JP2963793B2 - Body fluid component analysis method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method of reacting a body fluid sample such as blood with a reagent in a reaction container to detect a specific component contained in the sample. A method for confirming that.

[0002]

2. Description of the Related Art In an infectious disease test or a cross-compatibility test by an agglutination reaction, a biochemical sample in which a body fluid sample such as plasma or serum and a test reagent are dispensed into a reaction vessel and mixed and reacted. An analyzer is used. In this case, if the dispensing of the sample is not performed as expected, the measured value becomes abnormally low, resulting in a low value abnormality. For this reason, when a retest is required or when dispensing failure is overlooked, the sample is excluded from the group of healthy subjects. Conversely, if an infectious disease test, which is one of the pre-transfusion tests, occurs as described above, an infectious disease positive person (non-healthy person) is determined to be an infectious disease negative person (healthy person), and Which results in the propagation of
Similarly, in a cross-match test, which is one of the pre-transfusion tests, there is a possibility that the blood is determined to be compatible despite the combination of incompatible blood. For this reason, there has been a strong demand for a method capable of determining whether plasma or serum as a specimen is correctly dispensed into a reaction container.

On the other hand, as one means for confirming the dispensing of a sample at the time of blood type and infectious disease examination, Japanese Patent Application Laid-Open No.
No. 61 is known. However, this conventional technique is a confirmation method in a dilution step which is a pretreatment of a sample. That is, since the method is not a method for confirming the dispensing of a sample into a reaction container used for actual determination, the method cannot sufficiently meet the demand.

[0004]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and an object of the present invention is to cause a body fluid sample such as blood to react with a reagent in a reaction vessel, thereby obtaining a specific component contained in the sample. It is an object of the present invention to provide a method for easily confirming whether or not a body fluid sample such as blood has been correctly dispensed into a reaction container when detecting the blood sample.

[0005]

The method of component analysis of body fluids according to means and acts INVENTION The present invention includes the steps of dispensing at least a body fluid specimen into the reaction vessel in order to perform a mixing reaction with a body fluid sample and a reagent, the hydrogen of the body fluid sample Color tone depends on ion concentration
Dilute acidic or alkaline solutions with variable components
At least after the dispensing step of the body fluid sample
Adding to the reaction vessel such that a change occurs, and the color tone change data is stored in the reaction vessel while the body fluid sample is contained in the reaction vessel.
A step of obtaining in a state, a step of obtaining an analysis result corresponding to the mixing reaction, and a step of determining whether the analysis result is appropriate based on the data of the analysis result and the data of the color tone change. It is assumed that.

[0006] As a component used in the method of the present invention, the color tone of which changes depending on the hydrogen ion concentration, a pH indicator having a color changing range near neutrality is used. Such a pH indicator is not particularly limited, and for example, sodium alizarin sulfonate or cresol red can be used.

[0007]

The bodily fluid has a property that the hydrogen ion concentration (pH) is constant, that is, has a buffering ability to keep the pH at around 7.4. Due to this buffering capacity, when a relatively dilute acid solution or alkali solution and a body fluid are mixed, the mixed solution shows a pH near neutrality.

Therefore, according to the present invention, when a dilute weakly acidic or weakly alkaline diluted solution containing a pH indicator having a discoloration area near neutrality is used, the body fluid is mixed with the diluted solution alone and with the diluted solution. Sometimes the color tone changes. Therefore, it is possible to easily determine whether or not the body fluid has been correctly dispensed into the reaction container from the change in the color tone. In addition, the following specific effects can be obtained.
Wear. That is, blood cells are tested for the ABO blood group test.
Frontal test and Ura test for plasma
There is a check. In a front test, the test target is blood cells.
Blood cells are dispensed at the time of determination of agglutination
It can be easily determined whether or not it is. However, if plasma is
ABO-type blood group testing for back tests and infectious disease tests
However, it was not possible to confirm the plasma dispensing. However,
You can easily confirm the dispensing of plasma by applying
It became so. This method can be applied to both plasma and serum.
Being able is one of the advantages .

[0009]

EXAMPLES Example 1 Example using sodium alizarin sulfonate as pH indicator 1) Preparation of dilute solution

Sodium chloride (NaCl, MW; 58.44)
) And citric acid (C ₆ H ₈ O ₇ .H ₂ O, M
W; 210.14) 0.21g and, alizarin rinse sulfone sodium _{(C 14 H 7 NaO 7 S} · H 2 O, MW; 360.2
8) 20 mg was completely dissolved in 100 ml of purified water.
This solution showed a pH of 3.2 and became a yellowish solution. 500 μl of plasma is added to 1,000 μl of this solution,
Three-fold diluted plasma was prepared. At this time, the color tone of the diluted solution changed from yellow to reddish purple when plasma was added. This will be described in more detail below.

Table 1 shows that each of plasma A and plasma B
The pH change when the above-mentioned diluted solution was added to μl is shown. Table 2 shows 510 n of sodium alizarin sulfonate used as a pH indicator in the dilute solution.
The relationship between the absorbance at pH m and the pH is shown. Also,
FIG. 1 is a graph of the pH / absorbance data shown in Table 2.

[0012]

[Table 1]

[0013]

[Table 2]

From the above data, it can be seen from the above data that even when a 6-fold dilution is performed using the above-mentioned diluted solution, the pH drops only to around 6.5, so that the color degree of the diluted plasma is from 6-fold dilution to 6-fold.
It was found that there was almost no change until doubling dilution. Therefore, it is possible to determine the presence or absence of plasma by observing the degree of magenta coloration. 2) Application to cross-matching test

Donor blood cells (10 μl) and the above diluted solution (1.0)
The suspension was mixed with 00 μl to prepare a 1% blood cell suspension. After adding 40 μl of this blood cell suspension to the wells in the microplate, 10 μl of the recipient plasma was further added to the wells. Subsequently, the inside of the well was sufficiently stirred by an operation such as applying vibration. After allowing the plate to stand for about 30 minutes to form a hemagglutination image, judgment of the agglutination image and confirmation of plasma dispensing were performed. Since the solution was colored purple-red in the well into which the plasma was dispensed, the presence or absence of the plasma could be easily determined.
In addition, it was confirmed that the diluted solution had no effect on the agglutination reaction in the blood group test. Second Example In the case of using cresol red as a pH indicator

Sodium chloride (NaCl, MW; 58.44)
) 0.9 g and trishydroxymethylaminomethane [H ₂ NC (CH ₂ OH) ₃ . MW; 121.14] 0.12
g was added to 90 ml of purified water. 20 mg of cresol red (C ₂₁ H ₁₈ O ₅ S, MW; 382.44) dissolved in a small amount of ethanol was added thereto, and purified water was further added to adjust the total amount to 100 ml. As a result, a red-purple solution having a pH of 8.8 was obtained.

The plasma obtained is added to 1,000 μl of the solution obtained above.
0 μl was added to prepare 3-fold diluted plasma. At this time, the reddish purple color of the diluted solution turned yellow when the plasma was added.
This will be described in more detail below.

Table 3 shows that each of plasma A and plasma B
The pH change when the above-mentioned diluted solution was added to μl is shown. Table 4 shows the absorbance at 570 nm and pH of cresol red used as a pH indicator in the dilute solution described above.
The relationship is shown. FIG. 2 is a graph of the data in Table 4.

[0019]

[Table 3]

[0020]

[Table 4]

From the above data, it can be seen that even if the diluted solution is diluted 6 times with the above-mentioned diluted solution, the pH does not become around 8.0 or more. Is hardly observed. From the above facts, similarly to Example 1, even when the diluted solution prepared in this example is used, it can be determined whether or not plasma is contained by its coloration degree. In addition, the body fluid sample described above
Is not limited to blood typing, but also for various infectious disease tests
It can also be used for analysis results other than the agglutination reaction.
Needless to say, the present invention can be similarly applied to the case where

As described in detail above , claim 1 of the present invention
According to this, when a body fluid sample such as blood is allowed to react with a reagent in a reaction container and a specific component contained in the sample is analyzed, a dilution is performed.
Simply add a thin solution to the reaction vessel and react the body fluid sample to the reaction volume.
Reaction volume after the body fluid sample dispensing process
The body fluid sample is correct in the reaction vessel based on the color tone of the liquid in the vessel.
Since it is possible to determine whether or not the analysis result is appropriate while easily confirming whether or not the liquid has been properly dispensed, it is possible to easily prevent a serious determination error in the component analysis of the body fluid. In particular, the solution had <br/> use the Rukoto containing components of color tone change by hydrogen ion concentration as a dilute solution, as demonstrated in the above examples, even if the body fluid sample is diluted treated, reliably Since the presence of the sample in the reaction vessel can be confirmed in a short time, stable propriety determination can be performed. Also, as demonstrated in the examples,
By using a dilute solution as described above, color change
Data can be obtained without any problems even after mixing the body fluid sample and reagent
Therefore, the analysis result can be obtained as described in claim 2 of the present invention.
The state where the color change data was also obtained at the stage when the fruit was obtained
And dispensing confirmation is further facilitated.

[0023]

[Brief description of the drawings]

FIG. 1. pH of alizarin sodium sulfonate solution
And a graph showing the relationship between the absorbance at 570 nm.

FIG. 2: pH of cresol red solution and its 570 n
The graph which shows the relationship with m absorbance.

Claims (2)

(57) [Claims] A step of dispensing at least a body fluid sample into a reaction vessel to perform a mixing reaction between the body fluid sample and a reagent; and changing a color tone depending on a hydrogen ion concentration of the body fluid sample.
Acidic or alkaline dilute solution containing components to be converted
At least after the dispensing step of the body fluid sample
Adding to the reaction vessel such that a change occurs, and the color tone change data is stored in the reaction vessel while the body fluid sample is contained in the reaction vessel.
Of a step of obtaining a state, a step of obtaining an analysis result corresponding to the mixed reaction, on the basis analyzing the results of data into the color change of the data, that has a step of determining the appropriateness of the analysis results Characteristic component analysis method of body fluid. 2. The method of claim 1, wherein the step of obtaining an analysis result comprises adding the diluted solution.
2. The bodily fluid according to claim 1, which is after the adding step.
Component analysis method.