JPH1123472A - Luminescence inspecting device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable an accurate inspection by adding a luminescent reagent into a plurality of specimen containers into which different indication reagents are introduced, detecting luminescence, determining the presence or absence of luminescence, and determining the property of a sample from the combination of luminescence. SOLUTION: In a fractionating unit 10, each fractionating pump 24 sucks a luminescent reagent from each luminescent reagent tank 28 and supplies it to a nozzle 20 via a fractionation control part 30. In a light detecting unit 12, a reagent and a DNA sample as a child specimen are introduced into each well 8, and a luminescent reagent is additionally introduced into each well 8 for inspecting the presence or absence of the reaction of an indication reagent by luminescence in each container. A photodetector 32 detects light in the wells 8 and outputs a photodetection signal 34 responsive to the amount of received light, and the results of photodetection are stored in a data processing part 38 via signal processing part 36. In a data processing unit 14, the results of photodetection in the data processing part 38 are matched with determining conditions stored in a determining condition table 40 to determine the type of the human leucocyte antigen of the specimen.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a luminescence test apparatus, and more particularly, to a human leukocyte antigen (HLA).
The present invention relates to a luminescence inspection device that automatically performs the test of n).

[0002]

2. Description of the Related Art When performing organ transplantation or the like, it is necessary that the serologic type of the donor of the organ and that of the recipient be the same. Therefore, various tests are performed on those persons. One of the tests is the HLA test.
This is for determining the type of human major histocompatibility antigen, that is, leukocyte antigen. In recent years, a test method at a gene level has been developed. Conventionally, this HLA test has been mainly performed manually.

[0003] In this test, a plurality of types of indicator reagents that specifically react with a specific gene are used. Here, the indicator reagent is a reagent that reacts with a specific base sequence in the RNA strand.

First, DNA is collected from a collected blood sample.
Is extracted, and the DNA sample is amplified by a predetermined process. The DNA sample as a parent sample is subdivided into a plurality of containers into which different indicators have been injected in advance as child samples. Next, a luminescent reagent is added to each container, the presence or absence of luminescence (positive or negative) is checked for each container, and the type of HLA is determined by comparing the combination of the luminescence with a determination table.

[0005] Here, the luminescent reagent is a reagent that emits light only for a sample that has reacted with the indicator reagent. In general, the presence or absence of luminescence is confirmed by injecting both of two types of luminescent reagents. Although different indicator reagents are injected into each container, the same two luminescent reagents are used for each container. Since the emission time due to the addition of the luminescent reagent is about several seconds, it is desirable to add the luminescent reagent immediately before the measurement of luminescence.

[0006]

However, it is very complicated and burdensome to manually dispense a luminescent reagent and match the combination of the presence or absence of luminescence with a judgment table, and the test is difficult. Takes a long time. There is another aspect that the possibility of a determination error cannot be reduced to zero. Furthermore, there is a problem that it is not possible to flexibly cope with a case where a new judgment condition is added with the development of a new indicator or the progress of an inspection method.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned conventional problems, and has as its object to eliminate the complexity and processing load during a luminescence test of a specimen and to enable an accurate test.

Another object of the present invention is to provide an apparatus capable of flexibly responding to a change in a judgment condition and checking whether the change of the judgment condition is appropriate.

[0009]

In order to achieve the above object, the present invention provides a plurality of sample containers into which a sample is dividedly injected and different indicator reagents are injected, and a luminescent reagent in each of the sample containers. A reagent supply means for adding the sample, a luminescence detecting means for detecting luminescence in each of the sample containers, and determining the presence or absence of luminescence for each of the sample containers, and determining a property of the sample from a combination of luminescence of each of the sample containers. Determining means for determining.

According to the above arrangement, the luminescent reagent is added to each sample container into which the sample and the indicator reagent have been injected,
At that time, the presence or absence of light emission in each sample container is inspected. Then, the properties of the sample are determined based on the combination of the light emission. Since a series of steps from the addition of the luminescent reagent to the determination are automatically executed, the burden on the operator can be greatly reduced, and the problem caused by human error can be solved.

In a preferred aspect of the present invention, a specific indicator reagent for generating a luminescence reference is injected into a specific sample container among the plurality of sample containers, and the determining means includes: The presence or absence of light emission of each of the other sample containers is relatively determined based on the amount of light emission of. As described above, if any one of the child samples is used as the luminescence reference, accurate luminescence determination can be performed irrespective of the sample concentration and the measurement conditions.

In a preferred aspect of the present invention, the information processing apparatus includes a judgment condition storage unit in which judgment conditions used by the judgment means are stored.

In a preferred aspect of the present invention, the apparatus further comprises a judgment condition changing means for changing a judgment condition in the judgment condition storage section. By providing means for adding, deleting, and correcting judgment conditions, it is possible to flexibly cope with various inspection methods without redesigning the system.

In a preferred aspect of the present invention, a dummy data storage means for storing dummy data prepared for verifying a new determination condition, and a dummy data based on a result of determining a property of the sample using the dummy data. Verification means for verifying whether the new determination condition is appropriate. According to this configuration, even if a new determination condition is added, it can be verified using dummy data, so that a problem due to registration of an incorrect determination condition can be avoided beforehand, and a highly reliable system can be achieved. Can be constructed.

In a preferred aspect of the present invention, the method includes a microplate having a plurality of wells formed thereon, wherein the microplate is divided into a plurality of blocks, and each parent sample is assigned to each block. . According to the above configuration, a test for a plurality of samples can be automatically performed only by setting one microplate in the apparatus, and a plurality of samples can be automatically performed by performing dispensing processing, luminescence measurement, and the like continuously. The overall test time over the specimen can be reduced.

[0016]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 shows a preferred embodiment of a luminescence inspection apparatus according to the present invention, and FIG. 1 is a conceptual diagram showing the entire configuration. This luminescence inspection apparatus performs an HLA test on human serum. That is, a leukocyte antigen test is performed to determine the type of the antigen.

In FIG. 1, the present apparatus is roughly divided into a dispensing unit 10, a light detecting unit 12, and a data processing unit 14. The system control unit 16 controls the units 10, 12, and 14.

The dispensing unit 10 uses one or a plurality of nozzles 20 to dispense two kinds of luminescent reagents. Note that the dispensing unit 10 may be used to dispense the parent sample or the indicator reagent.

The microplate 6 shown in FIG.
A plurality of well-shaped wells 8 are formed in a matrix, and each well 8 has an indicator reagent and DNA as a child sample.
The sample is injected. In this state, a luminescent reagent is additionally injected into each well in order to perform a luminescence test for the presence or absence of the reaction of the indicator reagent in each container. In this embodiment, the sample is subdivided into 24, and 24 different indicator reagents are used.

The nozzle 20 is driven three-dimensionally by a nozzle transport mechanism 22. The driving is controlled by the dispensing control unit 30. The dispensing control unit 30 controls the operation of two dispensing pumps 24 provided corresponding to the two nozzles. Specifically, the motor 26 that drives the piston shaft of the dispensing pump 24 is controlled by the dispensing control unit 30. Each dispensing pump 24 is connected to each luminescent reagent tank 2
It has an action of sucking each luminescent reagent from 8 and supplying each luminescent reagent to the nozzle 20. Note that, as in the present embodiment, the dispensing pump 24 and the nozzle 20 may be provided for each of the two luminescent reagents, but one dispensing pump 24 and the dispensing nozzle 20 are also used for those luminescent reagents. You may. Dispensing pump 24 and nozzle 20 for each luminescent reagent
Is advantageous in that cleaning of nozzles, dispensing hoses and the like becomes unnecessary. Further, such a configuration has an advantage that the time required for dispensing can be significantly reduced. As the dispensing unit 10, the configuration of a conventionally used dispensing apparatus can be used.

The light detection unit 12 includes a light detector 32 and a signal processing unit 36. In this embodiment, only one photodetector 32 is provided, and the photodetection is performed on any of the wells 8 by driving the microplate 6 in the horizontal direction by the plate transport mechanism 33. The photodetector 32 is composed of, for example, a photomultiplier tube (PMT), and outputs a photodetection signal 34 corresponding to the amount of received light. The signal processing unit 36 performs an amplification process and a conversion process to a digital signal on the light detection signal 34. The light detection result for each well is stored in a memory provided in the data processing unit 38.

Next, in the data processing unit 14, the HLA type of the specimen is determined by the data processing section 38 based on the light detection result for each well 8. Specifically, the determination condition table 40 stores a determination condition, which will be described later with reference to FIG. 4, and the HL is determined by matching a light detection result to the determination condition.
The type of A has been determined. In the present embodiment, in the determination of the type, a large classification (serologic type determination) is first performed, and a small classification is determined only when the large classification is properly performed. (Subdivision determination) has been performed.

The judgment condition changing unit 42 is a means for additionally registering a new judgment condition in the judgment condition table 40, such as a processing unit for registering a keyboard and data from the keyboard as the judgment condition. It consists of. The verification dummy data storage unit 44 is a unit that stores verification dummy data prepared to determine whether or not a determination condition additionally registered in the determination condition table 40 is appropriate. That is, by providing the dummy data for verification to the data processing unit 38, it is determined whether the additionally registered determination condition is appropriate. This will be described later in detail.

On the display unit 46, the result of the judgment made by the data processing unit 38, that is, the type of HLA is displayed while being classified into a large classification and a small classification.

FIG. 2 shows the well 8. A luminescent reagent is added to the mixture of the DNA sample and the indicator reagent injected into the well 8. As described above, the indicator reagent reacts with a specific base sequence, and the presence or absence of the reaction is detected as the presence or absence of light emission due to the addition of the luminescent reagent. As described above, light at the time of light emission is detected by the photodetector 32. It is desirable to form a light-shielding film or the like on the bottom surface or side surface of the well 8 in order to prevent crosstalk between the well and another well. In order to prevent crosstalk between wells, it is desirable to perform light detection at a position as close as possible to the upper opening of the well 8 during light detection.

FIG. 3 shows a top view of the microplate 6. As described above, a plurality of wells are formed in a matrix on the microplate 6. In the present embodiment, the plurality of wells are divided into four blocks from a first block to a fourth block. Each block corresponds to one parent sample, and in this embodiment, four samples can be measured simultaneously. Of course, the number of blocks may be appropriately set according to the number of wells 8 in the microplate 6.

In this embodiment, a single photodetector 32 is used.
However, for example, if the same number of photodetectors as the number of wells in the row direction or the column direction is used, the luminescence measurement for a plurality of child samples can be performed simultaneously.

FIG. 4 shows a plurality of judgment conditions stored in the judgment condition table 40 shown in FIG. The numbers shown on the horizontal axis indicate the numbers of the respective indicator reagents. As shown in FIG. 4, the major classification is specified by using the indicator reagents 1 to 13, and indicates the major classification of the type of HLA. The small classification is classified using the indicator reagents 1 to 23, and corresponds to the type of HLA. Incidentally, the 24 indicator reagents are used as a reference for the amount of light emission, and are usually set to always emit light. A% of the luminescence amount for the 24th indicator reagent
Those that emit light are determined to be positive, and those that emit less are determined to be negative. By providing the light emission reference in this manner, highly accurate determination with less error can be realized.

Next, the data processing unit 3 will be described with reference to FIGS.
8 will be described.

The flowchart shown in FIG. 5 is executed in a state where the light emission amount data for each well is stored in the internal memory of the data processing unit 38. In S101, each light emission amount data is obtained from the memory. In S102, 2
It is determined whether the light emission amount of the fourth sample is equal to or more than the B count. If less than B count, S
At 110, the determination is output to the display unit 46.

On the other hand, if it is determined in step S102 that the light emission amount of the 24th sample is equal to or larger than the B count,
Using A% of the light emission amount of the 24th sample as a reference value, S10
In 3, it is determined whether each indicator reagent is positive or negative with respect to the reference value.

In S104, the combination of the determination results is collated with the determination condition table 40, and first, a large classification is determined based on the result. As described above, in the determination of the large classification, the measurement results of the indicator reagents 1 to 13 are used.

If it is determined in S105 that the determination of the large classification could not be made, an inability to determine is output in S110.

In S106, the judgment of the small classification is executed based on the judgment condition table. This is the indicator reagent 1 to 23
This is performed using the measurement results up to. In S107,
It is determined whether or not the small classification has been determined. If the determination has been made, the large classification determination result and the small classification determination result are output to the display unit 46 in S108. On the other hand, if it is determined in S107 that the small classification cannot be determined, only the large classification result is output in S109.

Next, FIG. 6 shows a process when a judgment condition is added. In S201, the data processing unit 38
Thereby, a determination condition to be added is obtained. In S202, the dummy data stored in the verification dummy data storage unit 44 is used, and a process of determining a large classification and a small classification is performed on the dummy data. As a result, the judgment results of the large classification and the small classification are compared with the normal judgment results associated with the dummy data for verification. Here, the normal determination result is stored in, for example, the verification dummy data storage unit 4.
In S204 stored in S205, if the comparison result indicates that the verification result is appropriate, the process proceeds to S205.
Are added to the judgment condition table 40.

On the other hand, if it is determined in S204 that the verification result is not appropriate, the display unit 4 is determined in S206.
6, an error message is output to the display unit 46 as to whether or not to discard the determination condition in S207. Here, when the user performs an input indicating that the discard is not performed, a process for inputting a new determination condition is performed in S208, and the processes from S202 described above are repeatedly performed. On the other hand, if it is determined in S207 that the determination condition is to be discarded, the determination condition is discarded in S209.

As described above, when a new determination condition is added, the appropriateness of the determination condition can be determined by using the verification dummy data and the verification result associated therewith, so that the determination condition is erroneously added. Problems can be avoided beforehand.

[0039]

As described above, according to the present invention,
The complexity and processing load at the time of the luminescence test of the sample can be eliminated and an accurate test can be performed. Further, according to the present invention, it is possible to check whether or not the change of the determination condition is appropriate.

[Brief description of the drawings]

FIG. 1 is a conceptual diagram showing the entire configuration of a luminescence inspection apparatus according to the present invention.

FIG. 2 is a cross-sectional view showing a specific configuration of a well.

FIG. 3 is a top view of a microplate.

FIG. 4 is a diagram showing a specific configuration of a determination condition table.

FIG. 5 is a flowchart illustrating a determination process of a data processing unit.

FIG. 6 is a flow chart of a data processing unit determining whether or not an additional determination condition is appropriate;

[Explanation of symbols]

6 microplates, 8 wells, 10 dispensing units, 12 light detection units, 14 data processing units, 32 light detectors, 38 data processing units, 40 judgment condition tables, 42 judgment condition change units, 44 verification dummy data storage units .

Claims (6)

[Claims] 1. A plurality of sample containers into which a sample is subdivided and into which different indicating reagents are injected, reagent supply means for adding a luminescent reagent into each of the sample containers, and light emission in each of the sample containers. A luminescence inspection apparatus comprising: a luminescence detection unit for detecting; and a determination unit for determining the presence or absence of luminescence for each of the sample containers, and for determining a property of the sample from a combination of luminescence of each of the sample containers. 2. The device according to claim 1, wherein the specific sample container in the plurality of sample containers includes:
A specific indicator reagent for generating a luminescence reference is injected, and the determination unit relatively determines the presence or absence of luminescence of each of the other sample containers based on the luminescence amount of the specific sample container. Characteristic luminescence inspection device. 3. The light emission inspection apparatus according to claim 1, further comprising a determination condition storage unit in which a determination condition used by said determination unit is stored. 4. The light emission inspection apparatus according to claim 3, further comprising a determination condition changing unit for changing a determination condition in the determination condition storage unit. 5. The apparatus according to claim 4, wherein a dummy data storage means for storing dummy data prepared for verifying a new determination condition, and a result of determining a property of the sample using the dummy data. Verification means for verifying the suitability of the new determination condition based on the light emission inspection apparatus. 6. The apparatus according to claim 1, further comprising a microplate in which a plurality of wells are formed, wherein the microplate is divided into a plurality of blocks, and each parent sample is assigned to each block. Luminescence inspection device.