NUCLEIC ACID SEPARATING APPARATUS
20200377845 ยท 2020-12-03
Assignee
Inventors
Cpc classification
B01L3/0275
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/0631
PERFORMING OPERATIONS; TRANSPORTING
B01F35/75465
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/16
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/025
PERFORMING OPERATIONS; TRANSPORTING
B01L3/0279
PERFORMING OPERATIONS; TRANSPORTING
G01N35/1011
PHYSICS
B01F31/22
PERFORMING OPERATIONS; TRANSPORTING
B01L2400/0487
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/023
PERFORMING OPERATIONS; TRANSPORTING
G01N2035/00277
PHYSICS
B01F35/71745
PERFORMING OPERATIONS; TRANSPORTING
C12N15/1017
CHEMISTRY; METALLURGY
G01N35/1009
PHYSICS
C12M47/06
CHEMISTRY; METALLURGY
B01F31/26
PERFORMING OPERATIONS; TRANSPORTING
International classification
G01N35/10
PHYSICS
Abstract
A nucleic acid separating apparatus includes: a stirring mechanism that stirs and resolving-processes a specimen and a reagent injected into a process tube; a pressurized gas supply mechanism that supplies a pressurized gas to a filter cartridge having a specimen liquid including nucleic acid, the specimen liquid being injected in the filter cartridge after the resolving-process, to separate and collect nucleic acid from the specimen liquid; a dispensing mechanism that injects the specimen, a reagent, or the specimen liquid after the resolving-process into the process tube and the filter cartridge; and a move mechanism that moves the pressurized gas supply mechanism and the dispensing mechanism, wherein the dispensing mechanism includes: a suction and discharge mechanism that is capable of attaching a dispensing chip to the suction and discharge mechanism and detaching the dispensing chip from the suction and discharge mechanism, the suction and discharge mechanism being capable of suctioning and discharging a liquid through the dispensing chip; and a liquid supply mechanism that discharges a liquid from a nozzle.
Claims
1. A nucleic acid separating apparatus comprising: a stirring mechanism that stirs and resolving-processes a specimen and a reagent injected into a process tube; a pressurized gas supply mechanism that supplies a pressurized gas to a filter cartridge having a specimen liquid including nucleic acid, the specimen liquid being injected in the filter cartridge after the resolving-process, to separate and collect nucleic acid from the specimen liquid; a dispensing mechanism that injects the specimen, a reagent, or the specimen liquid after the resolving-process into the process tube and the filter cartridge; and a move mechanism that moves the pressurized gas supply mechanism and the dispensing mechanism, wherein the dispensing mechanism comprises: a suction and discharge mechanism that is capable of attaching a dispensing chip to the suction and discharge mechanism and detaching the dispensing chip from the suction and discharge mechanism, the suction and discharge mechanism being capable of suctioning and discharging a liquid through the dispensing chip; and a liquid supply mechanism that discharges a liquid from a nozzle.
2. The nucleic acid separating apparatus according to claim 1, wherein the suction and discharge mechanism suctions or discharges a gas in the dispensing chip, and suctions or discharges a liquid into the dispensing chip, and wherein the suction and discharge mechanism comprises: a first dispensing chip attaching part that includes an O-ring having a first caliber at a tip of the first dispensing chip attaching part, the first dispensing chip attaching part being capable of attaching a first dispensing chip having the first caliber to the first dispensing chip attaching part and detaching the first dispensing chip from the first dispensing chip attaching part; and a second dispensing chip attaching part that includes an O-ring having a second caliber larger than the first caliber to be concentric with the O-ring having the first caliber, the second dispensing chip attaching part being capable of attaching a second dispensing chip having the second caliber to the second dispensing chip attaching part and detaching the second dispensing chip from the second dispensing chip attaching part.
Description
BRIEF DESCRIPTION OF DRAWINGS
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DETAILED DESCRIPTION
[0086] A nucleic acid separating apparatus according to a first aspect includes:
[0087] a stirring mechanism that stirs and resolving-processes a specimen and a reagent injected into a process tube;
[0088] a pressurized gas supply mechanism that supplies a pressurized gas to a filter cartridge having a specimen liquid including nucleic acid, the specimen liquid being injected in the filter cartridge after the resolving-process, to separate and collect nucleic acid from the specimen liquid
[0089] a dispensing mechanism that injects the specimen, a reagent, or the specimen liquid after the resolving-process into the process tube and the filter cartridge; and
[0090] a move mechanism that moves the pressurized gas supply mechanism and the dispensing mechanism,
[0091] wherein the dispensing mechanism includes: [0092] a suction and discharge mechanism that is capable of attaching a dispensing chip to the suction and discharge mechanism and detaching the dispensing chip from the suction and discharge mechanism, the suction and discharge mechanism being capable of suctioning and discharging a liquid through the dispensing chip; and [0093] a liquid supply mechanism that discharges a liquid from a nozzle.
[0094] Further, as a nucleic acid separating apparatus of a second aspect, in the first aspect, the suction and discharge mechanism suctions or discharges a gas in the dispensing chip, and suctions or discharges a liquid into the dispensing chip, and
[0095] wherein the suction and discharge mechanism includes: [0096] a first dispensing chip attaching part that includes an O-ring having a first caliber at a tip of the first dispensing chip attaching part, the first dispensing chip attaching part being capable of attaching a first dispensing chip having the first caliber to the first dispensing chip attaching part and detaching the first dispensing chip from the first dispensing chip attaching part; and; [0097] a second dispensing chip attaching part that includes an O-ring having a second caliber larger than the first caliber to be concentric with the O-ring having the first caliber, the second dispensing chip attaching part being capable of attaching a second dispensing chip having the second caliber to the second dispensing chip attaching part and detaching the second dispensing chip from the second dispensing chip attaching part.
[0098] A nucleic acid separating apparatus according to a third aspect includes:
[0099] a stirring mechanism that stirs and resolving-processes a specimen and a reagent injected into a process tube;
[0100] a pressurized gas supply mechanism that supplies a pressurized gas to a filter cartridge having a specimen liquid including nucleic acid, the specimen liquid being injected in the filter cartridge after the resolving-process, to separate and collect nucleic acid from the specimen liquid to a collection tube;
[0101] a dispensing mechanism that injects the specimen, a reagent, or the specimen liquid after the resolving-process into the process tube and the filter cartridge; and
[0102] a move mechanism that moves the pressurized gas supply mechanism and the dispensing mechanism,
[0103] a sample ID reading part that reads a sample ID of a sample container including the specimen;
[0104] a collection tube ID reading part that reads a collection tube ID of the collection tube; and
[0105] an ID managing part that manages the sample ID and the corresponding collection tube ID correlating these with each other.
[0106] Further, as a nucleic acid separating apparatus of a fourth aspect, in the third aspect, further may include a sample rack that stores the plural sample containers in row,
[0107] wherein a sample position ID may be provided at the position for each point to store the sample container in the each point of the sample rack, the sample position ID indicating the position for storing the sample container,
[0108] wherein a sample ID reading part may read the sample ID and the sample position ID from the direction that intersects the setting direction of the sample rack,
[0109] wherein the ID managing part may correlate the sample ID and the corresponding sample position ID each other.
[0110] Further, as a nucleic acid separating apparatus of a fifth aspect, in the third or fourth aspect, further may include a collection tube rack that stores the plural collection tubes in row,
[0111] wherein a collection tube position ID may be provided at the position for each point to store the collection tube in the each point of the collection tube rack, the collection tube position ID indicating the position for storing the collection tube,
[0112] wherein a collection tube ID reading part may read the collection tube ID and the collection tube position ID from the direction that intersects the setting direction of the collection tube rack,
[0113] wherein the ID managing part may correlate the collection tube ID and the corresponding collection tube position ID each other.
[0114] A nucleic acid separating apparatus according to each of embodiments will be described below with reference to the accompanying drawings. In the drawings, substantially same members are given the same reference numerals.
First Embodiment
[0115]
[0116] As depicted in
[0117] The arrangement in
[0118] This nucleic acid separating apparatus 10 includes the dispensing mechanism 21 and 22 that injects the specimen liquid and the reagent through the dispensing chips 32, and the pressured gas supply mechanism 23 that supplies a pressurized gas. Plural steps such as the dispensing and pressurizing can thereby be automated and not only the nucleic acid extraction step but also the traditionally manually performed pre-process steps can be automated.
[0119] The constituent members of the nucleic acid separating apparatus 10 will be described below.
[0120] <Dispensing Chip/Reagent Setting Part>
[0121]
[0122] Dispensing chips 32a, 32b, and 32c and 32d are respectively stored in dispensing chip racks 31a, 31b, and 31c. The dispensing chips 32a, 32b, 32c, and 32d may each have a capacity different from that of each other. For example, the dispensing chips 32a and 32c may each have the capacity of 1.2 ml and the dispensing chips 32b and 32d may each have the capacity of 10 ml. A dispensing chip ejecting part 33 is disposed as a point for detaching the unnecessary dispensing chip from the dispensing and pressurizing unit 15. The detached dispensing chip 32 is discarded into a dispensing chip discarding box 34. The dispensing chip discarding box 34 may be disposed in, for example, the lower portion of the apparatus.
[0123] Reagent containers 36a, 36b, 36c, 36d, and 36e and waste containers 37 for discarding the reagents are disposed on the reagent container tray 35. The reagents are, for example, a pre-process enzyme, a solution liquid, and ethanol that are used at the pre-process steps and a cleaning liquid and a collection liquid that are used at the nucleic acid extracting step. For example, the pre-process enzyme may be put in the reagent container 36a, the solution liquid may be put in the reagent container 36b, ethanol may be put in the reagent container 36c, and the collection liquid may be put in the reagent container 36d. Which reagent is put in which reagent container only has to be properly set taking into consideration the convenience. A reagent bottle 36f may separately be disposed for the cleaning liquid in the lower portion of the apparatus as depicted in
[0124] <Sample Setting Part>
[0125]
[0126] Sample rack presence or absence sensing sensors 43a, 43b, and 43c of the sample racks 41a, 41b, and 41c are respectively disposed at the back ends in a setting direction 44 of the sample racks 41a, 41b, and 41c. The sample rack presence or absence sensing sensors 43a, 43b, and 43c respectively detect whether the sample racks 41a, 41b, and 41c are set. As to each of the sample racks 41a, 41b, and 41c, for each corresponding lane, a shape engaging with each other may be disposed in the back end portions. Each of the sample racks 41a, 41b, and 41c thereby does not engage with the non-corresponding lane at its end portion and any mistake of inserting into a wrong lane can be suppressed.
[0127] As depicted in
[0128] <Specimen>
[0129] In, for example, the diagnosis field, each of such items collected as test substances is handled as a specimen, as biological fluids such as the whole blood, blood plasma, blood serum, urine, feces, semen, and saliva, or a plant (or a portion thereof), an animal (or a portion thereof), or a melt of each thereof, and a solution prepared from a biological material such as a homogenate.
[0130] <Warming/Stirring Part>
[0131]
[0132] The warming/stirring part 13 can stir and warm the liquid in each of the process tubes 53. A process tube rack 52 storing the process tubes 53 in the stirring unit 51 is disposed in the warming/stirring part 13. In the example in
[0133] As depicted in
[0134] As depicted in
[0135] As depicted in
[0136] <Nucleic Acid Extracting Part>
[0137]
[0138] The nucleic acid extracting part 14 includes filter cartridge racks 61a, 61b, and 61c each storing therein the filter cartridges 63, and collection tube racks 62a, 62b, and 62c each storing therein the collection tubes 66. The filter cartridge racks 61a, 61b, and 61c each store therein eight filter cartridges 63. The collection tube racks 62a, 62b, and 62c each store therein eight collection tubes 66. In the example in
[0139] As depicted in
[0140] The filter cartridge racks 61a, 61b, and 61c may each have the shape engaging with each other disposed at the back end portion for each corresponding lane. The filter cartridge racks 61a, 61b, and 61c each thereby do not engage in its end portion in any non-corresponding lane and any mistake of inserting into a wrong lane can therefore be suppressed. Similarly, the collection tube racks 62a, 62b, and 62c may each have the shape engaging with each other disposed at its back end portion for each corresponding lane. The collection tube racks 62a, 62b, and 62c each thereby do not engage in its end portion in any non-corresponding lane and any mistake of inserting into a wrong lane can therefore be suppressed.
[0141] As depicted in
[0142] <Filter>
[0143] As depicted in
[0144] Examples of the porous film of an organic material including a hydroxyl group include a surface-saponified substance of acetylcellulose described in Japanese Laid-Open Patent Publication No. 2003-128691. This acetylcellulose may be any one of monoacetylcellulose, diacetylcellulose, and triacetylcellulose while triacetylcellulose is especially advantageously used. In this case, the amount (the density) of hydroxyl groups on the solid phase surface can be controlled using the degree of the saponification process (saponification degree). To improve the separation efficiency of nucleic acid, it is preferred that the amount (the density) of hydroxyl groups be large. For example, in the case of acetylcellulose such as triacetylcellulose, the saponification ratio is preferably 5% or higher and further preferably 10% or higher. To increase the surface area, a saponification process is applied to the porous film of acetylcellulose. The spatial amount (the density) of hydroxyl groups can be controlled by a combination of the degree of the saponification process (the saponification degree) and the pore diameter of the porous film. In this case, the porous film may be a porous film that is symmetrical for its front side and its back side while a porous film that is not symmetrical for its front side and its back side can advantageously be used.
[0145] In addition, examples of the porous film of the organic material including a hydroxyl group include porous films each formed by polyhydroxyethyl acrylic acid, polyhydroxyethyl methacrylic acid, polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, polymethacrylic acid, polyoxyethylene, acetylcellulose, a mixture of acetylcelluloses each having an acetyl value different from that of each other, or the like while a porous film of an organic material that has a polysaccharide structure can advantageously be used. Especially, the porous film of an organic polymer including a mixture of acetylcelluloses each having an acetyl value different from that of each other can advantageously be used, and such mixtures can advantageously be used as a mixture of triacetylcellulose and diacetylcellulose, a mixture of triacetylcellulose and monoacetylcellulose, a mixture of triacetylcellulose, diacetylcellulose, and monoacetylcellulose, and a mixture of diacetylcellulose and monoacetylcellulose.
[0146] Examples of the filter 64 include a porous film including an organic material formed by applying a saponification process to a mixture of acetylcelluloses each having an acetyl value different from that of each other and, for example, an oxide of each of the above mixtures of acetylcelluloses can advantageously be used. The saponification process is conducted by bringing acetylcellulose into contact with a saponification process liquid (such as, for example, sodium hydroxide), and a portion of acetylcellulose in contact with the saponification process liquid becomes regenerated cellulose and hydroxyl groups are introduced thereinto. To improve the separation efficiency of nucleic acid, it is more advantageous to introduce a larger number of hydroxyl groups. For example, the saponification ratio is preferably about 5% or higher and more preferably about 10% or higher.
[0147] A porous film of regenerated cellulose can advantageously be used as the filter 64. The regenerated cellulose is farmed by converting the surface or the whole of solid acetylcellulose into cellulose by applying a saponification process thereto, and is different from the original cellulose in the points of the crystal condition and the like.
[0148] Other examples of the filter 64 include a porous film including a silica compound as a porous film of an inorganic material including a hydrophilic group such as, for example, a glass filter. Other examples thereof include a porous silica thin film as described in Japanese Patent Publication No. 3058342. This porous silica thin film can be produced by spreading out on a substrate a developing solvent of a cation-type amphiphilic substance having a bimolecular membrane forming function, thereafter removing the solvent from the liquid film on the substrate, thereby preparing a multi-layer bimolecular membrane thin film of the amphiphilic substance, bringing this film into contact with a solution including a silica compound, and removing by extracting the multi-layer bimolecular membrane thin film.
[0149] A porous film is usable as the filter 64, whose thickness is 10 m to 500 m and preferably 50 m to 250 m. A porous film is usable whose minimal pore diameter is 0.22 m or larger and more preferably 0.5 m or larger. A porous film is usable whose ratio of its maximal pore diameter to its minimal pore diameter is 2 or greater and further preferably 5 or greater. A porous film is usable whose void ratio is 50% to 95% and further preferably 65% to 80%.
[0150] The one filter 64 may be used while the plural filters 64 may also be used. The plural filters 64 may be same ones or may be different ones. The filters 64 may be a combination of an inorganic material filter(s) and an organic material filter(s). Examples thereof include, for example, a combination of a glass filter and a regenerated cellulose porous film. The plural filters may be a combination of an inorganic material filter(s) and an organic material porous film(s) having no adsorptive property for nucleic acid. Examples thereof include, for example, a combination of a glass filter(s) and a porous film(s) of nylon or polysulfone.
[0151] <Collection Tube>
[0152] As depicted in
[0153] <About Moving of Filter Cartridge>
[0154] As depicted in
[0155] For example, the filter cartridges 63 are moved in the following three stages.
[0156] 1) The filter cartridges 63 are first moved to be lifted by the filter cartridge racks 61a, 61b, and 61c.
[0157] 2) The filter cartridges 63 are next horizontally moved from the position above the filter cartridge racks 61a, 61b, and 61c to a position above the collection tube racks 62a, 62b, and 62c.
[0158] 3) The filter cartridges 63 are next moved to be descended from the position above the collection tube racks 62a, 62b, and 62c.
[0159] The filter cartridges 63 can be moved from the filter cartridge racks 61a, 61b, and 61c to the collection tube racks 62a, 62b, and 62c by the above operations. The filter cartridges 63 may be fixed and the collection tubes 66 and the waist tubes 65 may be moved.
[0160] <Dispensing and Pressurizing Unit>
[0161]
[0162] The dispensing and pressurizing unit 15 includes dispensing mechanisms 21a, 21b, 22a, and 22b that each supply a liquid for containers whose upper face is open in the vertical direction (the process tubes 53 and the filter cartridges 63), pressurized gas supply mechanisms 23a and 23b that each supply a pressurized gas into the containers 63, and a move mechanism 24 that moves the overall body to a position above the containers 53 and 63 in the vertical direction. The dispensing mechanism includes a suction and discharge mechanism 22a and 22b, and liquid supply mechanisms 21a and 21b that each directly discharge a liquid from a nozzle. The suction and discharge mechanism 22a and 22b is capable of attaching thereto and detaching therefrom dispensing chips 32a and 32b. The suction and discharge mechanism 22a and 22b discharges a liquid such as a specimen liquid or a reagent through the dispensing chips 32a and 32b to the containers 53 and 63, and suction a liquid from the containers 53 to the dispensing chips 32a and 32b. The liquid supply mechanisms 21a and 21b can each supply a liquid to the containers 53 and 63.
[0163] The dispensing and pressurizing unit 15 may have a work sensing sensor 28 disposed therein. The work sensing sensor 28 senses whether the sample containers 42, the dispensing chips 32, the reagent containers 36, the process tubes 53, the filter cartridges 63, the collection tubes 66, and the like are each set at the predetermined positions for the predetermined quantity. A photoelectric sensor formed by combining a CMOS sensor and a laser light source, or the like is usable as the work sensing sensor 28. The work sensing sensor 28 can be moved by the move mechanism 24 to the vicinity of a position above each of the works to be sensed and can sense presence or absence of the work. It is preferred that the work sensing sensor check each of the works before the start of the pre-process steps.
[0164] The dispensing and pressurizing unit 15 includes two sets of dispensing mechanisms 21a, 21b, 22a, and 22b, and that of pressurized gas supply mechanism 23a and 23b while the quantity is not limited to two sets and one set may be included. Otherwise, three or more sets may be included. The dispensing mechanisms 21a and 22a and the pressurized gas supply mechanism 23a, and the dispensing mechanisms 21b and 22b, and the pressurized gas supply mechanism 23b may be adapted to each be movable in a z-axis direction separately from each other. Each individual dispensing operation is enabled by the fact that the dispensing mechanisms and the like are each independently movable in the z-axis direction as above. For example, when the two sets of dispensing mechanisms are present and an odd number of sample containers 42 to be processed are present, the dispensing can first be executed for two sets thereof at one time, and the dispensing from the finally remaining one sample container can be executed by operating only one dispensing mechanism. When the dispensing mechanisms are each not independent, complicated processing is necessary for attachment of the dispensing chips, and the like because all the dispensing mechanisms similarly move.
[0165] As depicted in
[0166] The members constituting the dispensing and pressurizing unit 15 may each be disposed separately from each other without constituting the one unit.
[0167] <Suction and Discharge Mechanism>
[0168] The suction and discharge mechanism (a pipette) 22a and 22b is characterized in that the mechanism does not dispense the cleaning liquid and the like each through a pipe directly connected to a bottle including the cleaning liquid or the like but each inject the liquid through the dispensing chip 32 attached to the tip thereof. A problem is traditionally present that many dispensing mechanisms need to be prepared each for a type of liquid such as the cleaning liquid or the collection liquid. A method is present according to which one discharge nozzle is set at the tip and the liquid to be discharged is switched by a switching valve or the like. In this case, however, to avoid any mixing of the liquids during the switching, cleaning using the liquid to be used after the switching, or the like needs to be executed and discarding of the wasted liquid occurs. In contrast, according to the suction and discharge mechanism 22a and 22b, dispensing can be executed changing the dispensing chip 32 for each of the liquids to be dispensed. The substantially one set of suction and discharge mechanism 22a and 22b can dispense various liquids. Any gas in the dispensing chip 32 can be suctioned or discharged, and a liquid can be suctioned and discharged in the dispensing chip 32.
[0169] As depicted in
[0170] <Liquid Supply Mechanism>
[0171]
[0172] <Pressurizing Head (Pressurized Gas Supply Mechanism)>
[0173]
[0174] The pressurizing heads 23a and 23b are respectively connected to air filters 81a and 81b that are disposed at a ventilating opening through pulse motors 82a and 82b, pressurizing pumps 83a and 83b, and pressure sensors 86a and 86b. The set range of the pressure is, for example, 30 kPa to 160 kPa. For example, air or nitrogen is usable as the pressurized gas.
[0175] The pressurizing heads 23a and 23b are arranged to avoid their interference on the operations of the suction and discharge mechanism 22a and 22b and the liquid supply nozzles 21a and 21b.
[0176] <Move Mechanism>
[0177] The move mechanism 24 can move the overall dispensing and pressurizing unit 15 to the position above the containers 53 and 63 in the vertical direction. For example, as depicted in
[0178] <Control Unit>
[0179] The control unit 16 controls the dispensing chip/reagent setting part 11, the sample setting part 12, the warming/stirring part 13, the nucleic acid extracting part 14, and the dispensing and pressurizing unit 15 that constitute the nucleic acid separating apparatus 10. The control part 16 may be, for example, a computer. The computer only has to include the physical components such as, for example, a CPU 1, a memory 2, a storing part 3, a displaying part 4, an input part 5, and an interface 6.
[0180] <Nucleic Acid Separation Method>
[0181]
[0187] The resolving process is processing executed using a water solution including a reagent (such as a solution including, for example, chaotropic salt or guanidine salt, a surfactant, and a protein-degrading enzyme) that resolves the cell membrane and the nucleic membrane and thereby causes the nucleic acid to be soluble and, for example, when the specimen to be processed is the whole blood, the red blood cells and various types of protein are degraded to be low molecules to prevent any non-specific adsorption to and any clogging of the filter 64, and the white blood cells and the nucleic membranes are resolved to cause the nucleic acid to be extracted, to be soluble. Examples of a water-soluble organic solvent include ethanol, isopropanol, propanol, or the like and, among these, ethanol is advantageously used. The concentration of the water-soluble organic solvent is preferably 5% by weight to 90% by weight and further preferably 20% by weight to 60% by weight. The addition concentration of ethanol is especially advantageously set to be as high as possible to the extent that no aggregation is formed.
[0188] The whole blood, or a test substance including cells or viruses is resolving-processed and this produces the state where the water-soluble organic solvent is added to a solution that has nucleic acid dispersed in the liquid. [0189] (6) The reagent (ethanol) is suctioned from the reagent container 36 to the dispensing chip 32 (
[0194] <Cleaning Liquid>
[0195] The cleaning liquid has a function of washing away any impurity in the specimen liquid, adhering to the filter 64 together with nucleic acid, and has a composition for any impurity to depart leaving the adsorption of nucleic acid as it is. The cleaning liquid includes, for example, a base agent and a buffer agent, and a water solution when necessary that includes a surfactant. The main agent is methanol, ethanol, isopropanol, n-isopropanol, butanol, acetone, or the like. In the water solution, the main agent is about 10% by weight to 100% by weight, preferably 20% by weight to 100% by weight, and further preferably 40% by weight to 80% by weight in the water solution. [0196] (11) The filter cartridge 63 is filtered under pressure (
[0203] <Collection Liquid>
[0204] The collection liquid advantageously has a low salt concentration. Especially, a solution whose salt concentration is 0.5 M or lower such as, for example, purified distilled water or a TE buffer is used. pH of the collection liquid is preferably pH2 to pH11 and further preferably pH5 to pH9. [0205] (18) The filter cartridge 63 is filtered under pressure (
[0206] The collection liquid including nucleic acid can be collected by the above operation. For nucleic acid in the collection tube 66, a nucleic acid analysis process and the like are next executed.
[0207] According to the nucleic acid separating apparatus 10 and the nucleic acid separation method, for the plural sample containers 42, steps from the pre-process steps to the nucleic acid extraction step are sequentially and continuously executed for each of the sample containers. The manually executed traditional pre-process steps therefore do not need to be executed.
[0208] In this embodiment, plural sets of the sample containers 42, the process tubes 53, and the filter cartridges 63 are included while the number of sets is not limited to these. For example, the nucleic acid separating apparatus 10 according to the first embodiment is applicable to one set of the sample container 42, the process tube 53, and the filter cartridge 63.
[0209] The nucleic acid separating apparatus 10 may include a UV applying part (not depicted) that disinfects by applying a UV light beam to the inside of the apparatus. The UV applying part may be configured to apply a UV light beam moving inside the apparatus in, for example, the x- and the y-directions. The inside of the apparatus can thereby be maintained to be clean.
Second Embodiment
[0210] The planar configuration of a nucleic acid separating apparatus 10 according to the second embodiment seen through a top board thereof, and the configuration of the nucleic acid separating apparatus 10 seen through a front panel thereof are substantially similar to those of the first embodiment. For example, the configurations are as depicted in
[0211] The nucleic acid separating apparatus 10 includes the dispensing chip/reagent setting part 11, the sample setting part 12, the warming/stirring part 13, the nucleic acid extracting part 14, the dispensing and pressurizing unit 15, the control part 16, and an ID managing part 17. The ID managing part 17 includes a sample ID reading part 18 and a collection tube ID reading part 19. The sample ID reading part 18 reads a sample ID of a sample container 42. The collection tube ID reading part 19 reads a collection tube ID of a collection tube. The ID managing part 17 manages the sample ID and the corresponding collection tube ID correlating these with each other.
[0212] According to the nucleic acid separating apparatus 10, the ID managing part 17 can manage the sample ID of the sample container 42 and the collection tube ID of the corresponding collection tube 66 correlating these with each other. Any mistaken recognition for the sample container 42 and the corresponding collection tube 66 can thereby be suppressed.
[0213] As depicted in
[0214] The arrangement in
[0215] The nucleic acid separating apparatus 10 includes the dispensing mechanism 21 and 22 that injects the specimen liquid and the reagent through the dispensing chip 32, and the pressurized gas supply mechanism 23 that supplies the pressurized gas. The plural steps such as the dispensing and the pressurizing can thereby be automated, and not only the nucleic acid extraction step but also the traditionally manually executed pre-process steps can be automated.
[0216] The constituent members of the nucleic acid separating apparatus 10 will be described below. The configurations substantially same as those of the nucleic acid separating apparatus according to the first embodiment will properly not be again described.
[0217] <Dispensing Chip/Reagent Setting Part>
[0218] The overall configuration of the dispensing chip/reagent setting part 11 of the nucleic acid separating apparatus 10 is similar to that in the plan diagram of
[0219] The dispensing chips 32a, 32b, and 32c and 32d are respectively stored in the dispensing chip racks 31a, 31b, and 31c. The dispensing chips 32a, 32b, 32c, and 32d, the reagent containers 36a, 36b, 36c, 36d, and 36e, the waste container 37 for discarding the reagents, and the reagents are substantially similar to those of the first embodiment and will not again be described.
[0220] <Sample Setting Part>
[0221] The configuration of the sample setting part 12 of the nucleic acid separating apparatus 10 is similar to that in the planar diagram of
[0222] The sample racks 41a, 41b, and 41c, and the sample containers 42 are substantially similar to those of the first embodiment and will not again be described.
[0223] <Specimen>
[0224] For example, in the diagnosis field, each of such items collected as test substances is handled as a specimen, as biological fluids such as the whole blood, blood plasma, blood serum, urine, feces, semen, and saliva, or a plant (or a portion thereof), an animal (or a portion thereof), or a melt of each thereof, and a solution prepared from a biological material such as a homogenate.
[0225] <Sample ID Reading Part>
[0226]
[0227] As depicted in
[0228] As depicted in
[0229] As depicted in
[0230] The read sample ID and the read sample position ID are correlated with the corresponding collection tube ID by the ID managing part 17. Based on this correlation, the specimen accommodated in the sample container 42 is resolving-processed and the collection liquid including nucleic acid after the processing is accommodated in the corresponding collection tube 66.
[0231] <Warming/Stirring Part>
[0232] The configuration of the warming/stirring part 13 of the nucleic acid separating apparatus 10 is similar to that in the plan diagram of
[0233] The warming/stirring part 13, the process tube 53, and the stirring and warming thereof are substantially similar to those of the first embodiment and will not again be described.
[0234] <Nucleic Acid Extracting Part>
[0235] The configuration of the nucleic acid extracting part 14 of the nucleic acid separating apparatus 10 is similar to that in the plan diagram of
[0236] The nucleic acid extracting part 14, the filter cartridge 63, and the collection tube 66 are substantially similar to those of the first embodiment and will not again be described.
[0237] <Filter>
[0238] As depicted in
[0239] <Collection Tube>
[0240] As depicted in
[0241] <Collection Tube ID Reading Part>
[0242]
[0243] As depicted in
[0244] As depicted in the bottom diagram of the collection tube rack 62a of
[0245] As depicted in
[0246] <Modification Example>
[0247]
[0248] As depicted in
[0249] The read collection tube ID is correlated with the corresponding sample ID by the ID managing part 17. Based on this correlation, the collection liquid including nucleic acid originated from the specimen accommodated in the sample container 42 of the corresponding sample ID is accommodated in the corresponding collection tube 66.
[0250] When the collection tube position ID indicating the position of each of the collection tubes 66 is already read, the ID managing part 17 can correlate the collection tube ID and the corresponding collection tube position ID with each other. The position of each of the collection tubes 66 can thereby be identified in the collection tube racks 62a, 62b, and 62c.
[0251] <About Move of Filter Cartridge>
[0252] The state where the filter cartridge 63 is moved to the position above the collection tube 66 for collection of nucleic acid, to separate nucleic acid in the nucleic acid extracting part 14 is similar to that in
[0253] The filter cartridges 63 need to be moved to the position above the collection tubes 66 before the collection of nucleic acid. The move of the filter cartridges 63 is substantially similar to that of the first embodiment and will not again be described.
[0254] <Dispensing and Pressurizing Unit>
[0255] The configuration of the dispensing and pressurizing unit 15 of the nucleic acid separating apparatus 10 is similar to that in the schematic perspective diagram of
[0256] The dispensing and pressurizing unit 15 and the receiving tray are substantially similar to those of the first embodiment and will no again be described.
[0257] <Suction and Discharge Mechanism>
[0258] The suction and discharge mechanism (the pipette) 22a and 22b is characterized in that the mechanism does not dispense the cleaning liquid and the like each through a pipe directly connected to a bottle including the cleaning liquid or the like but injects the liquid through the dispensing chip 32 attached to the tip thereof The suction and discharge mechanism (the pipette) 22a and 22b is substantially similar to that of the first embodiment and will not again be described.
[0259] <Liquid Supply Mechanism>
[0260]
[0261] <Pressurizing Head (Pressurized Gas Supply Mechanism)>
[0262]
[0263] The pressurizing heads 23a and 23b are respectively connected to the air filters 81a and 81b that are disposed at the ventilating opening through the pulse motors 82a and 82b, the pressurizing pumps 83a and 83b, and the pressure sensors 86a and 86b. The set range of the pressure is, for example, 30 kPa to 160 kPa. For example, air or nitrogen is usable as the pressurized gas.
[0264] The pressurizing heads 23a and 23b are arranged to avoid their interference on the operations of the suction and discharge mechanism 22a and 22b, and the liquid supply nozzles 21a and 21b.
[0265] <Move Mechanism>
[0266] The move mechanism 24 can move the overall dispensing and pressurizing unit 15 to the position above the containers 53 and 63 in the vertical direction. For example, as depicted in
[0267] <Control Unit>
[0268] The control unit 16 controls the dispensing chip/reagent setting part 11, the sample setting part 12, the warming/stirring part 13, the nucleic acid extracting part 14, and the dispensing and pressurizing unit 15 that constitute the nucleic acid separating apparatus 10. The control part 16 may be, for example, a computer. The computer only has to include the physical components such as, for example, the CPU 1, the memory 2, the storing part 3, the displaying part 4, the input part 5, and the interface 6. The ID managing part 17 may be realized by hardware and software that are commonly used with the control part 16.
[0269] <Nucleic Acid Separation Method>
[0270]
[0278] The resolving process is processing executed using water solution including a reagent (such as, for example, a solution including chaotropic salt or guanidine salt, a surfactant, and a protein-degrading enzyme) that melts the cell membrane and the nucleic membrane and thereby causes the nucleic acid to be soluble. This resolving process is substantially similar to that of the first embodiment and will no again be described. [0279] (8) The reagent (ethanol) is suctioned from the reagent container 36 to the dispensing chip 32 (
[0284] <Cleaning Liquid>
[0285] The cleaning liquid has a function of washing away any impurity in the specimen liquid adhering to the filter 64 together with nucleic acid, and has a composition for any impurity to depart leaving the adsorption of nucleic acid as it is. The cleaning liquid is similar to that of the first embodiment and will not again be described. [0286] (13) The filter cartridge 63 is filtered under pressure (
[0293] <Collection Liquid>
[0294] The collection liquid advantageously has a low salt concentration. Especially, a solution having a salt concentration of 0.5 M or lower such as, for example, purified distilled water or a TE buffer is used. pH of the collection liquid is preferably pH2 to pH11 and further preferably pH5 to pH9. [0295] (20) The filter cartridge 63 is filtered under pressure (
[0296] The collection liquid including nucleic acid can be collected by the above operation. For nucleic acid in the collection tube 66, a nucleic acid analysis process and the like are next executed.
[0297] According to the nucleic acid separating apparatus 10 and the nucleic acid separation method, for the plural sample containers 42, steps from the pre-process steps to the nucleic acid extraction step are sequentially and continuously executed for each of the sample containers. The manually executed traditional pre-process steps therefore do not need to be executed.
[0298] In this second embodiment, plural sets of the sample container 42, the process tube 53, and the filter cartridge 63 are included while the number of sets is not limited to these. For example, the nucleic acid separating apparatus 10 according to the second embodiment is applicable to one set of the sample container 42, the process tube 53, and the filter cartridge 63.
[0299] The nucleic acid separating apparatus 10 may include a UV applying part (not depicted) that disinfects by applying a UV light beam to the inside of the apparatus. The UV applying part may be configured to apply a UV light beam moving inside the apparatus in, for example, the x- and the y-directions. The inside of the apparatus can thereby be maintained to be clean.
[0300] This disclosure includes a proper combination of any optional embodiment(s) and/or Example(s) of the above various embodiments and/or Examples, and the effects to be achieved by the embodiment(s) and/or Examples can be achieved.
[0301] According to the nucleic acid separating apparatus of the present invention, a dispensing mechanism that dispenses a specimen, a reagent, or a specimen liquid after a resolving process through a dispensing chip, and a pressurized gas supply mechanism that supplies a pressurized gas are included therein. Plural steps such as the dispensing and the pressurizing can thereby be automated, and not only a nucleic acid extraction step but also the traditionally manually executed pre-process steps can be automated.
[0302] According to the nucleic acid separating apparatus of another example of the present invention, an ID managing part can correlate a sample ID of a sample container, and a collection tube ID of the corresponding collection tube with each other. Any mistaken recognition for the sample container and the corresponding collection tube can thereby be suppressed.
REFERENCE NUMERALS LIST
[0303] 10 nucleic acid separating apparatus
11 dispensing chip/reagent setting part
12 sample setting part
13 warming/stirring part
14 nucleic acid extracting part
15 dispensing and pressurizing unit
16 control part
17 ID managing part
18 sample ID reading part
19 collection tube ID reading part
21, 21a, 21b reagent addition nozzle (liquid supply mechanism)
22, 22a, 22b suction/discharge nozzle (suction and discharge mechanism)
23, 23a, 23b pressurizing head (pressurized gas supply mechanism)
24, 24a, 24b, 24c move mechanism
25 receiving tray
26a, 26b first dispensing chip attaching part
27a, 27b second dispensing chip attaching part
28 work sensing sensor
31a, 31b, 31c dispensing chip rack
32, 32a, 32b, 32c, 32d dispensing chip
33 dispensing chip ejector part
34 dispensing chip discarding box
35 reagent container tray
36, 36a, 36b, 36c, 36d, 36e, 36f reagent container
41a, 41b, 41c sample rack
42, 42a, 42b, 42c sample container (blood collection tube)
43a, 43b, 43c sample rack presence or absence sensing sensor
44 sample rack setting direction
45a bar code (sample ID)
45b bar code (sample position ID)
46 code reader
47 red LED light beam
51 stirring mechanism (stirring unit)
52 process tube rack
53 process tube (lysate tube)
54 neck portion
55 bottom portion
56 rotation direction
58 heater
61a, 61b, 61c filter cartridge rack
62a, 62b, 62c collection tube rack
63 filter cartridge
64 filter
65 waste tube
66 collection tube
67 move to be lifted
58 move in the horizontal direction
69 downward move
71 tube pump
72 pipe
74 ID reading dedicated lane
75 two-dimensional code
76 code reader
77 code reader move lane
81a, 81b air filter
82a, 82b pulse motor
83a, 83b pressurizing pump
86a, 86b pressure sensor