JP2012108062A - Autoanalyzer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an autoanalyzer capable of improving a processing capacity, securing reliability of data, minimizing consumption of liquid for hemolysis, for example, a pretreatment liquid and a specimen, and efficiently performing an analysis.SOLUTION: This autoanalyzer includes: a reaction cell for retaining a reaction liquid formed by reacting a specimen with a reagent; a specimen dispensing probe for dispensing, into the reaction cell, a prescribed quantity of specimen from a specimen container retaining the specimen; and an analysis section for analysing the reaction liquid of the specimen and the reagent. This autoanalyzer further includes a function for discharging, to the reaction cell, the liquid for performing a hemolytic action in the specimen dispensing probe along with the specimen when the whole blood specimen is retained in the specimen container, a prescribed quantity of the specimen is sucked by the specimen dispensing probe and then the sucked specimen is discharged to the reaction cell.

Description

  The present invention relates to an automatic analyzer for measuring the concentration or activity value of a target component in a sample containing multiple components such as blood and urine, and more particularly, to each reaction cell while sequentially transferring a plurality of reaction cells. In addition, the present invention relates to an automatic chemical analyzer that sequentially dispenses reagents and measures the absorbance of a reaction solution in the reaction cell with a spectrophotometer at predetermined time intervals. Such an automatic analyzer is also used for general chemical analysis of other samples.

  An automatic analyzer that analyzes components such as blood and urine chemically reacts a liquid sample dispensed in a reaction cell with a reagent, irradiates the reaction solution with light using a halogen lamp, etc., and measures the absorbance to measure the liquid sample. Analyze ingredients.

  Hemoglobin A1c (hereinafter abbreviated as HbA1c) is used as a diagnostic marker for diabetes, and the reference range is 4.3 to 5.8%. HbA1c has a half-life of about 30 days, and its blood concentration reflects the average blood glucose level of the preceding two months, and is important as an indicator of long-term blood glucose control.

  When measuring HbA1c (% value) with an automatic analyzer, for example, it is measured using the HbA1c reagent shown in FIG. The measurement procedure is to use a liquid for hemolysis such as purified water or pretreatment liquid to dilute blood cells or whole blood to a predetermined dilution rate, for example, 100 times, to hemolyze the sample, and to react the hemolyzed sample with the reagent. It can be measured by determining the ratio (% value) of HbA1c to Hb from the concentration of hemoglobin (hereinafter abbreviated as Hb) determined from the absorbance and the concentration of HbA1c.

  The above-mentioned hemolysis is a phenomenon caused by cell membrane destruction of erythrocytes, which can be generated by various physical, chemical and biological factors. For example, when erythrocytes are mixed with a solution with low osmotic pressure (hypertonic solution) such as purified water, extracellular water continues to flow into the cell through the cell membrane, which is a semipermeable membrane, due to the difference in osmotic pressure. A typical hemolysis phenomenon is the rupture of red blood cells.

  Here, for example, the HbA1c reagent shown in FIG. 4 uses a hemolyzed sample as a sample for serum or urine which is a sample mainly used in an automatic analyzer. To extract the serum part from the blood, the blood collection layer after blood collection (for example, a blood collection tube for biochemistry containing a coagulation promoter) is centrifuged as a pretreatment to separate the blood cell layer in the lower layer and the serum layer in the upper layer it can. The blood collection tube is set on the apparatus sample disk as it is, the serum is sucked as a sample from the serum layer of the blood collection tube supernatant by the sample dispensing probe, and the component measurement is started by dispensing the serum into the reaction container.

  On the other hand, as an example of pretreatment until the hemolyzed sample necessary for HbA1c measurement is prepared and set on the device sample disk, the blood collection tube after blood collection (for example, an EDTA blood collection tube containing an anticoagulant) is applied to a centrifuge. Separate the blood cell layer into the lower layer and the plasma layer into the upper layer. Next, a certain amount of blood cells are collected from the lower blood cell layer, and purified blood is used as a liquid for hemolysis, for example, and the blood cells are hemolyzed by dilution at a predetermined dilution rate, for example, 100 times with respect to the blood cell amount. The HbA1c measurement is started by setting the hemolyzed sample on the apparatus sample disk, sucking the sample with the sample dispensing probe, and dispensing the sample into the reaction container.

  Conventionally, the pre-treatment until the sample was hemolyzed was manually performed by the customer himself / herself, so that the treatment was complicated and took time. For this reason, there has been a demand from customers for automation of hemolysis of blood cells or whole blood samples using an automatic analyzer.

  On the other hand, in automating hemolysis treatment by an automatic analyzer, a method for preparing a hemolyzed sample must be a system that matches a measurement item using the hemolyzed sample, for example, an HbA1c measuring reagent.

  The HbA1c measurement reagent is characterized by hemolysis of blood cells used as a sample at a predetermined dilution rate (or post-hemolysis metholysis) for each reagent. If this dilution ratio is greatly different, the hemolyzed sample concentration becomes too thin or too thick, and it falls outside the measurement range of the HbA1c measurement reagent, making it impossible to measure HbA1c (% value). Also, as shown in FIG. 4, since the Hb concentration measurement reagent and the HbA1c concentration measurement reagent are separate measurement systems, for example, a hemolyzed sample for dispensing Hb concentration is dispensed into the reaction vessel No1, and the reaction vessel No2 is dispensed. Dispenses a hemolyzed sample for measurement of HbA1c concentration, analyzes in each reaction container, obtains Hb concentration and HbA1c concentration, respectively, and determines the proportion of HbA1c in Hb from the obtained Hb concentration and HbA1c concentration ( % Value). In this measurement system, the ratio between the Hb concentration and the HbA1c concentration is important. If either the Hb concentration or the HbA1c concentration value is wrong, the required HbA1c (% value) itself is incorrect. Result will be output.

  The error factors that govern instrument performance include sample dispensing systems, reagent dispensing systems, photometry systems, and washing systems. The accuracy of sample dispensing, which has a large error factor that governs performance, is particularly important, and hemolysis Dispensing accuracy of finished samples becomes important. In summary, it is necessary to lyse blood cells at a fixed dilution rate as a first point to be noted in automation. Secondly, it is necessary to dispense a certain amount of a hemolyzed sample with high accuracy into two locations for Hb concentration measurement and HbA1c concentration measurement.

  A method for automating hemolysis using an automatic analyzer was proposed based on the above two points. First, as a means for hemolyzing blood cells at a predetermined dilution rate, a reaction cell used for photometric analysis by stirring and reacting a sample and a reagent is used as a blood cell hemolysis treatment container. Blood cells are dispensed into a reaction cell using a sample dispensing probe, and a liquid for hemolysis is dispensed into the reaction cell using a reagent dispensing probe, and then stirred and hemolyzed by a stirring mechanism. As described above, by using a dispensing probe, it is possible to dispense blood cells or a liquid for hemolysis with high accuracy, so that hemolysis at a fixed dilution rate, which is the first problem, can be achieved. Secondly, means for accurately dispensing a certain amount of a hemolyzed sample into two locations for Hb concentration measurement and HbA1c concentration measurement is based on the sample from the hemolyzed sample in the reaction vessel previously used for the hemolysis treatment. This can be achieved by dispensing into reaction cells for measuring the Hb concentration and for measuring the HbA1c concentration using a dispensing probe. This makes it possible to dispense a certain amount with high accuracy, which is the second problem, and as a result, it is possible to accurately measure HbA1c (% value).

  An operation example of the automatic analyzer to which the above hemolysis treatment is applied is shown in FIG. First, after a blood collection portion 106 containing a specimen is centrifuged and sedimented from the blood cell portion 104, a predetermined amount of blood cells is dispensed into the reaction cell 211 by the sample dispensing probe 101, and then a liquid for hemolysis, for example, a pretreatment liquid is added. The blood cells are hemolyzed by dispensing and stirring by a stirring mechanism, and the hemolyzed sample is dispensed from the reaction cell to a different reaction cell, and the measurement reagent and the hemolyzed sample are reacted to react with HbA1c ( A means for measuring (% value) has been proposed (see, for example, Patent Document 1).

  On the other hand, in recent years, for example, an HbA1c reagent having a measurement procedure shown in FIG. 5 has been put on the market as an HbA1c measurement reagent for an automatic analyzer.

  First, according to the instruction of the HbA1c reagent in FIG. 5, a predetermined amount of blood cells and a liquid for hemolysis are dispensed into the reaction cell and stirred. For example, a pretreatment liquid (blood cell: 6 μL, pretreatment liquid: 150 μL) diluted 26 times with respect to blood cells is dispensed into a reaction container, and then stirred to cause hemolysis by a stirring mechanism. Next, the hemolyzed sample is dispensed into another reaction cell in an amount necessary for HbA1c reagent measurement, for example, 8 μL, and the first reagent is dispensed to perform the first reaction (Hb concentration measurement), for example, 120 μL. Stir and react to determine the Hb concentration. Subsequently, in order to make the second reaction (HbA1c concentration measurement) as it is in the reaction solution after the completion of the first reaction, the second reagent is dispensed, for example, 40 μL, stirred and reacted to obtain the HbA1c concentration. This is a procedure for obtaining HbA1c (% value) from the obtained two concentrations (Hb concentration and HbA1c concentration). As described above, the characteristic of the measurement method shown in FIG. 5 is that the Hb concentration can be obtained with the first reagent and the HbA1c concentration can be obtained with the second reagent. Compared to the above, there are various advantages in terms of rapid measurement and running cost, such as fewer types of reagents, simplification of measurement, and dispensing of samples in one time. A characteristic point to be seen from the apparatus side is that the hemolyzed sample for measuring the Hb concentration and the HbA1c concentration can be measured using the same sample as shown in FIG. Even if there is variation in sample dispensing accuracy, HbA1c (% value) can be measured with high accuracy. In other words, since the HbA1c (% value) is obtained from the ratio of the Hb concentration and the HbA1c concentration, even if the concentration in which the Hb concentration and the HbA1c concentration vary due to the difference in the amount of the hemolyzed sample, the concentration value also has the same ratio. Since it is different, HbA1c (% value) required as a result can be obtained with high accuracy.

Japanese Patent No. 3762212

  However, in the automated method of the prior art, the amount of hemolyzed sample necessary for measuring HbA1c (% value) from the hemolyzed sample as the second stage, followed by the hemolysis treatment step of the blood cell or whole blood sample as the first stage. Since there were two steps of re-dispensing to another reaction cell by the amount, the apparatus processing capacity was reduced.

  In addition, in order to measure HbA1c (% value) by the above hemolysis process, in the first hemolysis process step, blood cells or whole blood exceeding the sample amount (blood cells or whole blood) originally required for the measurement are transferred to the apparatus reaction cell. The blood cell components attached to the reaction vessel cannot be removed by the reaction cell cleaning that is automatically performed after the measurement, and the blood cell components remaining in the reaction cell adversely affect the measured value of the next item. It was sometimes affected. The reaction cell used for hemolysis treatment to avoid this adverse effect (for example, 6 μL blood cells necessary for hemolysis treatment are dispensed into the reaction cell, but the amount of blood cells necessary for actual HbA1c measurement is In contrast, when the cleaning is performed by the reaction cell special cleaning function, the processing capacity is further reduced. In addition, when a liquid for hemolysis, such as a pretreatment liquid, is used, the amount of blood cells used in the first stage of the hemolysis treatment step is large, so the pretreatment liquid is used in an amount greater than that required for HbA1c (% value) measurement. As a result, there was an extra running cost.

  Furthermore, it is important to reduce the amount of blood cells as samples in order to reduce the burden on patients during sample collection and to reduce ecological contaminants.

  Currently, automatic analyzers have a problem of reducing the amount of reaction solution to reduce the reagent consumption per item for the purpose of reducing running costs. In order to reduce the reaction volume while maintaining the current reagent to sample ratio, it is necessary to reduce the volume of the sample. Current automatic analyzer technology requires accurate sample volume. As an amount that can be secured, for example, the minimum sample dispensing amount is 1.5 μL. In other words, the accuracy of the sample dispensing amount of less than 1.5 μL cannot normally be set on the apparatus side because it cannot satisfy the specifications required for the performance of the automatic analyzer.

  Based on this technical background, hemolysis using a conventional automatic analyzer can ensure the accuracy of blood cells at the bottom of a blood collection tube using a sample dispensing probe, for example, in order to hemolyze a sample. Dispense into a reaction cell in a set volume of 5 μL or more, and use a reagent dispensing probe in it to prepare a liquid (purified water or pretreatment liquid) for hemolysis at a set volume that can ensure the same accuracy. Etc.) was dispensed at a predetermined dilution rate, for example, 26-fold dilution (eg, 6 μL of blood cells, 150 μL of pretreatment solution), and then hemolysis was performed by stirring with a stirring mechanism. That is, in order to measure HbA1c (% value), it is necessary to dilute the blood cell and the liquid for hemolysis at a dilution rate determined so that the obtained Hb concentration falls within the measurement concentration range. A series of operations to perform the analysis, that is, blood cells are dispensed into the reaction cell with a sample dispensing probe with accuracy, and a liquid for hemolysis with a reagent dispensing probe with accuracy is prepared. The mixture was dispensed and hemolyzed by stirring with a stirring mechanism for stirring the reaction solution.

  On the other hand, as a HbA1c measuring reagent for automatic analyzers in recent years, a reagent having a measuring method as shown in FIG. Characteristically, the Hb concentration can be obtained with the first reagent and the HbA1c concentration can be obtained with the second reagent. Since the sample is measured using the same hemolyzed sample, the hemolyzed sample is separated within the measurement range of the reagent. Even if there is a variation in the injection accuracy, the obtained HbA1c (% value) can be measured with high accuracy. In other words, since HbA1c (% value) is obtained from the ratio of the Hb concentration and the HbA1c concentration, even if the concentration values at which the Hb concentration and the HbA1c concentration vary due to different amounts of the hemolyzed sample, they vary at the same ratio. Therefore, HbA1c (% value) obtained as a result can be obtained with high accuracy.

  The idea of a conventional automatic analyzer is that, in principle, the relationship between a sample, a reagent, and a measurement result is to measure one measurement result from one sample using one system (for example, two reagent systems).

  Error factors that govern the performance of the instrument include sample dispensing systems, reagent dispensing systems, photometry systems, and washing systems. The accuracy of sample dispensing, which has a large error factor that governs performance, is particularly important. Can be set only within a range where accuracy can be ensured. However, the HbA1c reagent in FIG. 5 described above is characterized in that one concentration of reagent is used from one sample and two concentrations (Hb concentration and HbA1c concentration) are obtained, and one measurement result (HbA1c%) is obtained from the ratio. Even in a region where accuracy of sample dispensing accuracy of less than 1.5 μL cannot be ensured, it can be said that HbA1c% can be obtained with high accuracy.

  An object of the present invention is to provide an automatic analyzer capable of improving the processing capacity, ensuring the reliability of data, and efficiently performing analysis while minimizing the consumption of pretreatment liquid and sample for hemolysis. With the goal.

  The configuration of the present invention for achieving the above object is as follows.

  A reaction cell that holds a reaction solution in which a sample and a reagent are reacted, a sample dispensing probe that dispenses a predetermined amount of sample from a sample container that holds the sample into the reaction cell, and a reaction solution of the sample and the reagent are analyzed In an automatic analyzer equipped with an analysis unit, when a whole blood sample is held in the sample container, a predetermined amount of sample is sucked by the sample dispensing probe and then the sucked sample is discharged into the reaction cell. And an automatic analyzer having a function of discharging a liquid that performs hemolysis in the sample dispensing probe together with the sample to the reaction cell.

  With the sample dispensing probe, aspirate blood cells from the bottom of the blood collection tube for the amount necessary for HbA1c reagent measurement, and after removing the blood cells adhering to the outer wall by washing the probe outer wall with a washing solution after aspirating the blood cells After reducing the accuracy drop due to sample introduction, the sample dispensing probe after blood cell aspiration is moved to the bottom of the reaction cell, and the blood force is pushed out by the system water (purified water) in the probe or the liquid for hemolysis. Then, the blood cell and the liquid for hemolysis are agitated and subjected to hemolysis treatment, so that the required amount of the hemolyzed sample to the required concentration is dispensed by the sample dispensing probe once in the reaction cell used for HbA1c measurement. Since it is completed by the operation, the hemolysis process consisting of the conventional two steps is reduced to one step without changing the hardware configuration of the conventional automatic analyzer. It is possible to improve the management capacity. Further, since the reaction cell used only for hemolysis is not necessary, an adverse effect on the next item due to insufficient washing of blood cells adhering to the reaction cell can be eliminated, which can contribute to data reliability. Furthermore, blood cells for use in hemolysis can be reduced to the amount required for HbA1c reagent measurement only, and purified water or pre-dilution solution required for hemolysis can be minimized. The burden, environmental considerations, and running costs can be reduced.

  In addition, the setting of the hemolysis process by the automatic analyzer can be configured such that either the conventional hemolysis process by two processes or the hemolysis process by one process of the present invention can be selected according to the reagent measurement method. .

  It is possible to provide an automatic analyzer capable of improving the processing capability, ensuring the reliability of data, minimizing the consumption of liquid for hemolysis, for example, pretreatment liquid and sample, and efficiently performing analysis.

It is the figure which showed the hemolysis processing means of the automatic analyzer by this invention in the Example of this invention. It is a block diagram of the automatic analyzer in the Example of this invention. FIG. 3 is a partially enlarged view of the sample dispensing probe in FIG. 2 and its surroundings. It is the figure which showed the measuring method of the conventional HbA1c measuring reagent in the Example of this invention. It is the figure which showed the measuring method of the recent HbA1c measuring reagent in the Example of this invention. It is the figure which showed the hemolysis processing means by the conventional automatic analyzer in the Example of this invention. It is the figure which showed the result of having simulated the measurement result of the HbA1c measuring reagent in the Example of this invention according to conditions. It is a flowchart of the hemolysis processing means in the Example of this invention.

  Hereinafter, an automatic analyzer according to the present invention will be described with reference to the drawings. FIG. 2 shows a configuration example of an automatic analyzer according to an embodiment of the present invention. A sample disk 201 for setting a container containing a liquid sample, a sample dispensing mechanism 204 for partially collecting (dispensing) a liquid sample from a container set on the sample disk 201, and a reaction cell 211 for reacting the liquid sample with a reagent. And a reaction disk 206 serving as a holder thereof, reagent disks 202 and 210 for setting reagents according to measurement items, reagent dispensing mechanisms 203 and 209 for dispensing reagents, and liquid samples dispensed into the reaction cell 211 In order to stabilize the reaction of the added reagent, a stirring mechanism 208 that stirs the reaction solution, a cleaning mechanism 207 that sucks and cleans waste liquid in the reaction cell, and a control unit for these mechanisms and analysis. The set of liquid samples may be based on a sample rack instead of the sample disk. As for the reagent dispensing moving mechanism, the reagent probe may be moved in a plane by an XY mechanism instead of rotational movement.

  A conventional hemolysis treatment method in the automatic analyzer having the above configuration will be briefly described with reference to FIG. 3 which is an enlarged view of a sample dispensing mechanism and FIG. 6 which is an explanatory diagram for explaining the hemolysis treatment.

  FIG. 6 is a diagram showing a conventional blood cell component hemolyzing process. As shown in FIG. 3, there are two possible positions for suction / discharge in the reaction cell 211 of the sample dispensing mechanism 204, in addition to the normal sample discharge position 305, a hemolyzed sample suction position 304. In the sequence of the hemolysis treatment process, the blood collection tube after blood collection is first separated into a plasma portion 105 and a blood cell portion 104 by centrifugation, and then the blood collection tube 106 is set on the sample disk 201 and the sample dispensing mechanism 204 Blood cells are aspirated from the blood cell portion 104 at the bottom of the blood collection tube, and dispensed into the normal sample discharge position 305 on the reaction cell 211. Subsequently, the reaction disk 206 is rotated and moved at the reagent discharge position by the reagent dispensing mechanism 203 by adding a liquid for hemolysis, such as system water or a pretreatment liquid dedicated to the measurement reagent, and stirring by the stirring mechanism 208 to remove blood cells. Hemolyze to make a hemolyzed sample. In this embodiment, since the rotation of the reaction disk is advanced by 1/4 turn + 1 position in one cycle, the reaction cell 211 in which the hemolyzed sample is dispensed in FIG. 2 advances by 1 turn + 4 position after 4 cycles. Stop at the hemolyzed sample suction position 304. During this stop, the hemolyzed sample is aspirated by the sample dispensing probe, and the hemolyzed sample is dispensed into the empty reaction cell 211 at the normal sample discharge position 305 by the amount used for analysis. Thereafter, the reagent for Hb measurement and the reagent for HbA1c measurement that have entered the reaction cell 211 are dispensed by the reagent dispensing mechanisms 203 and 209, stirred by the stirring mechanism 208, and the reaction solution is measured by the photometer 205. The concentration is obtained, and the obtained HbA1c (% value) is calculated from the ratio of the obtained Hb concentration and the HbA1c concentration.

  On the other hand, as a performance condition required for an automatic analyzer, the accuracy tolerance limit (%) is less than or equal to 1/2 of the CV of the physiological variation within an individual (a coefficient of variation representing the accuracy of measurement). ). Therefore, in order to satisfy the tolerance limit (%) of precision, the automatic analyzer accumulates each error such as sample dispensing system, reagent dispensing system, photometric system, washing system, which is a factor governing performance, The device is designed so that the total error of the device falls below the precision tolerance limit (%). In particular, sample dispensing precision with a large error factor that governs performance is designed to be within CV1%, for example, and if it exceeds that, it will not satisfy the tolerance limit (%) of precision. Will come out. Therefore, the apparatus is normally protected so that parameters cannot be set so that dispensing cannot be performed in a region where the sample dispensing amount error technically exceeds 1% CV, for example, in a minute dispensing region of less than 1.5 μL.

  Here, when a sample hemolyzed by hemolysis by two conventional processes using an automatic analyzer is measured using the HbA1c reagent shown in FIG. I tried to simulate it.

The result of Condition 1 in FIG. 7 is the simulation result in this case. For example, it is assumed that the analysis parameter for obtaining the Hb concentration is a one-end point method with a hemolyzed sample dispensing amount: 10 μL and a first reagent dispensing amount: 150 μL. Assuming that the error of the sample dispensing volume of 10 μL is CV 1% and the total error of the device including the sample dispensing volume error is CV 1.5%, the measured value sample of the Hb (g / dL) concentration is N by statistical processing. = 20 was calculated. Similarly, it is assumed that the analysis parameters for obtaining the HbA1c concentration are a two-point end method with a hemolyzed sample dispensing amount: 7 μL, a first reagent dispensing amount: 180 μL, and a second reagent dispensing amount: 40 μL. Sample dispensing amount: Assuming that the error of 7 μL is CV1% and the total error including the sample dispensing error is CV1.5%, the measured value sample of HbA1c (g / dL) concentration is statistically processed N = 20 was calculated. HbA1c (%) was determined from the formula (1), which is a conversion formula from the determined concentration to JDS glycohemoglobin standardized HbA1c (%), and the accuracy was checked.
HbA1c (%) = 0.7 × HbA1c (g / dL)
/ Hb (g / dL) × 100 + 2.6 (1)

  As a result, as shown in Condition 1 of FIG. 7, the calculated HbA1c (% value) is an average value of 5.62 (% value), Range 0.2 (% value), and CV 0.88 (%), and HbA1c (% The accuracy of the value) was as good as 0.2% of the range with respect to the reference range of 4.3 to 5.8%, and it was confirmed that clinically usable performance was obtained.

  In contrast to this result, the results of similar simulation of what the measurement results will be when the HbA1c measurement reagent shown in FIG. 4 is used with an automatic analyzer with a large sample dispensing error. Is shown as Condition 2 in FIG. For example, assuming that the error of the sample dispensing amount of 10 μL for obtaining the Hb concentration is 9% CV, and the total error including the sample dispensing amount error is CV 10%, the sample of the measured value of Hb (g / dL) concentration N = 20 was calculated by statistical processing. Similarly, the measurement of the HbA1c (g / dL) concentration is performed on the assumption that the error of the sample dispensing amount 7 μL for obtaining the HbA1c concentration is CV 9%, and the total apparatus error including the sample dispensing error is CV 10%. N = 20 value samples were calculated by statistical processing. HbA1c (%) was determined from the formula (1), which is a conversion formula from the determined concentration to JDS glycohemoglobin standardized HbA1c (%), and the accuracy was checked.

  As a result, as shown in condition 2 of FIG. 7, the calculated HbA1c (% value) is an average value of 5.70 (% value), Range 1.1 (% value), and CV 4.99 (%), and HbA1c (% The accuracy of the (value) was as large as 1.1% of the range with respect to the reference range of 4.3 to 5.8%, indicating that it was not at a clinically usable level.

On the other hand, if an automatic analyzer having a large sample dispensing amount error is used and measurement is performed using the HbA1c measurement reagent shown in FIG. This is shown in Condition 3 below. For example, it is assumed that the analysis parameter for obtaining the Hb concentration is a one-end point method with a hemolyzed sample dispensing amount: 8 μL and a first reagent dispensing amount: 120 μL. Assume that the analysis parameters for obtaining the HbA1c concentration are a two-point end method with a hemolyzed sample dispensing amount: 8 μL, a first reagent dispensing amount: 120 μL, and a second reagent dispensing amount: 40 μL. In the case of the reagent of this example, as shown in FIG. 5, a common hemolyzed sample is used to determine the Hb concentration by the first reagent dispensing, and subsequently the second reagent is put therein to determine the HbA1c concentration. . Therefore, if attention is paid only to the sample dispensing amount as a confirmation of precision, even if the dispensing amount varies for each sample dispensing, as a result, the Hb concentration and the HbA1c concentration also vary at the same ratio. Therefore, as a simulation method, the sample dispensed amount: 8 μL of the error is CV 10%, and the measured value sample is N by statistical processing so that the Hb concentration (μmol / L) and the HbA1c concentration (μmol / L) vary at the same ratio. = 20 was calculated. Furthermore, the measured value sample was again calculated by statistical processing as an error CV of 0.5% other than the sample dispensing amount error for each calculated result. HbA1c (%) was calculated from the formula (2), which is a conversion formula from the determined concentration to JDS glycohemoglobin standardized HbA1c (%), and the accuracy was checked.
HbA1c (%) = 96.3 × HbA1c (μmol / L)
/ Hb (μmol / L) +1.62 Formula (2)

  As a result, as shown in condition 3 in FIG. 7, the calculated HbA1c (% value) is an average value of 5.32 (% value), Range 0.2 (% value), and CV 0.83 (%), and HbA1c (% The accuracy of the value) was as good as 0.2% of the range with respect to the reference range of 4.3 to 5.8%, and it was confirmed that clinically usable performance was obtained.

  As described above, when the sample dispensing amount error is large, the tolerance of precision (%) cannot be satisfied with a normal measurement reagent and cannot be used clinically. In the case of the HbA1c reagent, the accuracy of the HbA1c (% value) required clinically is good even if there is a large variation in the accuracy of the Hb concentration and the HbA1c concentration. It will be. In other words, in order to ensure the accuracy of the dispensing accuracy of the hemolyzed sample, the hemolysis treatment by the conventional automatic analyzer was performed in two steps, but if the HbA1c reagent shown in FIG. Therefore, it can be said that it can be applied if a new hemolysis treatment means that cannot secure the sample dispensing accuracy can be invented. Therefore, in the invention of the new hemolysis treatment means, it can be said that an area in which the sample dispensing amount error of the automatic analyzer is technically over 1% of CV, for example, a minute dispensing area of less than 1.5 μL can be used as an option.

  Next, a new hemolysis method for blood cells will be examined.

  The HbA1c reagent shown in FIG. 5 requires 8 μL of hemolyzed sample, and the dilution rate for lysing blood cells is 26 times. Therefore, in the hemolysis treatment by the two steps of the conventional automatic analyzer, After dispensing 6 μL of blood cells with an injection probe and a liquid for hemolysis, for example, a pretreatment solution with a reagent dispensing probe, into a 150 μL reaction cell, the mixture was stirred by a stirring mechanism to prepare a 26-fold diluted hemolyzed sample, 8 μL of the measured hemolyzed sample was dispensed into the measurement reaction cell using the sample dispensing probe. Stirring of the liquid mixture in the reaction cell is performed by, for example, rotation of a stirring bar with a stirring function, so it is necessary to mix the liquids well without causing interference between the stirring bar and the bottom of the reaction cell. And cannot be physically stirred. Due to such a background, the reason why the total volume of 156 μL of 6 μL of blood cells and 150 μL of pretreatment liquid for preparing a 26-fold diluted hemolyzed sample is due to the stirring performance.

  Here, a new hemolysis treatment means that is desired to be carried out by the automatic analyzer when measuring the HbA1c reagent is a small number of steps compared to the hemolysis treatment of the conventional automatic analyzer, and a 26-fold diluted hemolyzed blood sample necessary as a measurement sample amount The purpose is to dispense 8 μL. That is, it is to achieve the hemolysis by dispensing and stirring the blood cell 0.3 μL and the pretreatment liquid 7.7 μL in a few steps. If it can be realized, there will be merits in various points such as improved processing capability, improved reliability by reducing blood cell stains on reaction cells, and reduced running costs by reducing pretreatment liquid consumption.

  The sample dispensing probe is normally filled with system water, and when the syringe for the sample dispensing probe is driven by a set amount, the sample is sucked and discharged from the tip of the sample dispensing probe using the system water as a medium. . Therefore, if the system water used as a medium is hemolyzed with a blood cell as a liquid for hemolysis, a necessary amount of blood cells and a liquid for hemolysis can be dispensed simultaneously. In addition, due to the force of pushing out the system water together with the blood cells at the tip of the sample dispensing probe, the blood cells and the system water, which is a liquid for hemolysis, are simultaneously agitated to form two birds with one stone. System water is used as an example of the hemolyzing liquid, but if the pretreatment liquid is aspirated in advance using the sample dispensing probe before the blood cells are aspirated, the pretreatment liquid is added together with the blood cells. If extruded, dispensing and stirring are possible in the same manner. In addition, the sample dispensing amount error at the time of 0.3 μL dispensing becomes a problem. As described above, even if the accuracy of the 0.3 μL blood cell dispensing amount error is 10% CV, Condition 3 in FIG. As shown in the simulation results, there is no clinical problem with the HbA1c reagent shown in FIG. Further, even if the measurement range of the Hb concentration of the HbA1c reagent shown in FIG. 5 is, for example, 80 to 350 (μmol / L), the Hb concentration is less than 80 μmol / L as in the simulation result of condition 3 in FIG. Since this does not occur, the problem with the dilution rate of blood cells can be cleared.

  Thus, if the reagent is based on a measurement principle such as the HbA1c reagent shown in FIG. 5, the hemolysis treatment using the sample dispensing probe is performed alone and the hemolysis sample used for measurement is dispensed once. It can be completed by the sample dispensing operation.

  As an example of the present invention, FIG. 1 shows an example in which the new hemolysis treatment means described above is incorporated into an automatic analyzer.

  First, segmental air 102 is sucked in order to prevent contact between blood cells as a sample, sample dispensing probe 101 and system water 103 in the flow path. Next, when the sample dispensing mechanism 204 rotates and descends on the blood collection tube 106 which is a sample container and the tip of the probe 103 comes into contact with the plasma layer 105 by the liquid level detection function, the liquid level height 107 (the amount of blood collected) After confirming (height), the tip of the sample dispensing probe 101 rushes and stops from the bottom of the blood collection tube, which is a blood cell layer, to a height of several millimeters, for example, 2 mm, and accurately aspirates the amount of blood cell 108 used for measurement. . If the liquid level is too high, the probe tip will not reach the blood cell layer up to 2 mm in the blood collection tube bottom, so the suction operation will be stopped and an alarm will be given to inform you that the sample volume (blood collection volume) is high. Also good. In addition, when the measurement sample is not the blood cells in the lower layer of the blood collection tube but the whole blood in which the blood collection tube is mixed by inversion, the entire blood collection sample is whole blood, so there is no need to push the probe to the bottom of the blood collection tube Since the probe outer wall is unnecessarily soiled, the tip of the probe may be immersed in the sample several millimeters and stopped by the liquid level detection function, which is the same operation as when dispensing serum, and the whole blood sample may be aspirated . Thereafter, the sample dispensing probe rises, and before dispensing blood cells into the reaction cell 211, the nozzle outer wall is washed with the washing water 110 in the washing tank 301 to wash away the carry-over portion 109 of the blood cells. The removal of the sample attached to the outer wall after suction, such as a blood cell sample, may be performed by jetting a gas such as air or suction by a vacuum pump instead of the water cleaning 110 in the cleaning tank 301. Alternatively, after the water cleaning 110 in the cleaning tank 301, for the purpose of removing water droplets attached to the outer wall, a jet of gas such as air or suction by a vacuum pump may be performed.

  After that, by rotating and moving onto the reaction cell 211 and descending to the bottom of the reaction cell, the blood cells are discharged while being pushed out by the system water 103, thereby stirring the blood cells and the system water which is purified water, here purified water. Blood cells are hemolyzed by differences in osmotic pressure. Extrusion of the liquid sample at the time of discharge may be performed by a pretreatment liquid for sample hemolysis or methification designated for each reagent, instead of system water (purified water). Before aspirating blood cells, etc., with the sample dispensing probe, the required amount of pretreatment liquid is aspirated from the pretreatment liquid container installed on the sample dispensing probe trajectory in advance, and the sample is put into the reaction cell. Blood cells or the like may be discharged and stirred while extruding the pretreatment liquid during dispensing. Instead of dispensing the first reagent for measuring the HbA1c reagent immediately after dispensing a sample such as a blood cell, the first reagent may be dispensed after a certain time in consideration of the time required for hemolysis. Alternatively, in the automatic analyzer configured as shown in FIG. 2, the reagent dispensing mechanism 203 is R1 (first reagent) and R4 (fourth reagent), and the reagent dispensing mechanism 209 is R2 (second reagent) and R3 (third reagent). If the apparatus discharges the HbA1c reagent at the timing of R2, the second reagent for HbA1c reagent measurement is dispensed at the timing of R3. Using the time until the dispensing timing, the time required for hemolysis or metholysis can be secured, and the HbA1c reagent measurement is also possible.

  An example of control when the hemolysis process according to the present invention is applied to an automatic analyzer is shown together with the flowchart of FIG.

  Enter the measurement request item and check whether it is a hemolyzed sample measurement item. For example, it is determined whether it is an item for obtaining HbA1c (% value) (step S801). If it is an item for obtaining HbA1c (% value), it is selected whether or not to perform hemolysis with an automatic analyzer (step S803). If HbA1c (% value) is not an item to be calculated (No in step S801), or if the operator has already performed hemolysis of the sample using the technique, it is desired to set the hemolyzed sample in the apparatus and perform only the analysis (step 803). Therefore, the analysis of the requested measurement item is performed (step S807) without performing the hemolysis process by the automatic analyzer (step S802).

  In order to cause the apparatus to recognize that the measurement sample has been hemolyzed by a method, for example, the operator may recognize the sample state “sample hemolyzed” on the measurement item request screen. Next, when hemolysis by the automatic analyzer is necessary (Yes in S803), the water extrusion type hemolysis by the sample dispensing probe according to the present invention is performed, or the conventional two-step hemolysis is performed. Is selected (step S804). For example, a hemolysis method can be selected and registered in analysis parameters set for each analysis item. Thereby, it is possible to select and set the hemolysis treatment method for each analysis item in advance. Alternatively, the hemolysis processing means may be selected each time on the measurement request screen. Here, the necessity of the conventional hemolysis treatment by two steps is that the dilution ratio of the liquid for hemolysis with blood cells is 100 times depending on the reagent, and if the sample amount for measurement after hemolysis is 5 μL, The amount of blood cells is 0.05 μL, the amount of liquid to be lysed is 4.95 μL, the amount of blood cells dispensed is extremely small, and the precision is too far too far away from the measurement range of the Hb concentration. Even in the HbA1c measurement reagent shown in FIG. 5, the influence on the precision of HbA1c (%) cannot be wiped off. In such a case, the conventional hemolysis treatment by two steps may be selected. In addition, the initial measurement was performed by the water extrusion type hemolysis means using the sample dispensing probe according to the present invention. However, when re-examination is necessary due to an abnormality in the measured value, the conventional two steps that can be performed more accurately than the present invention means. Hemolysis treatment means may be selected. In addition, the automatic reexamination function allows the hemolysis sample item to be selected for reexamination due to the occurrence of an alarm so that the hemolysis process means at the time of reexamination can be selected in advance using the conventional two-step hemolysis means or analysis parameters. May be.

  After the hemolysis processing by the hemolysis processing means according to the present invention (step S806) or the conventional two-step hemolysis processing means (step S805), the measurement items are analyzed and the result is output (step S807).

  In this way, by setting the two hemolysis treatment means to be used properly according to the measurement method of the measurement reagent using the hemolyzed sample, it is necessary for the analysis while ensuring the accuracy of the required measurement result. An automatic analyzer capable of reducing time is provided. Use of hemolysis can be achieved by using the same dispensing probe and changing the setting of the sample aspiration amount, the system water extrusion amount, etc., with the software that controls the dispensing probe. Moreover, it is good also as a structure which provided separately the dispensing probe which optimized the shape according to the hemolysis process means.

  In addition, according to the present invention, since the reaction cell used for hemolysis is not necessary, the adverse effect on the next item when using the reaction cell due to insufficient washing of blood cells adhering to the reaction cell can be eliminated. Can also contribute to the reliability of In addition, blood cells used for hemolysis can be reduced to the amount necessary for HbA1c reagent measurement, and the amount of liquid used for hemolysis (eg, purified water or pretreatment liquid) should be minimized. Therefore, the burden on the patient, environmental considerations and running costs can be reduced.

  As described above, according to the present invention, blood cells are automatically hemolyzed using a reaction cell having a photometric function of a conventional automatic analyzer, and then a hemolyzed sample is re-dispensed to another reaction cell. Since the hemolysis process in the reaction cell used for hemolysis is omitted for items that measure blood cell components such as obtaining HbA1c (% value) by Hb concentration measurement or HbA1c concentration measurement, Improvements can be made. In addition, since the reaction cell used for hemolysis is not necessary, adverse effects on the next item when using the reaction cell due to insufficient washing of blood cells adhering to the reaction cell can be eliminated. Can contribute. In addition, blood cells used for hemolysis can be reduced to the amount necessary for HbA1c reagent measurement, and the amount of liquid used for hemolysis (eg, purified water or pretreatment liquid) should be minimized. Therefore, the burden on the patient, environmental considerations and running costs can be reduced.

101 Sample dispensing probe 102 Segmental air 103 System water 104 Blood cell layer 105 Plasma layer 106 Vacuum blood collection tube 107 Sample liquid level 108 Blood cell 109 Blood cell carryover 110 Sample water washing in sample dispensing probe washing tank 201 Sample disk 202 Reagent Disc 1
203 Reagent dispensing mechanism 1
204 Sample dispensing mechanism 205 Photometer 206 Reaction disk 207 Reaction cell cleaning mechanism 208 Stirring mechanism 209 Reagent dispensing mechanism 2
210 Reagent disc 2
211 Reaction cell 301 Sample dispensing probe cleaning tank 302 Sample disk outer periphery suction position 303 Sample disk inner periphery suction position 304 Diluted sample suction position (reaction cell)
305 Sample discharge position (reaction cell)
306 Sample dispensing probe

Claims (4)

A reaction cell that holds a reaction solution in which a sample and a reagent are reacted, a sample dispensing probe that dispenses a predetermined amount of sample from a sample container that holds the sample into the reaction cell, and a reaction solution of the sample and the reagent are analyzed In an automatic analyzer equipped with an analysis unit,
When a whole blood sample is held in the sample container, when a predetermined amount of sample is aspirated with the sample dispensing probe, the sample dispensing probe is discharged together with the sample when the aspirated sample is discharged into the reaction cell. An automatic analyzer having a function of discharging a liquid that performs hemolysis in the reaction cell. The automatic analyzer according to claim 1, wherein
A reagent dispensing probe for discharging a predetermined amount of reagent to the reaction cell is provided,
When a whole blood sample is held in the sample container, a predetermined amount of sample is sucked with the sample dispensing probe, and then the sucked sample is discharged into the reaction cell, and then into the reaction cell from which the sample has been discharged. , Having a function of discharging a liquid that performs hemolysis using the reagent dispensing probe,
An automatic analyzer comprising a selection mechanism capable of selecting whether a liquid that performs hemolysis is discharged by the sample dispensing probe or the reagent dispensing probe. The automatic analyzer according to claim 2,
The selection mechanism is a function for selecting in advance for each measurement reagent or each measurement sample whether the liquid that performs hemolysis is discharged by the sample dispensing probe or the reagent dispensing probe. An automatic analyzer characterized by comprising: The automatic analyzer according to claim 2,
The selection mechanism has a function of registering in advance, for each analysis item, whether the liquid that performs hemolysis is discharged by the sample dispensing probe or the reagent dispensing probe. Automatic analyzer characterized by

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