JP2006284324A - Micro total analysis system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a micro total analysis system allowing accurate detection with an optical detecting means. <P>SOLUTION: In the micro total analysis system, a fluid channel 6 is formed by applying fine processing, an inspection chip body is constituted by an optically transparent first substrate 22 and an optically transparent second substrate 24 stacked so as to cover the fine processed surface of the first substrate 22, mixed liquid of a specimen and a reagent is fed into the fine flow channel 6 for liquid supply partitioned between the first substrate 22 and second substrate 24, the midway of the fine flow channel is constituted as a detector 7, light is emitted to the mixed liquid having reached the detector 7, and information on an inspected object is automatically inspected with the light. An opening 24a is formed in a region of the first substrate 22 facing the detector 7, and is covered with a transparent body or a film 28 corresponding to the transparent body. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  According to the present invention, the liquid mixture in which the specimen and the reagent are mixed is guided to the detection unit, and the optical detection means arranged in the detection unit automatically detects necessary information in the liquid mixture. More specifically, the present invention relates to a micro integrated analysis system that enables detection in a detection unit with high accuracy.

In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing conventional sample preparation, chemical analysis, chemical synthesis, etc. have been miniaturized. Systems integrated on a chip have been developed.

This is a micro-TAS (Micro Total Analysis System), bioreactor, love-on
It is also called a chip (Lab-on-chips) or biochip, and its application is expected in the medical examination / diagnosis field, the environmental measurement field, the agricultural product manufacturing field, and the like.

  In particular, as seen in genetic testing, when complicated processes, skilled techniques, instrument operations, etc. are required, the test chip as an automated, accelerated and simplified micro analysis system is Since the analysis can be performed not only on the cost, the required amount of sample, and the time required, but also on any time and place, the benefit is great.

  In the field where various tests such as clinical tests are performed, the quantitativeness of analysis, the accuracy of analysis, and the economy of these test chips that produce results quickly regardless of location are regarded as important. On the other hand, since the inspection chip is severely restricted by its size and form, it is required to establish a highly reliable liquid delivery system with a simple configuration. Therefore, there is a demand for a microfluidic control element that has high accuracy and excellent reliability. The present inventors have already proposed a micropump system suitable for this (Patent Document 1). In addition, there has been proposed a micro total analysis system that enables automatic analysis of information by applying such a pump system and setting an inspection chip in an inspection apparatus.

  In the micro integrated analysis system, for example, a test chip includes a sample to be tested and a reagent for detecting information on the sample stored in separate parts in advance. Further, the sample storage unit and the reagent storage unit Are communicated with each other through a liquid passage, and a specimen and a reagent are mixed in the communication passage, and are reacted efficiently while being sent downstream.

  As described above, in the micro total analysis system configured by the inspection chip and the fluid control detection device and automatically detecting necessary information, an appropriate working fluid is led out from the fluid control detection device to the inspection chip side through the micro pump. As the working fluid is sent, the sample and the reagent are mixed, the reaction is performed in the reaction unit set to a predetermined temperature, and necessary information is detected in the detection unit by appropriate detection means. .

Here, the detection means irradiates the detection site on the analysis flow path for each inspection item with measurement light from, for example, an LED, and transmits light with an optical detection means such as a photodiode or a photomultiplier tube. Alternatively, it is a means for detecting reflected light. There are various types of optical detection means having different principles, but an ultraviolet / visible spectrophotometer is desirable.
JP 2001-322099 A

  By the way, in such a micro total analysis system that performs detection by optical detection means, it is required to perform more efficient and highly accurate detection on a test chip in which a biological substance and a reagent are stored in advance. .

  However, in the conventional inspection chip, the light irradiated from the ultraviolet / visible spectrophotometer etc. is irradiated to the inspection object through the light-transmitting resin constituting the laminate of the inspection chip. Even if it is made of plastic, reflection at the plastic interface and light passing through it are likely to become noise, and further accuracy of information has been demanded.

  The present invention has been made in view of such a situation, and in a micro total analysis system that automatically detects necessary information of an inspection object contained in an inspection chip, an optical detection means is used. Another object of the present invention is to provide a micro total analysis system that enables detection with higher accuracy.

In order to achieve the above object, the micro comprehensive analysis system according to the present invention comprises:
A fluid passage is formed by performing microfabrication, and a test chip body is configured from a light-transmissive first substrate and a second substrate laminated so as to cover the microfabricated surface of the first substrate,
A liquid mixture of a specimen and a reagent is fed into a fine channel for supplying liquid defined between the first substrate and the second substrate, and the middle of the fine channel is used as a detection unit. In the micro total analysis system configured to irradiate light to the mixed solution that has reached this detection unit, and automatically inspect the information of the inspection object by the light,
An opening is formed in a region corresponding to the detection portion of the second substrate, and the opening is covered with a transparent body or a film conforming to the transparent body.

According to the present invention having such a configuration, since a portion that prevents light irradiation does not occur in a portion that becomes an optical path of light, detection with higher accuracy can be performed.
In the present invention, at least the outer peripheral area of the opening of the second substrate is colored, and the film is thermally fused to the first substrate by performing laser irradiation on the colored area from the film side. It is characterized by doing.

If comprised in this way, sticking of a film can be performed easily.
Furthermore, the present invention is characterized in that a land for reducing the depth of the fine flow path is provided in the fine flow path constituting the detection unit.

  According to the present invention having such a configuration, for example, when a biological substance is inspected using an inspection chip, if a biotin-affinity protein such as streptadipine is adsorbed on the upper portion of the land, this biotin affinity Since the protein specifically binds to biotin labeled at the 5 'end of the primer used in the gene amplification reaction, it is possible to trap the probe labeled with biotin or the amplified gene with high sensitivity at this detection site. it can.

According to the micro comprehensive analysis system according to the present invention, highly accurate detection can be performed in the detection unit.
Further, by making the periphery of the opening colored, it is possible to perform heat fusion of the film with a laser.

  In addition, if a land is provided in the flow path constituting the detection unit, noise related to detection can be reduced as much as possible, and detection sensitivity can be improved.

Embodiments of the present invention will be described below with reference to the drawings.
FIG. 1 shows a micro total analysis system according to an embodiment of the present invention.
The micro comprehensive analysis system 10 includes an inspection chip 1 made of a single resin chip, and a fluid control detection device 2 that inspects necessary information by setting the inspection chip 1 at a predetermined position. Yes.

  For example, by injecting a gene sample extracted from blood, sputum or the like, the test chip 1 can automatically perform a gene amplification reaction by the ICAN method or the like in the chip and the detection thereof, and simultaneously perform genetic diagnosis for a plurality of genes. It is comprised as follows. The test chip 1 is a chip having a length and width of several centimeters. For example, about 2 to 3 μL of DNA is dropped into the chip and then mounted in the fluid control detection device 2 to automatically Thus, the amplification reaction and its detection can be performed.

If the structure of such a test | inspection chip 1 is shown schematically, it will be comprised as shown in FIG.
That is, the test chip 1 sends a reagent from the upstream side of the DNA amplification reagent container 4 with a working fluid, and divides the reagent into three flow paths A, B, and C, and then contains the sample container. 3a, a positive control storage portion 3b in which a positive control is stored, and a negative control storage portion 3c in which a negative control is stored. By extruding the specimen 3a, the positive control 3b, and the negative control 3c, the DNA in the specimen, the positive control, and the negative control are mixed with the DNA amplification reagent described above. Then, after amplification reaction is performed at a predetermined temperature and a predetermined time in the amplification reaction unit 11, each is divided into two and supplied to the detection unit 7. The DNA amplified by the detection unit 7 is immobilized here and stained with a specimen DNA amplification detection reagent and an internal control amplification detection reagent, respectively, and the degree of staining is determined by, for example, a light emitting element 8a such as an LED and a light receiving element 8b. Necessary information is taken out by an optical detection device made up of and the like, and the presence or absence of the target DNA is determined. In addition, the discharge part 9 which leads to the waste liquid storage part for storing the mixed waste liquid after detection is formed downstream of the detection part 7. Each liquid passage is equipped with a water-repellent valve for adjusting the timing of air venting and liquid merging, and a mechanism for discarding the leading part where the mixing state is unstable so that the liquid can flow precisely. It is configured.

FIG. 3 schematically shows the structure of the fluid detection unit 7 in the fine channel 6.
The fine flow path 6 provided in the inspection chip 1 of the present embodiment includes a first substrate 22 that has been finely processed, and a second substrate 24 that is laminated so as to cover the finely processed surface of the first substrate 22. The middle of this fine channel 6 is configured as a detection unit 7.

  Of the first substrate 22 and the second substrate 24, at least the first substrate 22 on which fine processing is performed is formed of a transparent body. In addition, a land 22 a that reduces the depth of the detection unit 7 is formed to protrude from a portion of the first substrate 22 corresponding to the detection unit 7. The land 22a may have any shape such as a rectangle, a circle, and a trapezoid. In short, the depth d of this portion should be shallower than the normal depth D of the fine flow path 6. Of course, the surface is preferably formed to be smooth.

As the material of the first substrate 22, various molding materials can be used as the material of the chip, and are used according to individual material characteristics. For example, fluorocarbon resins such as polystyrene, polyethylene, polypropylene, polyvinyl chloride, polycarbonate, polytetrafluoroethylene, polysiloxane polymers such as polydimethylsiloxane, polyolefins such as polymethyl methacrylate, polyvinyl alcohol, and ethylene-vinyl alcohol copolymer Polymers, polyester polymers such as polyethylene terephthalate and polybutylene terephthalate, polyamide polymers such as 6-nylon and 6,6-nylon, cyclic cycloolefin resins, polyarylate resins, cellulose polymers such as cellulose acetate and nitrocellulose, Examples include various inorganic glasses.

Further, when fluorescence is measured as a detection method, the fluorescence of the plastic substrate itself becomes a problem as noise. A plastic substrate that eliminates such noise has been proposed (Japanese Patent Laid-Open No. 2003-130874). Alternatively, low fluorescent plastics such as linear polyolefin, cyclic polyolefin, and fluorine-containing resin may be used.

However, the present invention is not limited to these examples.
On the other hand, the second substrate 24 can be formed of the same material as the first substrate 22, but an opening 24 a is formed in a region corresponding to the detection unit 7. The opening 24a is not limited to a shape such as a circle or a rectangle. In addition, the first substrate 22 is formed of a transparent body, whereas the second substrate may be transparent if the outer peripheral area of the opening 24a is colored. It is not limited to be colored. The colored range may be larger than the size of the opening 24a. That is, in the present embodiment, the opening 24a is covered with a transparent body or a film 28 that conforms to the transparent body, so that when the film 28 is irradiated with a laser, the irradiation light can be received and converted into heat. I need it. Therefore, the colored range may be appropriately determined according to the size of the film.

The film 28 is preferably a transparent body or a transparent body. Any resin can be used as long as it is not colored in this way.
The detection unit 7 according to the present embodiment is formed as described above.

  On the other hand, the detection device (not shown) installed in the detection unit 7 irradiates the detection site on the analysis flow path for each inspection item with measurement light from, for example, an LED, a photodiode, a photomultiplier tube, etc. This is a device for detecting transmitted light by means of optical detection. As optical detection means, there are various optical devices having different principles, but an ultraviolet / visible spectrophotometer is desirable. It may be an apparatus incorporated in the inspection device, or may be connected as a separate apparatus at the time of use.

When detecting necessary information of the inspection object supplied to the detection unit 7 by such a detection device, measurement light such as an LED passes through the transparent film 28 and the land 22a. Since the transmitted light is detected, there is little intrusion of noise, and the detection result can be improved. If biotin-binding protein such as streptavidin (avidin, streptavidin, extraavidin (R), preferably streptavidin) is applied to the land 22a in advance, biotin labeled on the probe substance or gene amplification reaction It specifically binds to biotin labeled at the 5 ′ end of the primer used in the above. Thereby, a probe labeled with biotin or an amplified gene is effectively trapped at the detection site. Thus, by providing the land 22a, the sensitivity can be improved.

As mentioned above, although the preferable Example of this invention was described, this invention is not limited to the said Example at all.
The micro comprehensive analysis system according to the present invention includes various gene analyzes, clinical examinations / diagnosis, pharmaceutical screening, pharmaceuticals, agricultural chemicals or safety / toxicity testing of various chemical substances, environmental analysis, food inspection, forensic medicine, chemistry, sake brewing. It can be used in livestock, agricultural production, agriculture and forestry.

FIG. 1 is a perspective view of a micro total analysis system according to an embodiment of the present invention. FIG. 2 is a diagram for explaining the outline of the inspection chip and the inspection method used in the micro integrated analysis system shown in FIG. FIG. 3 is a schematic cross-sectional view showing the structure of the test chip in the detection unit shown in FIG. 2, for example.

Explanation of symbols

DESCRIPTION OF SYMBOLS 2 Fluid control detection apparatus 4 Reagent storage part 6 Fine flow path 7 Fluid detection part 10 Micro comprehensive analysis system 22 1st board | substrate 22a Land 24 2nd board | substrate 24a Opening 28 Film

Claims (3)

A fluid passage is formed by performing microfabrication, and a test chip body is configured from a light-transmissive first substrate and a second substrate laminated so as to cover the microfabricated surface of the first substrate,
A liquid mixture of a specimen and a reagent is fed into a fine channel for supplying liquid defined between the first substrate and the second substrate, and the middle of the fine channel is used as a detection unit. In the micro total analysis system configured to irradiate light to the mixed solution that has reached this detection unit, and automatically inspect the information of the inspection object by the light,
A micro integrated analysis system, wherein an opening is formed in a region corresponding to the detection portion of the second substrate, and the opening is covered with a transparent body or a film conforming to the transparent body.   At least the outer peripheral area of the opening of the second substrate is colored, and the film is thermally fused to the first substrate by performing laser irradiation on the colored area from the film side. The micro total analysis system according to claim 1.   3. The micro total analysis system according to claim 1, wherein a land for reducing a depth of the micro flow channel is provided in the micro flow channel constituting the detection unit.

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