CN105938142B - Blood detection apparatus and blood detection method - Google Patents
Blood detection apparatus and blood detection method Download PDFInfo
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Abstract
The present invention relates to a blood test apparatus and a blood test method. The present invention provides a blood testing apparatus configured to measure blood cell counts and electrolytes in a blood sample. The blood test apparatus includes: a sample placement area in which a blood sample containing an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt that is not a sodium, potassium or calcium salt of EDTA is placed; and a measurement area through which a blood cell count and an electrolyte measurement of a blood sample placed in the sample placement area are performed.
Description
Cross reference to related applications
In accordance with 35 USC 119, the present application claims priority from Japanese patent application Nos. 2015-045074 and 2015-230005, the disclosures of which are incorporated herein by reference.
All publications, patent applications, and technical standards mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent application, or technical standard was specifically and individually indicated to be incorporated by reference.
Background
Technical Field
The present invention relates to a blood test apparatus and a blood test method configured to measure blood cell count and electrolytes in a blood sample.
Background
In performing blood tests, anticoagulants are used to inhibit coagulation of a blood sample to be used as a specimen. Examples of anticoagulants include citric acid, sodium citrate, ethylenediaminetetraacetic acid (EDTA), and heparin. Generally, among these anticoagulants, different anticoagulants are used for different purposes (see, for example, patent document 1 and non-patent document 1).
For example, there are reports showing: heparin has the property of readily aggregating white blood cells and the aggregation of white blood cells increases over time. There are also reports showing: in some samples, platelet aggregation is found immediately after blood is collected (see, for example, non-patent documents 2 and 3). Therefore, it is considered that: in a blood count test using heparin, white blood cells and platelets cannot be accurately measured.
When electrolyte measurement is performed with blood detection, measurement cannot be performed in the case where the anticoagulant contains Na or K. Therefore, anticoagulants without Na and K, such as heparin, are used.
Patent document 1: japanese national phase publication (JP-A) No.2008-510578
Non-patent document 1: EDTA … Selection of Anticoagulant in Blood test from Coulter No.18Sept,1992
Non-patent document 2: shiro Miwa et al Influent of variaus antibiological assays on blood count Rinsho Byori 15(5)376-380,1967
Non-patent document 3: akemi Shimasaki et al, plant organization Dual to Mixing in blood Collection Using Heaparin, the Japanese Journal of Clinical Hematology 38(4)323-330,1997
Disclosure of Invention
Problems to be solved by the invention
In the case where different anticoagulants are used for different purposes, for example, heparin may be used for electrolyte measurements, while EDTA-2Na, 2K and 3K may be used for blood count measurements. In these cases, multiple blood collection tubes containing different anticoagulants are required for measuring the blood count and electrolytes. Therefore, blood collection becomes laborious, resulting in increased detection costs and increased burden on the patient from whom blood is collected.
Accordingly, a blood detection apparatus and a blood detection method configured to measure both a blood cell count and an electrolyte in a single blood collection tube are preferable.
One embodiment of the present invention is directed to a blood detection apparatus and a blood detection method configured to measure blood cell count and electrolytes in a blood sample.
Means for solving the problems
Specific means for achieving the above object are as follows.
<1> one embodiment of the present invention is a blood test apparatus comprising: a sample placement area in which a blood sample containing an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt that is not a sodium, potassium or calcium salt of EDTA is placed; and a measurement area through which a blood cell count and an electrolyte measurement of a blood sample placed in the sample placement area are performed.
<2> the blood detecting apparatus according to <1>, wherein the measurement area has a blood cell counter configured to measure a blood cell count in the blood sample and an electrolyte measuring instrument configured to measure an electrolyte in the blood sample.
<3> the blood test device according to <1> or <2>, wherein the measurement region measures a blood cell count and electrolytes in the blood sample, and further measures at least one selected from the group consisting of C-reactive protein (CRP), Serum Amyloid A (SAA), Tumor Necrosis Factor (TNF), interleukin-1 (IL-1), α 1-antitrypsin, α 1-antichymotrypsin, α 1-acid glycoprotein, haptoglobin (haptoglobin), ceruloplasmin (cerulosmin), troponin I (tropin I), CK-MB, myoglobin (myoglobin), and cardiac-type fatty acid binding protein (H-FABP) in the blood sample.
<4> the blood test apparatus according to any one of <1> to <3>, wherein the measurement zone measures a blood cell count and an electrolyte in the blood sample, and further measures a C-reactive protein (CRP) in the blood sample.
<5> one embodiment of the present invention is a blood test method comprising: a providing step of providing a blood sample containing an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt which is not a sodium salt, a potassium salt or a calcium salt of EDTA; and a measurement step of measuring a blood cell count and an electrolyte in the blood sample.
<6> the blood detection method according to <5>, wherein the measuring step measures a blood cell count and electrolytes in the blood sample, and further measures at least one selected from the group consisting of C-reactive protein (CRP), Serum Amyloid A (SAA), Tumor Necrosis Factor (TNF), interleukin-1 (IL-1), α 1-antitrypsin, α 1-antichymotrypsin, α 1-acid glycoprotein, haptoglobin, ceruloplasmin, troponin I, CK-MB, myoglobin, and cardiac fatty acid binding protein (H-FABP) in the blood sample.
<7> the blood detection method according to <5> or <6>, wherein the measuring step measures a blood cell count and an electrolyte in the blood sample, and further, measures a C-reactive protein (CRP) in the blood sample.
<8> one embodiment of the present invention is the blood test method according to any one of <5> to <7>, wherein the EDTA salt is at least one salt selected from the group consisting of: lithium, barium, cobalt, nickel and ammonium salts of EDTA.
<9> one embodiment of the present invention is the blood test method according to any one of <5> to <8>, wherein the EDTA salt is a lithium salt of EDTA.
<10> one embodiment of the present invention is the blood test method according to any one of <5> to <9>, wherein the EDTA salt is obtained by mixing EDTA and a metal hydroxide (excluding sodium hydroxide, potassium hydroxide or calcium hydroxide) at a mass ratio of EDTA to metal hydroxide of 1:0.1 to 1: 0.6.
<11> one embodiment of the present invention is the blood test method according to any one of <5> to <10>, wherein the concentration of EDTA salt in the blood sample is 0.5mg/mL to 3.0 mg/mL.
<12> one embodiment of the present invention is the blood test method according to any one of <5> to <11>, wherein the concentration of EDTA salt in the blood sample is 0.75mg/mL to 1.5 mg/mL.
ADVANTAGEOUS EFFECTS OF INVENTION
According to one embodiment of the present invention, a blood detection apparatus and a blood detection method configured to measure a blood cell count and an electrolyte in a blood sample can be provided.
Drawings
FIGS. 1A-1C are graphs comparing the white blood cell count results between blood collection tubes 1-3(EDTA-4H + LiOH) and blood collection tube 4 (EDTA-2K). FIG. 1D is a graph comparing the white blood cell count results between blood collection tube 5 (heparin Li) and blood collection tube 4 (EDTA-2K).
FIGS. 2A-2C are graphs comparing red blood cell counts between blood collection tubes 1-3(EDTA-4H + LiOH) and blood collection tube 4 (EDTA-2K). FIG. 2D is a graph comparing the results of red blood cell counts between blood collection tube 5 (heparin Li) and blood collection tube 4 (EDTA-2K).
FIGS. 3A to 3C are graphs comparing the results of platelet count between blood collection tubes 1-3(EDTA-4H + LiOH) and blood collection tube 4 (EDTA-2K). FIG. 3D is a graph comparing the results of platelet counts between blood collection tube 5 (heparin Li) and blood collection tube 4 (EDTA-2K).
FIGS. 4A to 4C are graphs comparing CRP measurement results between the blood collection tube 1-3(EDTA-4H + LiOH) and the blood collection tube 5 (heparin Li). FIG. 4D is a graph comparing CRP measurements between blood collection tube 4(EDTA-2K) and blood collection tube 5 (heparin Li).
FIGS. 5A to 5C are graphs comparing the results of Na concentration measurement between the blood collection tube 1-3(EDTA-4H + LiOH) and the blood collection tube 5 (heparin Li). FIG. 5D is a graph comparing the results of Na concentration measurement between the blood collection tube 4(EDTA-2K) and the blood collection tube 5 (heparin Li).
FIGS. 6A to 6C are graphs comparing the results of K concentration measurements between the blood collection tube 1-3(EDTA-4H + LiOH) and the blood collection tube 5 (heparin Li). FIG. 6D is a graph comparing the results of K concentration measurements between blood collection tube 4(EDTA-2K) and blood collection tube 5 (heparin Li).
FIGS. 7A to 7C are graphs comparing the results of Cl concentration measurements between blood collection tubes 1-3(EDTA-4H + LiOH) and blood collection tube 5 (heparin Li). FIG. 7D is a graph comparing the results of Cl concentration measurements between blood collection tube 4(EDTA-2K) and blood collection tube 5 (heparin Li).
Detailed Description
Detailed Description
One embodiment of the blood test apparatus and the blood test method of the present invention is described below.
In the present specification, each numerical range represented by "… … to … … (… … to … …)" means that the range includes values described before and after "to (-)" as a lower limit and an upper limit, respectively.
[ blood testing apparatus ]
One embodiment of the blood testing apparatus of the present invention comprises a sample placement area in which a blood sample containing an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt other than the sodium, potassium or calcium salt of EDTA is placed; and a measurement area through which a blood cell count and an electrolyte measurement of the blood sample placed in the sample placement area are performed.
Traditionally, different anticoagulants have been used for different purposes. Heparin has been used for electrolyte measurements and EDTA-2Na, 2K and 3K have been used for blood count measurements. Therefore, in the case where the blood cell count and the electrolyte are to be measured, a plurality of blood collection tubes (blood sample containers) containing different anticoagulants are necessary. Therefore, blood collection becomes laborious, resulting in increased detection costs and increased burden on the patient from whom blood is collected.
In the blood test apparatus of the present embodiment, a blood sample containing an anticoagulant containing an EDTA salt other than a sodium salt, a potassium salt, or a calcium salt of EDTA is placed in a sample placement area; the blood cell count and electrolytes of the blood sample placed in the sample placement area are measured by the measurement area. Thus, without the need to use different anticoagulants for different purposes, one anticoagulant or a single blood collection tube may be used to perform the blood cell count and electrolyte measurements in the blood sample.
The blood sample to be measured using the blood test apparatus of the present embodiment is the same as the blood sample to be used in the blood test method of the present embodiment described later.
The blood test apparatus of the present embodiment includes a sample placement section in which a blood sample containing an anticoagulant containing an EDTA salt other than a sodium salt, a potassium salt, or a calcium salt of EDTA is placed. The sample-placing section is not limited as long as the sample-placing section has a configuration in which a blood sample containing an anticoagulant containing an EDTA salt can be placed, and the blood cell count and the electrolyte in the blood sample can be measured by a measurement section described later. Examples of the sample-placing section include a place where a blood collection tube (blood sample container) containing a blood sample containing an anticoagulant containing EDTA salt can be placed; and a container into which a blood sample can be injected.
The blood test apparatus of the present embodiment includes a measurement area through which a blood cell count and an electrolyte measurement of a blood sample placed in a sample placement area are performed. The measurement region may contain, for example, a nozzle (nozzle) for sucking a blood sample, and a measuring instrument to which the blood sample sucked through the nozzle is supplied to measure items.
In the measuring instrument, a device capable of measuring blood cell count (a blood cell counter) and a device capable of measuring electrolyte (an electrolyte measuring device) may be present independently, and a combination of these devices may constitute a system. Alternatively, the measurement instrument may be a single measurement device capable of measuring both the blood cell count and the electrolyte.
In the case where the blood cell count and the electrolytes in a blood sample are to be measured, these items may be measured simultaneously, or may be measured sequentially.
The blood test apparatus of the present embodiment may be configured to measure biochemical test items described later, in addition to the blood cell count and the electrolyte. For example, in the measuring instrument, a device capable of measuring a blood cell count (a hemocytometer), a device capable of measuring an electrolyte (an electrolyte measuring device), and a device capable of measuring C-reactive protein (CRP) (a CRP measuring device) may be separately present, and a combination of these devices may constitute a system. Alternatively, the measurement instrument may be a single measurement device capable of measuring the blood count, electrolytes, and CRP.
[ blood detection method ]
One embodiment of the blood detection method of the present invention includes: a providing step of providing a blood sample containing an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt which is not a sodium salt, a potassium salt or a calcium salt of EDTA; and a measurement step of measuring a blood cell count and an electrolyte in the blood sample.
The blood detection method according to the present embodiment may be performed using the blood detection apparatus described above.
With the blood detection method of the present embodiment, a single blood collection tube can be used to measure the blood count and the electrolyte. Thus, blood collection labor and testing costs may be reduced relative to the case where multiple blood collection tubes containing different anticoagulants are used to measure blood counts and electrolytes. Further, the burden on the patient from which blood is collected can be reduced.
One embodiment of the blood testing apparatus and blood testing method of the present invention uses an anticoagulant containing an ethylenediaminetetraacetic acid (EDTA) salt that is not a sodium, potassium, or calcium salt of EDTA.
EDTA inhibits blood coagulation by chelating calcium ions to form soluble complexes. Because EDTA salt is readily soluble in a solvent such as water, the anticoagulant to be used in the blood testing method may be a solution in which EDTA salt is dissolved in a solvent such as water. The anticoagulant to be used in the blood detection method may also be a solid obtained by: an EDTA salt is dissolved in a solvent such as water to prepare a solution, and then the prepared solution is dried.
The EDTA salt contained in the anticoagulant is not limited as long as the EDTA salt is a sodium salt, a potassium salt, or a calcium salt of EDTA. For example, the EDTA salt is preferably at least one salt selected from the following: lithium, barium, cobalt, nickel and ammonium salts of EDTA. In particular, the EDTA salt contained in the anticoagulant is preferably a lithium salt of EDTA.
The EDTA salt contained in the anticoagulant is preferably a metal salt obtained by mixing EDTA with a metal hydroxide (excluding sodium hydroxide, potassium hydroxide, or calcium hydroxide). The EDTA salt is more preferably a metal salt obtained by mixing EDTA with at least one metal hydroxide (more preferably, lithium hydroxide) selected from the group consisting of lithium hydroxide, barium hydroxide, cobalt hydroxide and nickel hydroxide.
The EDTA salt contained in the anticoagulant is preferably obtained by mixing EDTA and a metal hydroxide (excluding sodium hydroxide, potassium hydroxide, or calcium hydroxide) in a mass ratio of EDTA to metal hydroxide of 1:0.1 to 1: 0.6. The mass ratio of EDTA to metal hydroxide is more preferably 1:0.2 to 1:0.6, still more preferably 1:0.3 to 1: 0.5.
In one embodiment of the blood testing method of the present invention, a blood sample containing an anticoagulant is provided, and the anticoagulant provided is used to measure blood counts and electrolytes.
The concentration of EDTA salt in the blood sample to be used for the measurement of blood counts and electrolytes is preferably between 0.5mg/mL and 3.0mg/mL, more preferably between 0.75mg/mL and 1.5mg/mL, still more preferably between 0.75mg/mL and 1.25 mg/mL.
In the case where the concentration of EDTA salt is not less than 0.5mg/mL, blood coagulation can be favorably inhibited. In the case where the concentration of EDTA salt is not higher than 3.0mg/mL, hemolysis can be advantageously inhibited.
In the case where the concentration of EDTA salt is not less than 0.75mg/mL, blood coagulation can be more advantageously inhibited. In the case where the concentration of the EDTA salt is not higher than 1.5mg/mL (preferably not higher than 1.25mg/mL), hemolysis can be more advantageously suppressed, and the influence of the EDTA salt on the measurement of electrolytes (e.g., Na, K, and Cl) can be reduced, enabling more accurate measurement. In particular, in the case where the EDTA salt is a lithium salt, the effect of the EDTA salt on the measurement of the electrolyte (e.g., Na, K, and Cl) may be more advantageously reduced.
From the standpoint of preventing coagulation, the International Committee for standardization in Hematology (ICSH) recommends that 1.5mg to 2.2mg of EDTA salt be contained per 1mL of blood sample (this corresponds to an EDTA salt concentration of 1.5mg/mL to 2.2 mg/mL). Paragraph [0018] of JP-A No. 2008-Buck 510578 also describes: an EDTA concentration corresponding to about 1.5mg EDTA per 1mL of blood is effective to prevent coagulation.
In one embodiment of the invention, the concentration of the salt of EDTA, preferably the lithium salt of EDTA, may be between 0.75mg/mL and 1.5mg/mL, in which case a smaller amount of EDTA, relative to usual practice, may be used to inhibit blood clotting. In addition, the effect of EDTA salt on electrolyte (e.g., Na, K, and Cl) measurement can be reduced, enabling more accurate measurement.
The blood count to be measured by one embodiment of the blood test apparatus and the blood test method of the present invention provides information on the cellular components of blood. For example, White Blood Cells (WBC), Red Blood Cells (RBC), Hematocrit (HCT), hemoglobin level (HGB), Platelets (PLT), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), red blood cell distribution width (RDW), Platelet Distribution Width (PDW), Mean Platelet Volume (MPV), neutrophil count (NEUT), lymphocyte count (LYMPH), monocyte count (MONO), eosinophil count (EO), basophil count (BASO), reticulocyte count (RET), and reticulocyte ratio (RET%) are measured by measuring the hematocrit.
By measuring the blood count, preferably at least one of White Blood Cells (WBC), Red Blood Cells (RBC) or Platelets (PLT) is measured. More preferably, White Blood Cells (WBCs), Red Blood Cells (RBCs) and Platelets (PLTs) are measured.
Examples of items that can be measured by one embodiment of the blood test apparatus and the blood test method of the present invention include electrolytes such as sodium (Na), potassium (K), and chlorine (Cl) in addition to the above-described blood cell count.
Biochemical test items other than the blood cell count and the electrolyte can be measured (tested) by one embodiment of the blood test apparatus and the blood test method of the present invention. For example, so-called blood components can be detected qualitatively or quantitatively by biochemical means in addition to blood cell counts and electrolytes. Examples of biochemical methods include nitrogen content detection, metal detection, plasma protein-related detection, colloidal reaction (colloidal reaction), bio-pigment detection, enzyme-related detection, lipid detection, and carbohydrate metabolism detection.
In one embodiment of the blood test apparatus and the blood test method of the present invention, at least one item selected from the group consisting of, for example, C-reactive protein (CRP), Serum Amyloid A (SAA), Tumor Necrosis Factor (TNF), interleukin-1 (IL-1), α 1-antitrypsin, α 1-antichymotrypsin, α 1-acid glycoprotein, haptoglobin, ceruloplasmin, troponin I, CK-MB, myoglobin, and cardiac fatty acid binding protein (H-FABP) may be additionally measured in the blood sample.
In one embodiment of the blood test apparatus and the blood test method of the present invention, C-reactive protein (CRP) in a blood sample is preferably measured in addition to the blood cell count and the electrolyte in the blood sample. By this measurement, all of the blood count, CRP, and electrolytes in a blood sample can be measured using one anticoagulant or a single blood collection tube.
Examples of other items that can be measured in the assay include general biochemical assay items such as Creatine Phosphokinase (CPK), Blood Urea Nitrogen (BUN), Lactate Dehydrogenase (LDH), alkaline phosphatase (ALP), GOT (glutamic-oxaloacetic transaminase), GPT (glutamic-pyruvic transaminase), Triglyceride (TG), Urea (UA), Total Cholesterol (TCHO), High Density Lipoprotein Cholesterol (HDLC), Total Protein (TP), Albumin (ALB), Amylase (AMYL), gamma-glutamine transpeptidase (GGT), Creatinine (CREA), and Total Bilirubin (TBIL). One or more of these items may be measured.
Examples
An embodiment of the present invention is described below by way of examples. The invention is not limited to the embodiments.
< experiment 1-Observation of anticoagulation Property >
The anticoagulation property of the anticoagulant to be used in one embodiment of the blood detection method of the present invention is observed as follows.
[ preparation of aqueous solutions 1-3]
3 kinds of aqueous solutions (anticoagulants) containing EDTA-4H (manufactured by Dojindo Laboratories) and LiOH (manufactured by Wako pure chemical Industries, Ltd.) were prepared as follows.
< aqueous solution 1> 30mg of EDTA-4H and 11.9mg of LiOH were dissolved in 1mL of distilled water to prepare an aqueous solution 1.
< aqueous solution 2> 20mg of EDTA-4H and 7.9mg of LiOH were dissolved in 1mL of distilled water to prepare aqueous solution 2.
< aqueous solution 3> 10mg of EDTA-4H and 4.0mg of LiOH were dissolved in 1mL of distilled water to prepare an aqueous solution 3.
[ sample ]
Whole blood collected from a subject is used as a specimen (sample).
The number of samples was 43, and the samples included 31 samples from males (males) and 12 samples from females (females).
[ Experimental method ]
In a plastic container, 50. mu.L of each of the thus prepared aqueous solutions 1 to 3 was placed, followed by drying. Subsequently, 1mL of whole blood was added to each plastic container to provide samples having EDTA concentrations (Conc) in the whole blood of 1.5mg/mL, 1.0mg/mL, and 0.5mg/mL, respectively. After adding whole blood to each container, each sample was allowed to stand at room temperature for 1.5, 3.0 or 4.5 hours (90, 180 or 270 minutes), followed by observing whether coagulation and hemolysis occurred. More specifically, at 1.5, 3.0 and 4.5 hours, the coagulation state was visually observed. In addition, at 1.5, 3.0 and 4.5 hours, the specimen was centrifuged, and the hemolytic state was visually observed after centrifugation.
The results are shown in table 1.
[ Table 1]
As shown in Table 1, most samples showed no coagulation or no hemolysis at EDTA concentrations of 0.5mg/mL or 1.5mg/mL in whole blood. At a concentration of EDTA in whole blood of 1.0mg/mL, no sample showed clotting or hemolysis.
< experiment 2-Effect on measurement values of Na, K and Cl >
The effect of the anticoagulant to be used in one embodiment of the blood detection method of the present invention on the measurement values of Na, K and Cl was investigated.
[ preparation of aqueous solutions 4-11]
2 kinds of aqueous solutions (anticoagulants) containing EDTA-4H (manufactured by Dojindo Laboratories) and LiOH (manufactured by Wako pure chemical Industries, Ltd.) were prepared as follows.
< aqueous solution 4-7> 60mg of EDTA-4H and 23.7mg of LiOH were dissolved in 1mL of distilled water to prepare an aqueous solution 4. The aqueous solution 4 was diluted at a ratio of 2/3, 1/2, or 1/3 to prepare an aqueous solution. Thus providing 4 aqueous solutions 4-7 having different concentrations.
<Aqueous solution 8-11>48mg of EDTA-2NH4Dissolved in 1mL of distilled water to prepare an aqueous solution 8. The aqueous solution 8 was diluted at a ratio of 2/3, 1/2, or 1/3 to prepare an aqueous solution. Thereby providing 4 aqueous solutions 8-11 having different concentrations.
[ sample ]
Commercially available control sera were used as samples.
To 0.57mL of commercially available control serum was added 0.03mL of each of the aqueous solutions 4-11 to prepare a measurement sample. In addition, distilled water was added to a commercially available control serum to prepare a comparative measurement sample.
[ Experimental method ]
Using thus prepared measurement samples, each of Na, K and ClThe concentration was measured 5 times and significance detection was performed (significance level, 5%; significance was set to t)>2.2622). The measured value was compared with a value obtained using a comparative measurement sample to which distilled water was added.
The results are shown in tables 2-4. Table 2 corresponds to the Na concentration; table 3 corresponds to the concentration of K; table 4 corresponds to Cl concentration.
[ Table 2]
[ Table 3]
[ Table 4]
From tables 2-4, it can be inferred that: the Na, K and Cl measurements were minimally affected by the use of aqueous solution 7, aqueous solution 7 containing LiOH and 1.0mg/mL EDTA-4H.
< experiment 3-measurement of blood cell count, CRP and electrolyte >
The blood cell count (white blood cell count, red blood cell count and platelet count), CRP and electrolytes in a blood sample (whole blood) are measured by one embodiment of the blood detection method of the present invention.
[ provision of blood collecting tube 1-3]
Blood collection tubes 1-3, which contained EDTA-4H (manufactured by Dojindo Laboratories) and LiOH (manufactured by Wako Pure Chemical Industries, Ltd.) were provided as follows.
< blood collection tube 1> 300mg of EDTA-4H and 118.5mg of LiOH were dissolved in distilled water to prepare 5mL of an aqueous solution. 10.4. mu.L of the prepared aqueous solution was put into a blood collection tube, followed by drying.
< blood collection tube 2> 300mg of EDTA-4H and 118.5mg of LiOH were dissolved in distilled water to prepare 5mL of an aqueous solution. The blood collection tube was put with 8.3. mu.L of the prepared aqueous solution, followed by drying.
< blood collection tube 3> 300mg of EDTA-4H and 118.5mg of LiOH were dissolved in distilled water to prepare 5mL of an aqueous solution. 6.3. mu.L of the prepared aqueous solution was put into a blood collection tube, followed by drying.
[ Experimental method ]
To each of the above blood collection tubes 1-3, 0.5mL of whole blood (as a sample) was added to provide samples having EDTA concentrations in the whole blood of 1.25mg/mL, 1.0mg/mL, and 0.75mg/mL, respectively. Then, the blood collection tube 1-3 to which whole blood was added was used to measure the blood count, CRP, and electrolytes. The blood count, CRP, and electrolytes were measured using SB-1420 manufactured by Arkray, inc. and D-Concept D-01 and D-02 manufactured by Arkray.
[ sample for reference ]
For comparison, a blood collection tube 4 containing EDTA-2K (product name, NIHON) and a blood collection tube 5 containing heparin Li (prepared by dissolving 300mg of heparin Li in distilled water to prepare 5mL of an aqueous solution, putting 8.3. mu.L of the prepared aqueous solution into the blood collection tube, and then drying the solution) were provided. The blood counts were measured by the blood collection tube 4, and CRP and electrolytes were measured using the blood collection tube 5.
(blood cell count)
Fig. 1A to 3D show the measurement results of the blood cell count (white blood cell count, red blood cell count, and platelet count). FIGS. 1A to 1C, FIGS. 2A to 2C, and FIGS. 3A to 3C show the comparison of the blood cell count results between the blood collection tube 1-3(EDTA-4H + LiOH) and the blood collection tube 4(EDTA-2K), and FIGS. 1D, 2D, and 3D show the comparison of the blood cell count results between the blood collection tube 5 (heparin Li) and the blood collection tube 4 (EDTA-2K).
Fig. 1A to 1D show comparison of measurement results of White Blood Cell (WBC) count; fig. 2A to 2D show comparison of the measurement results of Red Blood Cell (RBC) count; fig. 3 (a) -3 (D) show a comparison of the measurement results of Platelet (PLT) counts.
As shown in fig. 1A-1C, wherein the White Blood Cell (WBC) count in each blood collection tube 1-3 is along the ordinate, the White Blood Cell (WBC) count in the blood collection tube 4 is along the abscissa, the slope of the regression equation is about 1 (satisfying the range of 0.9-1.1), and the correlation coefficient (R) is2) Not less than 0.9.
As shown in fig. 2A-2C, wherein the Red Blood Cell (RBC) count in each blood collection tube 1-3 is along the ordinate, the Red Blood Cell (RBC) count in the blood collection tube 4 is along the abscissa, the slope of the regression equation is in the range of 0.9-1.0, and the correlation coefficient (R;) is2) Not less than 0.9.
As shown in fig. 3A-3C, where the Platelet (PLT) count in each blood collection tube 1-3 is along the ordinate, the Platelet (PLT) count in the blood collection tube 4 is along the abscissa, the slope of the regression equation is in the range of 0.9-1.0, and the correlation coefficient (R;) is2) Not less than 0.9.
Therefore, by using a blood collection tube containing EDTA-4H + LiOH, the performance of the blood cell count (white blood cell count, red blood cell count, and platelet count) measurement can be ensured.
As shown, by using EDTA-4H + LiOH containing blood collection tubes, the blood cell count (white blood cell count, red blood cell count, and platelet count) can be measured more accurately than with heparin Li containing blood collection tubes (fig. 1D, 2D, 3D).
(CRP)
Fig. 4A to 4D show the measurement results of CRP (C-reactive protein). FIGS. 4 (A) -4(C) show a comparison of CRP measurements between the blood collection tubes 1-3(EDTA-4H + LiOH) and the blood collection tube 5 (heparin Li), and FIG. 4D shows a comparison of CRP measurements between the blood collection tube 4(EDTA-2K) and the blood collection tube 5 (heparin Li).
As shown in fig. 4A-4C, where the CRP measurement in each blood collection tube 1-3 is along the ordinate, blood collectionCRP measurements in header 5 along the abscissa, the slope of the regression equation is 0.9-1.0, the correlation coefficient (R)2) Not less than 0.9.
Therefore, by using the blood collection tube containing EDTA-4H + LiOH, the performance of CRP measurement can be ensured.
(electrolyte)
Fig. 5A to 7D show the measurement results of electrolytes (Na, K, and Cl). In each figure, FIGS. 5A to 5C, FIGS. 6A to 6C, and FIGS. 7A to 7C show the comparison of the electrolyte measurement results between the blood collection tube 1-3(EDTA-4H + LiOH) and the blood collection tube 5 (heparin Li), and FIGS. 5D, 6D, and 7D show the comparison of the electrolyte measurement results between the blood collection tube 4(EDTA-2K) and the blood collection tube 5 (heparin Li).
As shown in fig. 6D, where the blood collection tube 4(EDTA-2K) was used, potassium (K) could not be measured (due to out of range).
In fig. 7A to 7D, the measured value of chlorine (Cl) obtained using the blood collection tube 5 (heparin Li) is provided as a control, and judgment is made based on the difference (% deviation) from the control.
As shown in FIGS. 5A-5C, where the measured concentration of sodium (Na) in each blood collection tube 1-3 is along the ordinate, the measured concentration of sodium (Na) in blood collection tube 5 is along the abscissa, the slope of the regression equation is not less than 0.8, and the correlation coefficient (R) is2) Not less than 0.9.
On the other hand, as shown in FIG. 5D, where the measured concentration of sodium (Na) in the blood collection tube 4 is along the ordinate and the measured concentration of sodium (Na) in the blood collection tube 5 is along the abscissa, the slope of the regression equation is about 0.58, the correlation coefficient (R)2) Less than 0.9.
Thus, as shown in FIGS. 5A to 5D, by using the EDTA-4H + LiOH-containing blood collection tube, the sodium (Na) concentration can be measured more accurately than in the case of using the EDTA-2K-containing blood collection tube (FIG. 5D).
As shown in fig. 6A to 6D, in the case of using the blood collection tube containing EDTA-2K, the potassium (K) concentration could not be measured (fig. 6D). However, in the case of using a blood collection tube containing EDTA-4H + LiOH, the potassium (K) concentration can be measured.
As shown in FIGS. 7A to 7D, the difference in value between the case of using the EDTA-4H + LiOH-containing blood collection tube and the case of using the heparin Li-containing blood collection tube was mostly in the range of-10 to + 10%. However, the difference in value between the case of using the EDTA-2K-containing blood collection tube (FIG. 7D) and the case of using the heparin Li-containing blood collection tube was large and some data exceeded + 15%.
Thus, as shown in FIGS. 7A to 7D, by using the EDTA-4H + LiOH-containing blood collection tube, the chlorine (Cl) concentration can be measured more accurately than in the case of using the EDTA-2K-containing blood collection tube (FIG. 7D).
Thus, by one embodiment of the blood detection method of the present invention, it is possible to measure the blood cell count and the electrolytes in a blood sample (whole blood), and also to measure CRP in a blood sample. Thus, by the blood detection method, the blood cell count (white blood cell count, red blood cell count, and platelet count), CRP, and electrolytes can be measured in a single blood collection tube, so that the number of blood collection tubes can be reduced. Therefore, the labor and cost of blood collection can be reduced, and the burden on the patient can be reduced.
Claims (7)
1. A method of blood testing, comprising:
providing a blood sample containing an anticoagulant comprising an ethylenediaminetetraacetic acid salt that is not a sodium, potassium or calcium salt of EDTA; and
measuring the blood cell count and electrolytes in said blood sample, wherein said EDTA salt is at least one salt selected from the group consisting of: lithium, barium, cobalt, nickel and ammonium salts of EDTA.
2. The blood detection method of claim 1, further comprising measuring in the blood sample at least one item selected from the group consisting of C-reactive protein (CRP), Serum Amyloid A (SAA), Tumor Necrosis Factor (TNF), interleukin-1 (IL-1), α 1-antitrypsin, α 1-antichymotrypsin, α 1-acid glycoprotein, haptoglobin, ceruloplasmin, troponin I, CK-MB, myoglobin, and cardiac fatty acid binding protein (H-FABP).
3. The blood detection method according to claim 1 or claim 2, wherein C-reactive protein (CRP) in the blood sample is measured.
4. The blood detection method of claim 1 or claim 2, wherein the EDTA salt is a lithium salt of EDTA.
5. The blood detection method according to claim 1 or claim 2, wherein the EDTA salt is obtained by mixing EDTA and a metal hydroxide excluding sodium hydroxide, potassium hydroxide or calcium hydroxide in a mass ratio of EDTA to metal hydroxide of 1:0.1 to 1: 0.6.
6. The method according to claim 1 or claim 2, wherein the concentration of EDTA salt in the blood sample is 0.5mg/mL to 3.0 mg/mL.
7. The method according to claim 1 or claim 2, wherein the concentration of EDTA salt in the blood sample is 0.75mg/mL to 1.5 mg/mL.
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