooperate with said reaction chamber bottom surface when used with said flow-through hybridization detection apparatus to provide a hermetic seal on said cartridge test device bottom surface to prevent analyte flow from said test compartment to an adjacent test compartment.

The main cassette comprises a grid constituting a plurality of test units or test compartments adapted to perform hybridization, said sealing means being arranged at a downstream side of said grid to provide a respective liquid seal to each test unit or test compartment to avoid cross-flow contamination between adjacent compartments or to prevent analyte flow from one test unit or test compartment to the other.

The invention provides a through type detection device suitable for flow-through hybridization. In one embodiment, the pass-through detection device is suitable for desktop use. In one embodiment, the reaction chamber is re-closable and is adapted to receive one or a selected plurality of modular cartridge hybridization test devices.

In one aspect, there is provided a flow-through assay device comprising a thermally regulated reaction chamber and an air flow means, wherein the reaction chamber comprises a temperature regulation surface (or reaction surface) adapted to effect flow-through hybridization under thermally regulated and controlled conditions and a thermal regulation component comprising an active electro-thermal regulation means adapted to heat or cool the temperature regulation surface by conduction; and wherein, on the bottom surface of the reaction chamber, a plurality of discharge holes adapted to discharge an excessive or residual liquid within the reaction chamber are distributed.

By distributing a plurality of discharge openings over the reaction surface, it is advantageous to make the suction force on the reaction surface uniform. At the same time, the distributed vent holes allow the reaction chamber to be partitioned into multiple smaller chamber sections to cooperate with a sealed grid formed on the modular cartridge testing device. In this regard, one or a selected plurality of modular cartridge hybridization test devices can be used as required. For example, one or more modular cartridge hybridization test devices may be used, each of which may include a plurality of test units or compartments, depending on the needs.

A suction and discharge net adapted to suck off excess or residual liquid on the bottom surface of the reaction chamber may be connected to the suction opening, the suction and discharge net being adapted to provide an equalized or homogenized suction force or volumetric suction rate to the plurality of discharge openings. It should be noted that uniform suction facilitates a controlled environment that is advantageous for flow-through hybridization.

The suction discharge net may include a plurality of large tubes each connected with the plurality of suction holes via a corresponding plurality of small tubes; and wherein each large tube and the plurality of small tubes connected to the large tube are arranged such that a suction force or a volumetric suction rate exhibited at a suction orifice connected to the plurality of small tubes is equalized or substantially equalized.

Each of the plurality of small tubes may extend vertically downward from the bottom surface of the reaction chamber and be combined with a large tube having a main discharge outlet at a downstream end thereof. The length of the tubule closest to the main discharge outlet is the longest. The length of the small tube furthest from the main discharge outlet is shortest. The smaller tubes closer to the main discharge outlet are longer than the smaller tubes further away from the main discharge outlet. This arrangement provides a simple way to provide natural drainage by gravity when suction is not sufficient and, at the same time, to provide suction equalization.

The large tube may be inclined such that the liquid collected by the small tube flows toward the main discharge outlet by gravity when no suction force is applied, and the inclination of the main discharge tube is determined by the vertical length of the longest small tube, the vertical length of the shortest small tube, and the interval between the longest small tube and the shortest small tube.

The inclination of the large tube, the length of the small tube connected to the large tube, the cross-sectional area of the small tube, and the distance of the small tube from the main discharge outlet may be adapted to obtain a uniform volumetric suction rate at the discharge orifice.

In another aspect of the present invention there is provided a cartridge test device suitable for use with a flow-through hybridization apparatus such as described herein, wherein the cartridge test device comprises a molded housing having a lattice formed thereon which forms a plurality of test cells or test compartments, and a porous permeant membrane mounted on the downstream side of the housing; wherein the lattice is adapted to facilitate separate hybridizations at a single test unit or test compartment.

In one embodiment, a sealing device is provided on the downstream side of the grid to provide a respective liquid seal to the individual cells when the cartridge test device is received within the pass-through detection device reaction chamber during use.

The seal on the downstream side of the grid may be adapted to prevent analyte flow from one test compartment to the other.

The portion of the permeable membrane in contact with the downstream side of the lattice may be subjected to a treatment, such as a heat treatment, to form a seal when the permeable membrane is attached to the downstream or bottom surface of the housing.

The permeant membrane is adapted to perform hybridization of multiple types of target analytes.

The cartridge test device with built-in permeable membrane provides a convenient solution for rapid testing while mitigating potential or cross contamination due to fewer personnel handling steps.

The permeable membrane may be attached to the bottom surface of the housing and when attached, sealing is performed.

The permeable membrane may comprise a hot melt material such as nylon and, when hot melt attached, the liquid seal is performed by hot melt.

The permeant membrane can be treated with one or more capture probes for capturing and displaying a specific or predetermined sequence of target analytes.

Examples of capture probes include any gene-specific nucleic acid oligonucleotide that is complementary to one or more target analyte sequences to form a single point for a single sequence or gene; for use in multiple spots in an array format, each spot having a specific sequence corresponding to a target sequence or gene in question, for detecting, for example, any single-type HPV (human papillomavirus) genotyping, i.e. type 16 or multi-genotyping assays, e.g. the 33-class GenoFlowArrayHPV assay already in use on the market, and additionally the P450-CYP2D6 genotyping reported in the patent pending application (PCT/IB2008/054044), and the HLA (human leukocyte antigen) genotyping classification (uspptono 7732138B 2). Other examples are, for example, simultaneous assays of multiple different classes of viral organisms, i.e., plate assays (pancelassay), which include HIV virus (human immunodeficiency virus), HBV virus (hepatitis b virus), and HCV virus (hepatitis c virus) in a transmembrane array or the like.

Drawings

Exemplary embodiments of the invention are described by way of example with reference to the accompanying drawings, in which:

FIG. 1 is an isometric view of a pass-through inspection apparatus according to the present invention;

FIGS. 2 and 3 are transverse isometric views of the pass-through detection apparatus of FIG. 1 with the housing removed;

FIG. 4 is an isometric view showing the reaction chamber of the pass-through detection apparatus of FIG. 3 with the cover of the chamber open;

FIG. 5 is an isometric view showing the reaction chamber of the pass-through detection apparatus of FIG. 3 mounted with the accommodated 4-module cartridge-type hybridization test device;

FIG. 5A is a plan view showing the relationship between the cartridge type testing device unit and the distribution of the discharge holes on the bottom surface of the reaction chamber of the apparatus shown in FIG. 1;

FIG. 6 shows the reaction chamber of FIG. 5, wherein the holding device is latched with the 4-module cartridge hybridization test device in the test position;

FIG. 7 shows the reaction chamber of FIG. 5 with the reclosable window closed to isolate the modular cartridge hybridization test device housed within the reaction chamber from the external environment;

FIG. 8 is a bottom isometric view of the reaction chamber of FIG. 4;

FIG. 9 is a partial exploded view of the reaction chamber of FIG. 4 with the integrated heat sink removed;

FIG. 10 is a side isometric view of the pass-through detection apparatus of FIG. 1 with the main housing removed and showing an air flow arrangement;

FIG. 11 is a longitudinal cross-sectional view of the apparatus of FIG. 10;

FIG. 12 is a top isometric view of the cast base of the reaction chamber of FIG. 1, showing the reaction chamber bottom surface;

FIG. 12A is a plan view showing an enlarged portion of the bottom surface of the reaction chamber;

FIG. 12B is a bottom isometric view of FIG. 11 showing a plurality of drain tubes connected to drain holes distributed on the bottom surface of the reaction chamber;

FIG. 13 is a schematic side view showing the reaction chamber connected to a drain net;

FIG. 14 is an enlarged schematic side view showing a portion of the drainage net of FIG. 13;

FIGS. 15 and 15A are first and second top isometric views showing a modular cassette hybridization test device suitable for use with the pass-through device of FIG. 1;

FIGS. 16 and 16A are first and second bottom isometric views of the modular cartridge hybridization test device of FIG. 15;

FIG. 17 is a longitudinal sectional view schematically showing that a modular cartridge-type hybridization testing device is mounted on a reaction chamber; and

fig. 18 is a schematic diagram illustrating the operation of the pass-through detection apparatus.

List of reference numerals

Detailed Description

The pass-through detection apparatus 100 shown in fig. 1 to 3 includes: a main housing 110 on which a reaction chamber 120 is mounted; a temperature regulation component 140; a power supply module; a data display screen 160; a control board 162; a vacuum suction pump 164; and other optional or peripheral devices. The peripheral devices include data communication interfaces such as a USB port 166, a silencer 163, and a vacuum reservoir 165.

The reaction chamber 120 is adapted to provide an enclosure of controlled environmental conditions to facilitate molecular reactions, such as between a target analyte and a reagent. As shown in detail in fig. 4, 5, 6, the reaction chamber 120 includes a case enclosure with a lid in which a metal holder 124 is installed to form a bottom surface. The shell enclosure includes: a main housing portion 122 molded from a hard plastic; a plastic cover or lid 126 hingedly connected to the main housing 122 and movable between an open position and a closed position about a hinge axis formed on one side of the main housing 122; and a latch frame 128, which is also hinged with the main housing 122 and shares the same hinge axis with the cover 126.

The reaction chamber 120 includes a main reaction compartment extending between the metal holder 124 and the lid 126 and enclosed by the side walls of the main housing portion 122. The lid 126 also includes a transparent window to allow a user to view the conditions within the main reaction compartment. The main reaction compartment is adapted to receive a modular cartridge hybridization test device (described in more detail below), and the hingedly attached lid 126 is adapted to facilitate insertion or loading of test objects into the reaction chamber for reaction, and removal or unloading of test objects upon opening the lid at the end of reaction. To facilitate reaction under controlled environmental conditions, and to mitigate potential contamination or harmful external or environmental effects, the lid will remain closed when a reaction occurs.

The base 130 of the reaction chamber is the upper surface of the metal holder 124. The metal bracket, which is an embodiment of a thermally conductive bracket, is made of stainless steel and is adapted to provide a thermal buffer such that the temperature at the base surface ("reaction surface") 130 can be precisely regulated and maintained within a target temperature range of ± 0.5 ℃ or even less during operation. A plurality of through holes are distributed throughout the metal base 130 to discharge the residual liquid by vacuum suction. The through-hole, which is an embodiment of a drain hole, is connected to a suction drain net to facilitate removal of excess or residual liquid within the reaction compartment. The drainage holes are distributed in a regular array or matrix to facilitate uniform or near uniform suction throughout the metal substrate when vacuum suction is applied during use. By providing and maintaining a uniform or substantially uniform negative pressure, in addition to draining residual liquid within the reaction chamber, the homogenized suction force also forms part of a controlled environment to control the flow of the tested target analyte through the permeable membrane of the cartridge testing device.

The portion of metal receptacle 124 below base surface 130 protrudes through a window formed in the bottom of main plastic housing enclosure 122 and has its bottom surface thermally coupled to temperature regulation assembly 140 (described below).

The temperature regulation component 140 is adapted to provide a temperature controlled reaction environment within the reaction compartment of the reaction chamber 120. This controlled environment is achieved by regulating the temperature of the metal holder, and thus, since the reaction surface is the upper surface of the metal holder 124, the temperature at the reaction surface 130 is also regulated.

Temperature regulation assembly 140 includes an electrical heating assembly and a forced air conditioning arrangement. The electric heating assembly comprises: a tab metal bracket 142 having a tab face 144 and a non-tab face 146; and a plurality of electrical heating elements 148, such as Peltier elements. Peltier elements 172 are mounted as an embodiment of an electrically driven thermal conditioning element or a thermoelectric element to provide electrical heating or cooling to the bottom surface of the metal holder 124 to provide the temperature controlled reaction surface 130. The bottom surface of metal bracket 124 is also thermally coupled to the non-finned surface of finned heat sink 142. A finned heat sink is used as an embodiment of the finned metal bracket, and the finned metal bracket is used as a heat exchanger to facilitate heat exchange with the outside air flow of the temperature conditioned air to facilitate temperature conditioning of the metal bracket 124.

To mitigate the undesirable thermal communication between metal bracket 124 and finned metal bracket 142, an insulator is placed around the space between metal bracket 124 and heat sink 142. In order to monitor the temperature distribution on the temperature-controlled reaction surface to maintain temperature uniformity on the reaction surface, a plurality of temperature sensors are distributed on the bottom surface of the metal holder to detect the temperature at different points on the metal base. The output signal of the temperature sensor is sent to an on-board microprocessor on the control board 162 for feedback control of the thermoelectric element 148 according to predetermined controlled conditions stored in the microprocessor.

While the distributed thermoelectric elements provide distributed heating or cooling to the reaction surface 130 to promote temperature uniformity across the reaction surface, the present apparatus further includes an additional forced air conditioning arrangement to provide thermal conditioning throughout the reaction surface by the flow of forced air to further improve temperature uniformity.

Referring to fig. 10 and 11, the forced air conditioning arrangement includes: an electric fan 180, such as an axial fan or a centrifugal fan, as an example of an air flow source adapted to generate a forced air flow; and an air duct 190 coupled to the air flow source for directing a forced air flow generated by the air flow source to temperature condition the reaction surface. The air duct comprises a first duct portion 191, a second duct portion 192, and a counter bore 193 between the first and second duct portions. The first pipe portion is elongated and extends from the air inlet 194 at the rear end of the pass-through detection device to the heat exchanger, the bottom surface of which also forms a portion of the first pipe portion and the end of the first pipe portion. The first duct portion is arranged to guide a forced air flow coming from the air inlet to form an air flow, and is arranged to be in contact with a bottom surface of the heat exchanger for heat exchange with the bottom surface of the heat exchanger immediately before reaching the counter hole. Ideally, the gas stream flows in a direction parallel or substantially parallel to the reaction surface at the tip when heat exchange occurs.

Thus, a slow fan can be used for more quiet operation while still providing sufficient heat exchange capacity, despite the slow airflow at the air inlet, the first duct portion is narrower closer to the end or counter bore end, so that the airflow is accelerated toward the heat exchange site to enhance the heat exchange capacity. To promote temperature uniformity throughout the entire base surface at the heat exchange, it should be noted that as the gas stream moves across the heat exchanger base surface, the heat content of the gas stream changes as a result of heat exchange therewith, the first conduit portion is tapered at its terminus, i.e., the end immediately adjacent the reversing orifice, so that as the gas stream approaches the terminus it accelerates further at the terminus to impart thermal uniformity across the entire heat exchanger. As shown more specifically in fig. 10 and 11, the end of the first pipe portion is constituted by the bottom surface of the heat exchanger and the baffle of the first pipe portion, which is thinner as it goes from the base side of the heat exchanger, which is the side closer to the intake port, to the end side, which is the closer to the counter bore. Since the bottom surface of the heat exchanger constitutes a substantially flat surface, the baffle end of the first conduit portion is gradually inclined upward toward the heat exchanger to form a wedge shape.

The reversing hole is located at the end of the first duct portion (i.e. the end of the first duct portion furthest from the air inlet) and immediately before the closed end of the air duct. The reversing holes provide an exit location for the exhaust air stream so that the exhaust air stream can exit the apparatus after passing over and contacting the bottom surface of the heat exchanger without encountering or interfering with the incoming air stream. The reverse hole is formed by a deflector formed by a concave closed end of the air duct and a terminal portion of the first duct portion, which is also a starting portion of the second duct portion. The curvature of the concave closed end is adapted to help direct the air flow so that it moves or passes more smoothly into the second conduit portion for venting.

The second conduit portion is located immediately below the first conduit portion 192 and is joined to the first conduit portion 191 by a reverse aperture 193. The second duct portion is parallel or substantially parallel to the first duct portion and constitutes a return path parallel or substantially parallel to the first duct portion, although the airflow is in the opposite direction to the incoming airflow. The second duct portion associated with the counter bore constitutes a return path so that the exhaust air flow flows in a backward direction without encountering or interfering with the incoming air flow, and, in view of the comfort of the user, does not have to exit at the front end of the device.

The air outlet 195 or the exhaust outlet is formed at the side of the second duct and is located at a position between the air inlet and the reverse hole so that the air flow can be discharged from the lateral side of the apparatus in a direction perpendicular to the incoming air flow, minimizing interference with the air inlet.

A plurality of through suction holes 129 are distributed in the base surface 124 for uniform suction across the entire base surface for sucking off residual liquid. These through-going holes are connected to the suction and discharge arrangement shown in detail in fig. 13 and 14. Each discharge arrangement includes a discharge net including a plurality of vertical branches or small discharge tubes 167 connected to a main discharge tube 168. The main exhaust pipe (or main pipe) is closed at one end, the open end thereof is connected to a suction source such as a vacuum pump, and a plurality of branched exhaust pipes are combined with the main pipe at a portion between the closed end and the open end. The large tube is inclined with respect to the reaction surface toward the open end so that residual liquid reaching the main drain tube will move toward the open end due to gravity even when the suction source is not operating. Therefore, the open end of the large tube forms the main discharge hole. Due to the inclination of the large tubes, and the generally horizontal arrangement of the reaction surfaces, the length of the small tubes is shortest at the closed end and longest at the open end. For tubules that are not located at the ends, the length of the tubules varies between the shortest and the longest, such that the length of the tubule closer to the open end is longer than the length of the tubule farther from the open end, or the tubule closer to the closed end is shorter than the tubule farther from the closed end.

In addition to draining residual liquid from the reaction chamber by gravity, the inclination of the large tubes, combined with the variable length of the small tubes, also facilitates providing uniform suction throughout the different through-holes associated with the same large tubes. In particular, by making the length of the small tube shorter the further away from the suction source end, the gradual decrease of the suction force at the end of the large tube is compensated for, in order to obtain a more uniform suction force and thus a more uniform suction rate, in particular a more uniform volumetric suction rate or suction speed.

To further enhance the uniformity of the pumping force throughout the reaction chamber, the reaction chamber is divided into a plurality of subsections such that each chamber subsection is under the pumping coverage of the discharge screen. The distributed drainage nets help to bring more uniform suction as the spatial coverage of each suction drainage net is smaller.

In another exemplary embodiment of the reaction chamber as shown in fig. 12, 12A and 12B, each of the suction holes 129 is enclosed by a net of discharge passages 132 respectively communicating with the suction holes 129. The network of discharge channels is arranged in a radial manner such that each channel extends radially from the suction aperture. By the radial or star-shaped arrangement of the discharge channels, it is facilitated that the suction forces originating from the suction apertures are distributed or spread across the surrounding channel footprint when the test object is in place. These channels are exposed slots and are suitably inclined towards the suction aperture to increase efficiency. A plurality of drain pipes 134 are also arranged below the metal brackets 124 to interconnect with a drain net.

Fig. 5A, and 15-17 illustrate an exemplary rapid cartridge test device suitable for use with the reaction chamber of the above-described flow-through assay apparatus for rapid and convenient analyte testing as described below.

The exemplary cartridge hybridization testing device 200 of FIG. 15 includes a main cartridge 210, a permeable membrane 220 attached to the main cartridge, and a sealing member 230. The main cartridge body is molded of plastic and includes a plurality of test compartment demarcations. The plastic body includes: main peripheral walls which collectively constitute an outer periphery of the case; and a partition wall which constitutes the respective test compartment in association with the peripheral wall. The peripheral walls and the partition walls form a lattice structure and together form a plurality of test wells 240, these test wells 240 forming test compartments after installation of the porous permeable membrane. The porous permeant is suitable for performing molecular analysis and can capture or retain capture probes for molecular identification, as is well known to those skilled in the art. As shown in fig. 5 and 5A, a single cartridge testing device may have 4 or 12 test wells or compartments. Of course, for most practical applications, other numbers of test compartments, e.g. 1 to 16, may be used.

To form a multi-compartment cartridge hybridization test device, the permeant film is affixed to the plastic body, for example, by melt bonding, such as hot melt bonding or ultrasonic bonding. Melt bonding is preferred because the melt bonding process also seals adjacent compartments so that analytes do not cross flow or leak into adjacent test compartments to avoid contamination of the analytes. Examples of suitable materials for the body and the permeable membrane are a nylon-based plastic body and a nylon-based permeable membrane, respectively, which have proven to have good fusing and sealing properties. To facilitate easy insertion and removal, a grip portion 250 is formed at a longitudinal end of the main cartridge body. The handle portions are integrally formed with the main body such that the lower ends of the handle portions are hingedly connected to provide resiliency to the handle portions. The resiliency of the handle portion allows the cartridge to rest tightly inside the reaction chamber and to be easily removed by pushing the free ends of the handle portion towards each other. In order to further fix the cartridge type testing device in the reaction chamber, a chamfered protrusion 252 is formed on the outer side of the handle portion as an example of the engaging means. The tapered projections are adapted to cooperate with latching means on the reaction chamber for latching engagement, thereby further securing the cartridge testing device against the reaction surface to achieve stable reaction conditions.

To provide further sealing to avoid cross-flow contamination between adjacent compartments and to provide an airtight seal on the bottom surface of the cartridge testing device to ensure uniform suction, an additional sealing ring in the form of a sealing template (e.g. a sealing template made of silicone rubber) is adhered to the bottom surface of the permeable membrane and is caused to travel along the compartments making up the lattice to form a sealing gasket.

As shown in FIGS. 5 and 5A, each cartridge is a modular cartridge type testing device, and a plurality of 4-cartridge type testing devices can be placed inside the reaction chamber. Because of the sealing gasket, which cooperates with the latching means of the reaction chamber, the reaction chamber can be operated with one to four modular cassette test devices. Although the modular cartridge testing devices shown here have the same physical dimensions, the modular cartridge testing device modules can obviously vary in size so that the reaction chamber can accommodate 1, 2 or 4 cartridge testing devices, or modular cartridge testing devices of different sizes. The modular cartridge tester design provides enhanced flexibility to enable testing using different types and numbers of cartridge testers independently.

As shown in fig. 5A, the cartridge testing device and the reaction chamber are arranged such that when the cartridge testing device is placed within the reaction chamber, a plurality of suction apertures are aligned with each test well or test compartment to promote uniform suction.

In use, the operating parameters of the pass-through apparatus are set, and a cartridge-type test device having a permeable membrane treated with a capture probe is placed in a reaction chamber and secured with a latch device. The target analyte is then applied to a cartridge testing device. The latching mechanism secures the cartridge testing device such that the permeable membrane is proximate to and pressed against the temperature controlled reaction surface to facilitate reaction in a controlled temperature environment when the reaction chamber is closed. When the device is set to operate under operating conditions, hybridization of the target analyte will occur under uniform suction to control the flow rate. Initial measurements showed that with the temperature regulation arrangement as described herein, a temperature uniformity of about ± 0.3% throughout the entire reaction surface was obtained.

When only a few samples are required to be tested, a single cartridge testing device or a selected plurality of cartridge testing devices is installed that is less than the maximum number of cartridge testing devices that can be housed within the reaction chamber. In this case, the cartridge hybridization test device is placed in the reaction chamber and latched into the operating position, together with one or a selected plurality of cartridge test devices, to divide the reaction chamber into a plurality of reaction compartments.

While the invention has been described with reference to the above embodiments, it will be understood by those skilled in the art that these embodiments are for illustrative purposes only and should not be used to limit the scope of the invention. For example, embodiments of the pass-through detection apparatus described herein include improvements relating to: temperature regulation is performed by means of an exemplary air duct arrangement with reversed apertures; temperature regulation is performed by means of a tapered air duct portion, distributed suction holes, a discharge net comprising inclined large tubes communicating with vertical small tubes (of varying length and arranged to obtain uniform suction), suction channels around the suction holes (to disperse suction to obtain suction uniformity); it is to be understood that such improvements or improved features may be applied independently or in combination without limitation.

Claims (14)

1. A cartridge test device adapted for use with a flow-through hybridization detection apparatus having a reaction chamber and a temperature-regulated reaction chamber bottom surface, characterized in that the cartridge test device comprises a molded main cartridge (210), a permeable membrane (220) affixed to a downstream side bottom surface of the main cartridge (210) and adapted to hybridize with passing analytes to capture and display a specific or predetermined sequence of a target analyte, and a sealing means (230) disposed downstream of the permeable membrane (220), wherein the main cartridge (210) forms at least one test compartment, the sealing means (230) adapted to cooperate with the reaction chamber bottom surface when used with the flow-through hybridization detection apparatus to provide a hermetic seal on the cartridge test device bottom surface to prevent analyte flow from the test compartment to an adjacent test compartment.

2. The cartridge testing device of claim 1, wherein said main cartridge body (210) comprises a grid constituting a plurality of test units or test compartments adapted to perform hybridization, said sealing means being arranged at a downstream side of said grid to provide a respective liquid seal to each test unit or test compartment to avoid cross-flow contamination between adjacent compartments or to prevent analyte flow from one test unit or test compartment to the other.

3. The cartridge type testing device according to claim 1 or 2, wherein said sealing means (230) is adhered to the bottom surface of said permeable membrane in the form of an additional sealing ring in the form of a sealing type plate and runs along the grid constituting the testing compartment to form a sealing gasket.

4. The cartridge-type test device according to any one of the preceding claims, wherein the permeable membrane approaches and presses against the bottom surface of the reaction chamber when used in combination with the pass-through hybridization detection apparatus.

5. The cartridge testing device of any one of the preceding claims, wherein the cartridge testing device has 1 to 16 test units or test compartments, including 4 or 12.

6. The cartridge testing device according to any one of the preceding claims, wherein the main cartridge (210) has a handle portion (250) at a longitudinal end thereof, the lower end of the handle portion (250) being hingedly connected to the main cartridge (210) and resilient.

7. The cartridge testing device of claim 6, wherein the resilience of the handle portion (250) causes the main cartridge body (210) to tightly rest inside the reaction chamber.

8. The cartridge testing device according to claim 6 or 7, wherein the handle portion (250) is integrally formed to the main cartridge body (210).

9. The cartridge type test device according to any one of the preceding claims, wherein the permeable membrane is a porous permeable membrane suitable for molecular analysis having one or more probes for capturing and displaying a specific or predetermined sequence of a target analyte.

10. The cartridge testing device according to any one of the preceding claims, wherein said main cartridge body (210) comprises a main peripheral wall constituting an outer periphery and partition walls cooperating with the main peripheral wall to constitute a plurality of test compartments, said main peripheral wall and said partition walls forming a lattice-like structure.

11. A flow-through assay device comprising a thermally regulated reaction chamber having a temperature regulated reaction chamber bottom surface and an air flow device and a cartridge test apparatus according to any preceding claim, wherein the reaction chamber comprises a temperature regulation surface adapted to facilitate flow-through hybridization under thermally regulated and controlled conditions and a thermal regulation component comprising an active electrothermal regulation device adapted to heat or cool the temperature regulation surface by conduction; wherein, on the bottom surface of the reaction chamber, a plurality of discharge holes suitable for discharging excessive or residual liquid in the reaction chamber are distributed; and wherein the reaction chamber and the cartridge testing device are arranged such that a test compartment constituted by a grid on the cartridge testing device is aligned with at least one suction aperture of the reaction chamber.

12. A flow-through apparatus according to claim 11, wherein the reaction chamber comprises a latching mechanism adapted to secure the cartridge testing device, the latching mechanism, in use, causing the permeable membrane to approach or press against a temperature-controlled bottom surface of the reaction chamber.

13. A flow-through apparatus according to claim 11 or 12, wherein the cartridge testing device and reaction chamber are arranged such that when the cartridge testing device is placed within the reaction chamber, a plurality of suction apertures are aligned with each test well or test compartment to promote uniform suction such that hybridization of the target analyte occurs under uniform suction.

14. A flow-through apparatus according to any one of claims 11 to 13, wherein the temperature of the bottom surface of the reaction chamber is maintained within a range of ± 0.5 ℃ or ± 0.3% or less of a target temperature during operation, and a plurality of discharge holes connected to a suction discharge net are distributed on the bottom surface of the reaction chamber so that an excess or residual liquid remaining on the bottom surface of the reaction chamber during use is discharged by vacuum suction.