JPH045568A - Method for measuring specimen - Google Patents

Abstract

PURPOSE:To widen the concn. range of specimens with one time of processing by diluting the specimen at different magnifications and measuring the plural specimens. CONSTITUTION:A 1st dilution rate is first so set that a normal concn. range can be measured. A 2nd dilution rate is then set in such a manner that all the concn. ranges of the objects can be measured. A control section 115 reads the 1st, 2nd dilution rates out of a ROM in accordance with the assignment of the objects to be measured. A sample 103, reagent 104 and diluting liquid 106 of the 1st dilution rate are added via dispensers 105, 107 to a reaction vessel 101 and are uniformly mixed by a stirrer 114. A turn table 102 is rotated stepwise and a 2nd reagent 108 is added by a dispenser 109. The measurement of the vessel 101 is executed by an LED 110 and a photosensor 111. The measurement is more effective if the different dilution rates are so set that the measurable concn. ranges of the measuring objects are continuous or partly overlap. The specimens are surely measured with one time of measurement according to this constitution.

Description

[Detailed Description of the Invention] [Field of Industrial Application] The present invention relates to a method for measuring a specimen in the medical testing field, and in particular, to a method for measuring a specimen that can reliably measure a target object with a single specimen measurement. .

[Conventional technology]

In the medical testing field, there is a wide variety of information that can be obtained from blood. Red blood cells in the blood.

It contains not only leukocytes and platelets, but also various other antigens and antibodies, and by measuring these according to the purpose, various tests can be performed.

Conventionally, the method for testing samples such as blood has been to dilute the sample in advance at an assumed appropriate dilution ratio so that the concentration of the target substance, such as antigen or antibody, fits within the measurable range of the measuring device. A commonly used method is to perform the measurement after diluting the sample. The reason why the sample is diluted is that the measurable range of the measuring device used for the sample concentration is narrow. Generally, when measuring (or quantifying) an unknown sample, first find out the rough concentration using a predetermined method, then adjust (dilute) the sample concentration so that it falls within the measurable range of the measuring device used, and then proceed with the measurement. However, when blood or the like is used as a specimen, the possible concentration range of the specimen can be assumed as described later, and it has a predetermined distribution, so it is assumed that the specimen is suitable in advance. A method of diluting at a dilution ratio is applied. For example, when plasma is used as a sample and the amount of a specific antigen (concentration of a measurement target) in the plasma is tested, the measurable range of the measuring device used for the measurement is 0 to 5000/d1. Variation range of antibody amount in specimen (range of antibody amount that a specimen can have)
is 1000~100000■/d1. If the normal value range of antibody amount (the range of antibody amount in normal people, in other words, the range with the highest distribution frequency) is 2000 to 10000 /d1, the measurement device can be adjusted by diluting the sample to 1/10. O~50000■/d1 by measuring the measurable range of
We try to test for antibody levels in the following range. Therefore, the dilution ratio of the specimen (here, 10 times) is determined based on the above-mentioned normal value range of the antibody amount (the range with the highest distribution frequency). This makes it possible to measure a large number of specimens, and since it is not necessary to adjust the concentration for each individual specimen, the measurement can be carried out efficiently.

[Problems to be Solved by the Invention] However, according to the conventional sample measurement method, measurement efficiency can be improved by adjusting the range with the highest distribution frequency (range of normal values) to the measurable range of the measuring device. However, if the measured sample falls outside the measurable range of the measuring device, it is necessary to dilute it to an appropriate concentration and measure it again (re-testing), which takes time and effort. .

In addition, the determination of whether the sample concentration is within the measurable range of the measuring device is made by continuously monitoring the reaction progress of the sample to be measured and predicting whether or not it will fall within the measurable range.
Judgment is not always perfect, and furthermore, if the sample to be measured has an abnormally high concentration, the so-called brocine phenomenon will cause the measured value to appear as if it were a low-concentration sample, leading to the misunderstanding that the sample being measured has a low concentration. There were also problems.

The present invention has been made in view of the above, and an object of the present invention is to be able to reliably measure a specimen in a single measurement and to avoid retesting, which is laborious and time-consuming.

[Means to solve the problem]

In order to achieve the above object, the present invention dilutes the sample in advance at a dilution rate assumed to be appropriate so that the concentration of the target substance to be measured, such as an antigen or antibody, fits within the measurable range of the measuring device, and then performs the measurement. In the sample measurement method to be executed, a plurality of diluted samples are created by diluting the sample at different dilution ratios, and
The present invention provides a sample measurement method for measuring a plurality of diluted samples.

[Effect]

The sample measurement method of the present invention widens the concentration range of the sample that can be measured in one process by diluting the sample at different dilution ratios to create multiple diluted samples and measuring the multiple diluted samples. do.

〔Example〕

Hereinafter, the method for measuring a sample according to the present invention will be explained in detail.

FIGS. 1(a) and 1(b) show the configuration of a first embodiment of a measuring device to which the sample measuring method of the present invention is applied.
2, and a sample dispenser 10 that dispenses the sample 103 and the first reaction reagent 104 into the reaction container 101 stopped at a predetermined position a.
5, a diluent dispenser 107 that dispenses the diluent 106 into the reaction container 101 for dilution, a reaction reagent dispenser 109 that dispenses the second reaction reagent 108 into the reaction container 101, and a sample dispenser A LED IIO and a photosensor 111 (optical detection means) are used to measure chemical changes in the reaction vessel B101 that has been subjected to a predetermined dispensing process via the injection device 105 and the reaction reagent dispenser 109, and the LEDIIO and photosensor 111 A data processing unit 112 that inputs measured measurement data and performs arithmetic processing, a printer 113 that outputs the processing results of the data processing unit 112, and a stirring probe that stirs the solution in the reaction vessel 101 to make it uniform. 114, and a control unit 115 that controls the dispensing process of each dispenser 105, 107, 109, controls the rotation of the rotary table 102, controls the measurement of the LEDllo and photosensor 111, controls the stirring probe 114, etc. .

In the above configuration, the sample measuring method will be explained in the following order: (1) setting the dilution ratio, (2) controlling the control unit, and (2) operating the measuring device.

■Setting the dilution ratio In the first example, the sample was diluted with different dilution ratios.
prepare diluted samples, and at least one of the diluted samples.
The first is to set the measurement target to be within the measurable range of the above-mentioned measuring device when the concentration is within the normal concentration range;
The dilution ratio is set so that the measurable concentration ranges of the measurement target obtained by measuring individual diluted samples are continuous or partially overlap.

FIG. 2(a) is a diagram for explaining the setting of dilution factors for two diluted samples. For example, the measurable range of the measurement target A in the specimen by the measuring device is 0 to 100/semi, and the normal concentration range (concentration in the specimen of a normal person) is 100 to 200.
mg/d1. If the concentration range of measurement target A, including normal values and abnormal values (however, it does not necessarily fall within this range as abnormal values are included) is 100 to 5000 cm/plane, first, the normal concentration is measured with the measuring device. The first dilution factor is set so that the range can be measured. Here, 100-2
In order to measure 0 Off1g/di (normal concentration range) within a measurable range, the sample was diluted 10 times and used as sample source W.
j, concentration range of O to 1000 mg/d1
■Measure as /d1. Next, a second dilution ratio is set so that the measurement device can measure the entire concentration range of the measurement target A. As mentioned above, measurements can be made in the range of 0 to 1000 μ/dfl using the diluted sample prepared at the first dilution ratio, so here we will use at least 1000 to 5000 μ/dfl.
It is necessary to set the second dilution factor so that concentrations in the range of 1 can be measured. Therefore, the sample is diluted 50 times, and the concentration of the sample stock solution is measured using a measuring device at a concentration of 0 to 5000 .mu./d1. That is, the measurable range of the analyte A with the first dilution ratio and the second dilution ratio is as follows, and it becomes possible to measure the entire concentration range of the analyte A with two diluted samples. .

First dilution rate - 0 to 1000 µ/d Second dilution rate - 0 to 5000 mg/a According to the above dilution rate, 0 to 1000 µ/d by measurement with two diluted samples.
The ranges of d1 overlap, and this portion will be measured twice. In addition, for measurement target A, settings were made so that the entire normal concentration range could be measured at the first dilution ratio, but for example, for measurement target B (conditions other than the normal concentration range are the same as measurement target A), As shown, a portion of the normal concentration range may be measured separately for the diluted specimens at the first and second dilution ratios.

Furthermore, as shown in measurement object C, when the normal concentration range is within the measurable range of the device, the dilution ratio may be set to 1 (that is, the undiluted solution is used without dilution).

FIG. 2(b) shows an example of setting the dilution ratio in the measuring device of the present embodiment by specifically listing the object to be measured.

AFP (Alpha Fetoprotein) AFP is a substance that is produced when there is a tumor such as cancer in the body, and its normal concentration is extremely low, about 20 ng/m or less, and anything higher than that is considered abnormal.

Therefore, at the first dilution ratio (1x), O ~ 2000 ng,
/mfl is measured, and the range of O to 1 million ng/ml is measured at the second dilution factor (500 times). The concentration range of AFP is 0 to 1 million ng/m! can all be measured.

11. βz-M (β2-microglobulin) First dilution ratio (1x) 'T: 0 to 2200 ug/Q was measured, and the second dilution ratio (25x) was 0 to 60000 μg/Q.
Measure a range of 42. Although a concentration greater than 60,000 μgel cannot be measured, it can be treated as a clearly abnormal value in the output of the measuring device, so there is no practical problem.

iii, IgC, (IgG antibody) 0 to 50oo■/di at the first dilution factor (1x)! 0 to 10,000 ■/d at the second dilution rate (2x)
Measure the range of 1. In the case of IgG, the normal concentration range can be measured at both the first and second dilution ratios.

Note that the measurable range of the apparatus shown in FIGS. 2(a) and 2(b) differs depending on the apparatus used, and is therefore not limited to this range. Furthermore, since the concentration range of the measurement target also depends on the maximum and minimum values of the abnormal sample concentration, these are empirical values and are not limited thereto. Furthermore, even if the conditions are the same as those described above, it is of course possible to set a different dilution ratio.

Control of the control unit The control unit 115 controls the dispensing process of each dispenser 10'5, 107°109, controls the rotation of the rotary table 102, controls the rotation of the rotary table 102,
Although it is responsible for overall control of the measuring device, such as measurement control of the DIIO and photosensor 111, and control of the stirring probe 114, control of the dilution process will be particularly explained based on the dilution ratio, which is the main part of the present invention.

The control unit 115 stores the dilution ratio of each measurement object in its internal ROM (not shown) as default values as shown in the table below.

The table control unit 115 inputs the measurement object (
When a measurement item name) is specified, for example, when AFP is input, the sample 103 is dispensed and diluted based on the corresponding first and second dilution factors. Specifically, based on the first dilution ratio (1x), the amount of sample 103 to be dispensed and
Calculate the ratio of the amount of diluted liquid 1060 dispensed, and add the sample dispenser 10
5 to dispense the sample 103 into the reaction container 101 at a predetermined position, and then control the diluent dispenser 107 to dispense a predetermined amount of the diluent 106 to perform dilution. Since the magnification is 1x, the amount of diluent 106 actually dispensed is zero). Subsequently, when the next reaction container 101 is transported to a predetermined position, the ratio between the amount of sample 1030 dispensed and the amount of diluted liquid 106 dispensed is calculated based on the second dilution ratio (500 times). The sample dispenser 105 is controlled to dispense the sample 103 into the reaction container 101 at a predetermined position, and then the diluent dispenser 107 is controlled to dispense a predetermined amount of the diluent 106 to perform dilution. Note that the same sample 103 is used in the two dispensing and diluting processes described above.

That is, the control unit 115 controls each sample 103 to be measured at two different dilution ratios.

The first and second dilution factors stored in the ROM are as follows:
As described above, they are used as default values, but if the first and second dilution ratios are input via a predetermined input means, dilution is performed based on the input values and measurement is performed.

■Operation of the measuring device First, as a preparation process for starting the measurement, the ζ sample 103, the first reaction reagent 104. The second reaction reagent 108 and the diluent 106 are placed at predetermined positions, and the input means (
(not shown) to specify the object to be measured. Control unit 11
Based on the designation of the object to be measured in step 5, the first and second dilution factors are input from the ROM. Subsequently, when a predetermined measurement start button is pressed, the reaction container 10 stopped at the predetermined position a is
1, a sample 103 in an amount based on the first dilution factor, a first reaction reagent 104. and 1 diluent 106 are added, and the solution is mixed uniformly using the stirring probe 114 (first dispensing process). When the first dispensing process is completed, the control unit 115 rotates the rotary table 102 by one reaction container, moves the reaction container 101 from the predetermined position a to a predetermined position, and moves it to a new predetermined position a. Sample 103 and first reaction reagent 104 .in an amount based on the second dilution factor are placed in reaction container 101 . Then, the diluent 106 is added, and the solution is mixed uniformly with the stirring probe 114. On the other hand, the second reaction reagent 108 is added to the reaction container 101 at a predetermined position (the reaction container 101 to which the sample 103 etc. were added in the first dispensing process) via the reaction reagent dispenser 109 ( second dispensing process). When the second dispensing process is completed, the control unit 115 rotates the rotary table 102 and turns on the LED II.
Reaction vessel 10 in a predetermined position with O and photosensor 111
Measurement 1 is carried out, and the device is transported to a predetermined position C and stopped. At the same time, due to this rotation, the reaction container 101 at the predetermined position a is transported to a predetermined position and stopped. Thereafter, the second reaction reagent 108 is similarly added to the reaction container 101 having a predetermined value ib via the reaction reagent dispenser 109, and then measurement is performed.

FIG. 3 shows the relationship between the order in which the reaction vessel 101 stopped at a predetermined position a is counted as the first one at the start of measurement, the sample 103, and the dilution ratio.
When measuring three samples of , z, as shown in the figure, 1
Sample χ is dispensed and diluted at a first dilution rate into the second reaction vessel 101 for measurement, and sample X is dispensed and diluted at a second dilution rate into the second reaction vessel 101 for measurement. ,
Sample Y is dispensed and diluted at the first dilution rate into the third reaction vessel 101 and measured, and sample Y is dispensed and diluted at the second dilution rate into the fourth reaction vessel 101 for measurement. The sample Z is dispensed and diluted at the first dilution ratio into the fifth reaction container 101 for measurement, and the sixth reaction container 101 is
In this step, the sample Z is dispensed and diluted at the second dilution ratio and measured. In other words, one sample is measured at two different dilution ratios. In addition, these two dilution ratios are set by the method described above so that at least one of the dilution ratios can measure the normal concentration range of the measurement target, and is set so that it sufficiently covers the concentration range of the measurement target. For example, when measuring AFP, 50
■/d1 (normal value) sample or 2000■/d
Even if there is a sample with an abnormal value of 1000 .mu./d1 or more, it can be reliably measured using the method described above.

In the first embodiment, the diluent dispenser 107 is used to
Dispense the diluent 106 into the reaction container 101 and add it to the reaction container 1.
Although the configuration is such that dilution is performed within 01, the present invention is not limited to this. For example, after dispensing the sample 103 into a predetermined dilution container, diluting the sample 103 by dispensing the diluent 106 into the dilution container,
Of course, a configuration in which the diluted sample 103 is dispensed into the reaction container 101 may also be used.

FIGS. 4(a) and 4(b) show the configuration of a second embodiment of a measuring device to which the sample measuring method of the present invention is applied. ), a double reaction vessel 401 having two vessel parts 401a and 401b is used. The other configurations are the same as those in the first embodiment and are denoted by the same reference numerals, so the explanation will be omitted.

FIG. 5 is a diagram for explaining the operation when a double reaction vessel 401 is used, and shows the order in which the double reaction vessel 401 stopped at a predetermined position a at the start of measurement is counted as the first, and the sample. 103 and the dilution factor. For example, as sample 103, X, Y.

When three samples of Z are measured, as shown in the figure, sample The sample X is dispensed and diluted into the container portion 401b of the container 401 at a second dilution ratio. When the sample is dispensed into both container sections 401a and 401b, the first double reaction container 401 is measured. That is, measurements of the container parts 401a and 401b are performed simultaneously. Similarly, sample Y is dispensed and diluted at the first dilution ratio into the container section 401a of the second double reaction container 401, and sample Y is dispensed and diluted at the first dilution ratio into the container section 401b of the first double reaction container 401. The liquid is dispensed and diluted at a dilution factor of 2, and the measurements of the container portions 401a and 401b are similarly performed at the same time. Therefore, by using the dual reaction vessels 401, the number of vessels can correspond to the number of samples that can be measured. In other words, one sample can be measured at two different dilution ratios in one reaction vessel.

〔Effect of the invention〕

As explained above, in the sample measurement method of the present invention, multiple diluted samples are created by diluting the sample at different dilution ratios, and multiple diluted samples are measured. measurements can be carried out, thereby avoiding laborious and time-consuming re-examinations.

[Brief explanation of the drawing]

FIGS. 1(a) and 1(b) are explanatory diagrams showing the configuration of a first embodiment of a measuring device to which the sample measuring method of the present invention is applied, and FIG.
Figure (al is an explanatory diagram of setting the dilution ratio of two diluted samples,
FIG. 2(b) is an explanatory diagram showing an example of setting the dilution ratio in the measuring device of the first embodiment, FIG. 3 is an explanatory diagram showing the relationship between the order of reaction vessels, the sample, and the dilution ratio, and FIG. (a),
(b) is an explanatory diagram showing the configuration of a second embodiment of the measuring device to which the analyte measuring method of the present invention is applied, and FIG. 5 is an explanatory diagram showing the operation when dual reaction vessels are used. Explanation of symbols 101--Reaction container 102--Rotary table 103-1--Sample 104--First reaction reagent 105--Sample dispenser 106--Dilution liquid 1
07-------Diluent dispenser 108--Second reaction reagent 109--Reaction reagent dispenser 401 a LED 11 Data processing section Stirring probe Dual reaction container 40 l b-1 Container part photo sensor link control part

Claims (2)

[Claims] (1) In a sample measurement method in which the sample is diluted in advance at a dilution rate assumed to be appropriate so that the concentration of the target substance such as an antigen or antibody fits within the measurable range of the measuring device, the sample is measured. A method for measuring a specimen, comprising: diluting a sample at different dilution ratios to prepare a plurality of diluted specimens, and measuring the plurality of diluted specimens. (2) In claim 1, the different dilution ratios are set such that measurable concentration ranges of the measurement target obtained by measuring the plurality of diluted samples are continuous or partially overlap. Specimen measurement method.