JP2009022241A - Biochemical reactor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a biochemical reactor capable of preventing the contamination of a testing sample held in the reactor, without having to use a complex mechanism. <P>SOLUTION: The biochemical reactor is provided with at least one transporting part to transport a substance into the reactor, at least one cell formed by covering the inside of the reactor with a shielding plate, at least one transport port formed on the cell, a shielding member for shielding the transport port, and an atmosphere purging means formed on the cell for purging the atmosphere in the cell. The shielding member is placed at the transport part, and the atmosphere purging means purges the atmosphere in the cell, when the cell is in a shielded state. <P>COPYRIGHT: (C)2009,JPO&amp;INPIT

Description

  The present invention relates to a biochemical reaction apparatus capable of preventing contamination.

  In tailor-made medicine, genetic testing is one of the means to back up the diagnosis. The reliability of genetic testing is very important in order to produce test results that lead to accurate diagnosis.

  As a biochemical reaction device applied to genetic testing, for example, there is a DNA testing device using a DNA chip. A DNA chip is a substrate in which many types of DNA fragments are pasted on a substrate such as glass as probe DNA, and many types of DNA can be examined at once. The process includes, for example, an extraction step for extracting nucleic acid from a specimen, an amplification step for amplifying nucleic acid, a hybridization step for performing a hybridization reaction between a nucleic acid and a probe, a detection step for detecting a hybridization reaction, and a step for determining a result. It is done in order.

  As an example, in the case of examining blood DNA, each step of extraction, amplification, hybridization, detection, and result determination will be described in detail.

  In the extraction process, DNA is extracted, separated and purified from a blood sample. Pipette blood into the extraction solution and incubate at about 50 ° C. for several minutes. As a result of incubation, DNA is extracted from the cells. Magnetic beads and column filters are generally used as means for separating and purifying DNA from the extracted DNA extraction solution. For purification with magnetic beads suitable for automation, a DNA extraction solution is dispensed into a magnetic bead solution, the solution is stirred with a pipette for several minutes, and is allowed to stand gently for a predetermined time to bind DNA to the magnetic beads. When DNA is bound to the magnetic beads, the magnetic beads are trapped by a magnetic force, and the solution is discharged by a pipette. Next, the cleaning process is performed several times. In the washing step, the washing solution is added to the magnetic beads with a pipette, and the solution is stirred with the pipette to wash away unnecessary objects adhering to the magnetic beads. Then, magnetic beads are trapped by magnetic force, and unnecessary solution is discharged by a pipette. After the washing step, the DNA eluate is dispensed onto the magnetic beads with a pipette, and after dispensing, the solution is stirred with the pipette to elute the DNA adhering to the magnetic beads into the eluate. Then, magnetic beads are trapped by magnetic force, and the DNA eluate is sucked with a pipette and passed to the next step.

  Next, in the amplification step, the desired DNA is increased exponentially. As a method for efficiently amplifying a small amount of separated and purified DNA, a polymerase chain reaction (PCR method) is known. In this method, the target double-stranded DNA is thermally denatured and dissociated, two kinds of primers are annealed to the obtained single-stranded DNA, and then DNA polymerase is allowed to act to synthesize double-stranded DNA. . Furthermore, the target DNA can be amplified exponentially by repeating the cycle of primer annealing and complementary DNA extension by DNA polymerase after this double-stranded DNA is heat denatured to obtain single-stranded DNA. Is the method. The PCR method includes, for example, a step of dissociating double-stranded DNA at about 94 ° C., a step of annealing a primer at about 55 ° C., and a step of extending a complementary strand at about 72 ° C. using a heat-resistant DNA polymerase. It is performed by repeating a cycle including As an indicator of the amount of nucleic acid amplified, a label is bound to one of two types of primers for amplifying double-stranded DNA. In addition, by increasing the amount of the primer with the two types of primers, the amount of labeled single-stranded DNA (labeled target DNA) increases after amplification, and the labeled single-stranded DNA dominates. Become. As a means for amplification, a DNA extraction solution and an amplification solution are added with a pipette to a container to be heated and cooled, and the container is sealed to perform PCR.

  Next, the amplified DNA is separated and purified from other unnecessary substances. The means for separating and purifying the amplified DNA is the same as the separation and purification described in the extraction step, and detailed description thereof is omitted.

  Next, in the hybridization step, the amplified DNA is subjected to a hybridization reaction with the probe DNA of the DNA chip. Accordingly, the desired labeled DNA binds to a specific probe on the DNA chip in a specific double strand, so that the sample DNA can be specified.

  Thereafter, in the detection step, DNA labeled with a fluorescent substance or the like bound to a specific probe of the DNA chip is optically detected.

  Finally, in the result determination, the disease is determined based on the detected data.

  In the above process, contamination can occur. In a biochemical reaction using DNA as a sample, contamination generally means that other DNA is mixed into a test sample DNA to be amplified in an extraction process or an amplification process. During extraction or amplification, when adding a solution containing DNA with a pipette or stirring with a pipette, mist or splashing may occur and be mixed into a solution containing other DNA. is there. Further, when the liquid containing DNA is heated and cooled, the container is sealed, but a part of the vapor leaks and may be mixed into the liquid containing other DNA by an air flow or the like. Furthermore, there may be a case where a suspended substance containing bacterial DNA or the like that is not a test target rides on the airflow from outside the apparatus and enters a container containing another specimen. In particular, in the separation and purification after amplification in the amplification step, the solution containing the amplified DNA and having a high DNA concentration is agitated with a pipette, so that it is most affected by mist containing DNA and scattering and contamination. Is most likely to occur.

Conventionally, as means for preventing contamination, generally, scattering, mist, and steam, which are causes of contamination, are eliminated, or they are kept away from the affected area. As a conventional technique for preventing such contamination, there is a combination of shielding means and atmosphere purging means (see, for example, Patent Document 1). In this technique, mist and vapor are prevented from being transmitted from the outside by the shielding means, and are discharged from the outside by the atmosphere purge means or by flowing the mist and vapor generated in the apparatus to the outside by the purge gas. .
Japanese Patent Laid-Open No. 06-148198

  In the apparatus disclosed in Patent Document 1, since the door-shaped shielding member is manually opened and closed, when a pipette is used for handling the liquid, the shielding member for the transport opening is adapted to the pipette operation so as not to disturb the pipette transport. Need to open and close. However, a movable opening / closing type shielding member such as a shutter requires an opening / closing mechanism, which complicates the configuration. Further, when there are a plurality of pipette transport ports, a movable opening / closing type shielding member corresponding to that is required, and the pipette becomes more complicated. Furthermore, if the mist generated when the pipette dispenses liquid into the container is not purged quickly and efficiently, the mist may be mixed in the container.

  Therefore, an object of the present invention is to provide a biochemical reaction apparatus that can prevent contamination to a test sample accommodated in the apparatus without providing a complicated mechanism.

  As a result of repeated studies to solve the above problems, a biochemical reaction apparatus capable of preventing contamination with the following configuration has been found.

The biochemical reaction device of the present invention comprises at least one transport unit for transporting a substance into the device,
Purging the atmosphere in the compartment with at least one compartment formed by covering the inside of the apparatus with a shielding plate, at least one transport port provided in the compartment, a shielding member for shielding the transport port, and And an atmosphere purge means provided in the compartment, and the shielding member is provided in the transfer section, and the atmosphere purge means is configured to control the atmosphere in the compartment while shielding the compartment. A biochemical reaction device characterized by purging.

  According to the present invention, it is not necessary to drive the opening and closing of the shielding member of the transfer port, the configuration is relatively simple, and an inexpensive apparatus can be provided. The more shield members that open and close, the simpler the configuration and the more effective.

  In addition, the mist and vapor are purged quickly and efficiently in the area shielded by the shielding member during liquid handling, thereby enabling the elimination of substances that cause contamination and a highly reliable device. Can be provided.

  The conveyance part which the biochemical reaction apparatus of this invention has is equipped in order to convey a substance in an apparatus. The compartment is formed by covering at least a part of the space in the apparatus with a shielding plate, and has at least one transport port. The transport unit is provided with a shielding member that shields the transport port. By providing the shielding member in the transport unit, the shield member can shield the shielding port when the transport unit transports the substance into the compartment. The compartment is provided with an atmosphere purge means provided in the compartment for purging the atmosphere in the compartment. The atmosphere purging unit operates while the compartments are shielded, so that the compartments during the dispensing operation and the reaction process can be set in an environment in which no contamination occurs.

  As a preferred embodiment of the present biochemical reaction apparatus, an apparatus having a liquid handling unit for handling a liquid in a container installed in the apparatus can be mentioned. This apparatus has, as a transport section, a first transport section that transports a container installed in the apparatus into the apparatus, and a second transport section that transports a liquid handling section into the apparatus. And the compartment of this apparatus has the 1st, 2nd conveyance opening which can insert each of a 1st, 2nd conveyance part in a compartment. The first transport unit is provided with a first shielding member for shielding the first shielding opening, and the second transportation part is provided with a second shielding member for shielding the second shielding opening. A shielding member is provided. Further, in this apparatus, when the liquid handling unit is in a position for sucking or discharging the liquid in the container installed in the apparatus, the first shielding member is in a position for shielding the first transfer port, and the second The shielding member can be configured to be positioned so as to shield the second transport port. A pipette or the like can be used as the liquid handling unit.

  The following examples illustrate the invention in more detail.

  Hereinafter, the present invention will be described in detail with reference to examples. However, the technical scope of the present invention is not limited by these examples.

  In a DNA test apparatus to which the present invention can be applied, biochemical reactions of at least one specimen are simultaneously performed. A schematic diagram of the DNA testing apparatus is shown in FIG. 2A is a top view and FIG. 2B is a front view. This DNA testing apparatus is an apparatus that performs four process processes, an extraction process, an amplification process, a hybridization process, and a detection process. An extraction process for extracting nucleic acid from a sample, an amplification process for amplifying nucleic acid, a hybridization process for performing a hybridization reaction between a nucleic acid and a probe, a detection process for detecting a hybridization reaction, and a process for determining a result are performed in this order. .

  The process processing section includes an extraction section 1a that performs an extraction process, an amplification section 1b that performs an amplification process of the amplification process, a purification section 1c that performs a purification process of the amplification process, and a hybridization detection section 1d that performs a hybridization process and a detection process. It consists of two. In addition, a container standby unit 2 for storing a specimen, a container filled with a test reagent or reaction solution used in each processing step, and a discard unit 3 for disposal after the end of the test process are arranged. Furthermore, the hand part 11 which is a conveyance part which conveys a container to the reaction field (not shown) in each process process part and discards a container is installed. Further, a pipette unit 8 is installed, which is a combination of a pipette that is a liquid handling unit that simultaneously moves and stirs liquid between each well, and a transport unit that transports the pipette.

  As shown in FIG. 2A, the hand unit 11 is arranged on a stage that can operate in the X direction and the Y direction so that each processing unit, the container standby unit 2, and the disposal unit 3 can be accessed. The operation direction of the hand unit 11 is indicated by arrows A and B (B1, B2, B3, B4, B5, and B6).

  As shown in FIG. 2B, the pipette unit 8 is arranged on a stage that can be operated in the X direction and the Z direction so that each processing unit can be accessed and each well in the same container can be accessed. . The operation direction of the pipette unit 8 is indicated by arrows C and D (D1, D2, D3, D4).

  The extraction unit 1a, the amplification unit 1b, and the purification unit 1c are each shielded by a shielding plate. However, the hybridization detection unit 1d, the container standby unit 2, and the discard unit 3 may be shielded by a shielding plate, or only one or two process processing units may be shielded. FIG. 3 shows a perspective view of the configuration of the shielding means to which the present invention is applicable.

  The front, rear, upper, lower, left, and right six surfaces are shielded by the shielding plate 14 so as to surround the three process processing units of the extraction unit 1a, the amplification unit 1b, and the purification unit 1c. The extraction unit 1a and the amplification unit 1b are separated by the shielding plate 4a, and the amplification unit 1b and the purification unit 1c are separated by the shielding plate 4b. In this way, three compartments are formed. The extraction port 1a of the front shielding plate 14 is provided with a hand transfer port 12a, the amplification unit 1b is provided with a hand transfer port 12b, and the purification unit 1c is provided with a hand transfer port 12c. A pipette transport port 5a is provided in the extraction unit 1a of the upper shielding plate 14, a pipette transport port 5b is provided in the amplification unit 1b, and a pipette transport port 5c is provided in the purification unit 1c. As members for purging the atmosphere, an exhaust fan 9a is provided in the extraction part 1a of the shielding plate 14 on the upper surface, an exhaust fan 9b is provided in the amplification part 1b, and an exhaust fan 9c is provided in the purification part 1c. Further, intake fans 15a, 15b, 15c, and 15d are provided in the extraction portion 1a of the shielding plate 14 on the bottom surface, intake fans 15e, 15f, 15g, and 15h are provided in the amplification portion 1b, and an intake fan 15i is provided in the purification portion 1c. , 15j, 15k, 15l.

  The compartment of the process processing unit that is shielded from the pipette port and the hand port is sealed. The structure of the sealing means of the compartments of the respective process processing units is the same. After the chamber of the process processing unit is sealed, an atmosphere purge is performed to remove suspended matter containing DNA that causes contamination such as mist. The side view of the structure of the sealing means for preventing contamination and the structure of the atmosphere purge means to which the present invention can be applied is shown in FIGS. 1 (a) and 1 (b). The sealing and the atmosphere purge will be described with reference to FIG. Note that the same reference numerals as those in FIG. Reference numeral 14 denotes a shielding plate. Reference numeral 5 denotes a pipette transport port. Reference numeral 12 denotes a hand conveyance port. Reference numeral 1 denotes a process processing unit (extraction unit, amplification unit, purification unit). Reference numeral 8 denotes a pipette portion. Reference numeral 11 denotes a hand part. Reference numeral 9 denotes an exhaust fan, and reference numeral 15 denotes an intake fan.

  As shown in FIG. 1A, there is a reaction field 13 in the process processing unit 1. The shielding member 7 is attached to the pipette unit 8 and moves together with the pipette unit 8. Further, the shielding member 10 is attached to the hand unit 11 and moves together with the hand unit 11.

  As shown in FIG. 1B, in the process processing unit 1, when the hand unit 11 moves as indicated by an arrow E and the container 6 is installed in the reaction field 13, the shielding member 10 attached to the hand unit 11 is It is in a position to shield the hand conveyance port 12. Further, when the pipette part moves and operates (suction, dispensing, stirring, etc.) as indicated by the arrow F, the shielding member 7 attached to the pipette part 8 is in a position to shield the pipette transport port 5. . Further, when the pipette transport port 5 and the hand transport port 12 are shielded, the process processing unit 1 is in a sealed state. Further, the atmosphere purge can be performed by turning on the exhaust fan 9 and the intake fan 15 in a sealed state.

  FIG. 4 shows a flowchart of the sequence in the compartment of the process processing unit in the sequence of the DNA testing apparatus to which the present invention can be applied. In the flowchart of FIG. 4, the start is started when the hand unit finishes installing the container in the reaction field in the compartment of the process processing unit and at the same time the hand conveyance port is shielded. Moreover, the time when a hand part starts conveyance of a container from the compartment of a process process part to a disposal part is made into an end. The sequence of the pipette unit will be described with reference to FIG.

  First, the pipette part is moved to the position on the container where the liquid is operated (S1). At the end of the movement, the pipette transport port is also shielded, and the compartment of the process processing unit is sealed.

  Next, the atmosphere purge is turned on (S2).

  Next, a liquid operation is performed (S3). Here, liquid suction, discharge, stirring, and the like are performed.

  Next, processing unique to each process other than the liquid operation is performed (S4). For example, the extraction unit performs extraction processing, the amplification unit performs amplification processing, and the purification unit performs separation and purification processing.

  Next, the atmosphere purge is turned off (S5). At this time, since the suspended matter containing DNA that causes contamination such as mist is eliminated in the process processing section, the inside of the process processing section is cleaned.

  Next, if the liquid operation still remains in the pipette part, the process returns to S1 and S1 to S5 are repeated.

  In addition, although it was 3 compartments in this embodiment, 1 compartment, 2 compartments, 4 compartments or more may be sufficient. In the present embodiment, in addition to areas arranged in the horizontal direction, areas arranged in the vertical direction may be used. The operation may be another procedure. Further, in the pipette unit 8, the pipette that is the liquid handling unit may be allowed to move in the vertical direction even when the transport unit is in a position that shields the shielding port. Furthermore, a hand system (hand part, shielding member) or a pipette system (pipette part, shielding member) may be omitted, and manual conveyance may be performed by shielding with an openable / closable shielding member.

It is a typical side view of the compartment of the process process part of the DNA test | inspection apparatus which is embodiment of this invention. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic top view (a) and a schematic front view (b) of an overall configuration of a DNA testing apparatus according to an embodiment of the present invention. It is a typical perspective view of a structure of the shielding means of the DNA test | inspection apparatus which is embodiment of this invention. It is a flowchart of a part of process of the DNA test | inspection apparatus which is embodiment of this invention.

Explanation of symbols

1a Extraction unit 1b Amplification unit 1c Purification unit 1d Hybridization detection unit (hybrid detection unit)
2 Container standby part 3 Disposal part 4a, 4b Shield plate 5 Pipette transport port 5a-5c Pipette transport port 6 Container 7 Shield member 8 Pipette part 9 Exhaust fan 9a-9c Exhaust fan 10 Shield member 11 Hand part 12 Hand transport port 12a to 12c Hand transport port 13 Reaction field 14 Shield plate 15 Intake fan 15a to 15l Intake fan

Claims (4)

In biochemical reactors,
At least one transport section for transporting a substance into the apparatus;
At least one compartment formed by covering the inside of the device with a shielding plate;
At least one transport port provided in the compartment;
A shielding member that shields the transport port;
An atmosphere purge means provided in the compartment for purging the atmosphere in the compartment;
The shielding member is provided in the transport unit;
The biochemical reaction apparatus, wherein the atmosphere purging means purges the atmosphere in the compartment while the compartment is shielded. In biochemical reactors,
A first transport unit for transporting the container in the apparatus;
A liquid handling unit for handling liquid with respect to the container;
A second transport unit for transporting the liquid handling unit;
At least one compartment formed by covering the inside of the device with a shielding plate;
First and second transfer ports provided in the compartment;
First and second shielding members for shielding the first and second transport ports, respectively.
An atmosphere purge means provided in the compartment for purging the atmosphere in the compartment;
The first and second shielding members are provided in the first and second transport units, respectively.
The biochemical reaction apparatus according to claim 1, wherein the atmosphere purging unit purges the atmosphere in the compartment while the compartment is shielded.   When the liquid handling unit is in a position for sucking or discharging the liquid in the container, the first and second shielding members are in positions for shielding the first and second transport ports, respectively. The biochemical reaction device according to claim 2.   The biochemical reaction apparatus according to claim 2 or 3, wherein the liquid handling unit is a pipette.

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