JP2000180439A - Method for pretreating whole blood for measuring saccharified hemoglobin - Google Patents
Method for pretreating whole blood for measuring saccharified hemoglobinInfo
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- JP2000180439A JP2000180439A JP10362559A JP36255998A JP2000180439A JP 2000180439 A JP2000180439 A JP 2000180439A JP 10362559 A JP10362559 A JP 10362559A JP 36255998 A JP36255998 A JP 36255998A JP 2000180439 A JP2000180439 A JP 2000180439A
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- albumin
- whole blood
- glycated hemoglobin
- complex
- hemoglobin
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、酵素法による、全
血中の糖化ヘモグロビンの測定方法、並びに糖化ヘモグ
ロビン測定用全血の前処理方法及び該全血の前処理試薬
に関する。TECHNICAL FIELD The present invention relates to a method for measuring glycated hemoglobin in whole blood by an enzymatic method, a method for pretreating whole blood for measuring glycated hemoglobin, and a reagent for pretreating the whole blood.
【０００２】[0002]
【従来技術】糖化ヘモグロビン（HbA1）とは、拡散によ
り赤血球に入り込んで、血液中の糖（単糖類）の還元末
端と、ヘモグロビンのα鎖およびβ鎖の遊離のアミノ基
が非酵素的にシッフ塩基により結合して形成された不安
定なアルドイミン（不安定型）が、さらにアマドリ転移
により安定なケトアミン（安定型）へと移行することに
より生成されるアマドリ化合物の一種である。糖化ヘモ
グロビンの濃度は過去１〜２ヶ月間の血液中の平均的な
糖濃度を反映すると言われていることから、糖尿病の診
断マーカーとして広く利用されている。2. Description of the Related Art Glycated hemoglobin (HbA 1 ) enters erythrocytes by diffusion, and the reducing end of sugar (monosaccharide) in blood and the free amino groups of α and β chains of hemoglobin are non-enzymatically. It is a kind of Amadori compound generated by transferring an unstable aldoimine (unstable type) formed by binding with a Schiff base to a stable ketoamine (stable type) by Amadori rearrangement. Since it is said that the concentration of glycated hemoglobin reflects the average sugar concentration in blood for the past one to two months, it is widely used as a diagnostic marker for diabetes.
【０００３】従来の糖化ヘモグロビンの測定法として
は、高速液体クロマトグラフィー（HPLC）法〔Chromato
gr.Sci.10:659(1979)〕、アフィニティーカラム法〔Cli
n.Chem.28:2088(1982)〕、電気泳動法〔Clin.Chem.26:1
598(1980)〕、チオバルビツール酸法〔Clin.Chim.Acta
112:179-204(1981)〕、免疫学的測定法〔JJCLA 18:620
(1983), 機器・試薬16:33-37(1993)〕等がある。As a conventional method for measuring glycated hemoglobin, a high performance liquid chromatography (HPLC) method [Chromato
gr.Sci. 10: 659 (1979)), affinity column method (Cli
n.Chem. 28: 2088 (1982)), electrophoresis (Clin.Chem. 26: 1)
598 (1980)), thiobarbituric acid method (Clin.
112: 179-204 (1981)), immunoassay (JJCLA 18: 620
(1983), Instrument / Reagent 16: 33-37 (1993)].
【０００４】しかしながら、これらの方法は何れも何ら
かの問題点を有している。先ず、ＨＰＬＣ法は、最も普
及しており、測定精度や再現性が良いが、高価で特別な
機器を必要としたり、単位時間当たりの処理検体数が少
ないという点で問題がある。[0004] However, each of these methods has some problems. First, the HPLC method is the most widespread and has good measurement accuracy and reproducibility, but has problems in that expensive and special equipment is required and the number of processed samples per unit time is small.
【０００５】アフィニティーカラム法、電気泳動法、チ
オバルビツール酸法は何れの方法も操作性、処理能力の
点で問題がある。All of the affinity column method, electrophoresis method and thiobarbituric acid method have problems in operability and processing ability.
【０００６】免疫学的測定法は、近年開発された方法で
自動分析装置での測定が可能なため、操作が簡便で処理
検体数も多く、ＨＰＬＣ法との相関のよい方法ではある
が、ＨＰＬＣ法に比べ再現性の点で問題がある。The immunoassay is a method developed in recent years, which can be measured by an automatic analyzer, so that the operation is simple, the number of processed samples is large, and the method has a good correlation with the HPLC method. There is a problem in reproducibility compared with the method.
【０００７】近年、上記した如き問題を解決するため
に、糖化ヘモグロビン等のアマドリ化合物を基質として
過酸化水素を生成する酸化還元酵素を利用した糖化ヘモ
グロビン測定法が提案されている（特開平2-195899号公
報、特開平2-195900号公報、特開平3-155780号公報、特
開平4-4874号公報、特開平5-192193号公報、特開平6-46
846号公報、特開平7-289253号公報、特開平8-154672号
公報、特開平8-336386号公報、特開平10-201473号公
報）。[0007] In recent years, in order to solve the above-mentioned problems, a method of measuring glycated hemoglobin using an oxidoreductase that generates hydrogen peroxide using an Amadori compound such as glycated hemoglobin as a substrate has been proposed (Japanese Patent Laid-Open No. Hei 2- 195899, JP-A-2-195900, JP-A-3-155780, JP-A-4-4874, JP-A-5-192193, JP-A-6-46
846, JP-A-7-289253, JP-A-8-154672, JP-A-8-336386, JP-A-10-201473).
【０００８】しかしながら、血液中には、アマドリ化合
物として糖化ヘモグロビン以外に糖化アルブミンも含ま
れており、アマドリ化合物を基質とする酸化還元酵素を
使用し、全血を試料として糖化ヘモグロビンの測定を行
った場合、糖化アルブミンによる正誤差が生じる。この
ような正誤差を回避するためには、全血から血漿成分を
取り除くための操作、例えば遠心分離処理等の煩雑な操
作が必要であった。However, glycated albumin is also contained in blood in addition to glycated hemoglobin as an Amadori compound, and glycated hemoglobin was measured using whole blood as a sample using an oxidoreductase using the Amadori compound as a substrate. In this case, saccharified albumin causes a correct error. In order to avoid such a correct error, an operation for removing plasma components from whole blood, for example, a complicated operation such as a centrifugal separation process was required.
【０００９】そのため、上記した如き問題を解決した、
酵素法による糖化ヘモグロビンの測定を行うための、効
率的な全血の前処理方法が強く望まれているが、未だ実
現されていない。Therefore, the above-described problem has been solved.
Although an efficient pretreatment method of whole blood for measuring glycated hemoglobin by an enzymatic method is strongly desired, it has not been realized yet.
【００１０】[0010]
【発明が解決しようとする課題】上記した如き状況に鑑
み、本発明が解決しようとする課題は、酵素反応を利用
して全血中の糖化ヘモグロビンを測定する際に生じる糖
化アルブミンの影響を回避し得る、糖化ヘモグロビン測
定方法、並びに糖化ヘモグロビン測定用全血の前処理方
法および前処理用試薬を提供することにある。SUMMARY OF THE INVENTION In view of the above situation, an object of the present invention is to avoid the effects of glycated albumin generated when measuring glycated hemoglobin in whole blood using an enzymatic reaction. It is an object of the present invention to provide a method for measuring glycated hemoglobin, a method for pretreating whole blood for measuring glycated hemoglobin, and a reagent for pretreatment.
【００１１】[0011]
【課題を解決するための手段】本発明は上記課題を解決
する目的でなされたものであり、酵素法により全血中の
糖化ヘモグロビンを測定する方法に於て、全血をアルブ
ミンとコンプレックスを形成する物質で処理し、次いで
当該測定に付することを特徴とする、血液試料中の糖化
ヘモグロビンの測定方法の発明である。SUMMARY OF THE INVENTION The present invention has been made for the purpose of solving the above-mentioned problems. In a method for measuring glycated hemoglobin in whole blood by an enzymatic method, the whole blood is formed into a complex with albumin. The present invention relates to a method for measuring glycated hemoglobin in a blood sample, wherein the method is performed by treating with a substance to be treated and then subjecting to the measurement.
【００１２】また、全血をアルブミンとコンプレックス
を形成する物質で処理することを特徴とする、糖化ヘモ
グロビン測定用全血の前処理方法の発明である。[0012] The present invention also provides a method for pretreating whole blood for measuring glycated hemoglobin, which comprises treating whole blood with a substance which forms a complex with albumin.
【００１３】更に、アルブミンとコンプレックスを形成
する物質を含有してなる糖化ヘモグロビン測定用全血の
前処理試薬の発明である。[0013] The present invention further provides a reagent for pretreating whole blood for measuring glycated hemoglobin, which contains a substance forming a complex with albumin.
【００１４】本発明者らは、全血を試料とした酵素法に
よる糖化ヘモグロビンの測定であって全血中の糖化アル
ブミンによる測定値への悪影響を回避し得る高精度な糖
化ヘモグロビンの測定法を開発すべく鋭意検討を行った
結果、全血を、アルブミンとコンプレックスを作る物質
で前処理した後に測定に供すれば、糖化アルブミンの影
響を受けることなく、全血中の糖化ヘモグロビンを高精
度に測定し得ることを見いだし、本発明を完成させるに
至った。The present inventors have developed a highly accurate method for measuring glycated hemoglobin using whole blood as a sample by an enzymatic method, which method can avoid the adverse effect of glycated albumin in whole blood on measured values. As a result of intensive studies to develop, if the whole blood is pre-treated with a substance that forms a complex with albumin and then subjected to measurement, the glycated hemoglobin in whole blood can be accurately measured without being affected by glycated albumin. They have found that they can be measured and have completed the present invention.
【００１５】本発明に係るアルブミンとコンプレックス
を形成する物質としては、例えば抗アルブミン抗体やブ
ルー色素Cibacron BlueF3G-Aなどが挙げられ、中でも抗
アルブミン抗体が好ましいものとして挙げられる。The substance forming a complex with albumin according to the present invention includes, for example, an anti-albumin antibody and a blue dye Cibacron BlueF3G-A, and among them, an anti-albumin antibody is preferable.
【００１６】また、ブルー色素Cibacron BlueF3G-Aは、
適当な担体に共有結合で固定化したもの（ブルートリア
クリル：和光純薬工業（株）商品名、ハイトラップブル
ー：ファルマシア商品名）でもよい。The blue dye Cibacron BlueF3G-A is
What was immobilized on a suitable carrier by a covalent bond (blue triacryl: trade name of Wako Pure Chemical Industries, Ltd., high trap blue: trade name of Pharmacia) may be used.
【００１７】尚、抗アルブミン抗体は、市販品でも常法
により適宜調製されたものでもよく、また、モノクロー
ナル抗体でもポリクローナル抗体でもよいが、ポリクロ
ーナル抗体がより好ましい。The anti-albumin antibody may be a commercially available product or one prepared as appropriate by a conventional method. A monoclonal antibody or a polyclonal antibody may be used, but a polyclonal antibody is more preferable.
【００１８】本発明に係るアルブミンとコンプレックス
を形成する物質の使用濃度としては、全血中の糖化ヘモ
グロビンを測定する際に生じる糖化アルブミンによる影
響を抑止し得る濃度であればよく特に限定されないが、
糖化ヘモグロビン測定用全血を処理する際に、本発明に
係るアルブミンとコンプレックスを形成する物質の濃度
がアルブミン濃度に対し、通常1/50〜100倍、好ましく
は1/10〜10倍となるように添加される。The concentration of the substance that forms a complex with albumin according to the present invention is not particularly limited as long as it is a concentration that can suppress the effect of glycated albumin generated when measuring glycated hemoglobin in whole blood.
When processing glycated hemoglobin measurement whole blood, the concentration of the substance forming a complex with albumin according to the present invention is usually 1/50 to 100 times, preferably 1/1/10 to 10 times the albumin concentration. Is added to
【００１９】例えば、アルブミンとコンプレックスを形
成する物質として抗アルブミン抗体を使用する場合に
は、サンプル中のアルブミンの濃度にもよるが、サンプ
ルと前処理用試薬混合時の抗アルブミン抗体濃度が通常
0.08〜400mgAb/ml、好ましくは0.4〜40 mgAb/mlとなる
ように添加される。For example, when an anti-albumin antibody is used as a substance forming a complex with albumin, the concentration of the anti-albumin antibody when the sample and the pretreatment reagent are mixed usually depends on the concentration of albumin in the sample.
It is added so as to be 0.08 to 400 mgAb / ml, preferably 0.4 to 40 mgAb / ml.
【００２０】本発明の糖化ヘモグロビン測定用全血の前
処理方法は、本発明に係わるアルブミンとコンプレック
スを形成する物質を添加することにより行われる。The method for pretreating whole blood for measuring glycated hemoglobin of the present invention is carried out by adding a substance which forms a complex with albumin according to the present invention.
【００２１】尚、本発明の前処理方法を行って得られた
試料中の糖化ヘモグロビンの測定は例えば特開平2-1958
99号公報、特開平2-195900号公報、特開平 5-192193号
公報、特開平6-46846号公報、特開平7-289253号公報、
特開平8-154672号公報、特開平8-336386号公報などに記
載の自体公知の酵素法に準じて行えばよい。The measurement of glycated hemoglobin in a sample obtained by performing the pretreatment method of the present invention is described in, for example, Japanese Patent Application Laid-Open No. 2-1958.
No. 99, JP-A-2-195900, JP-A-5-192193, JP-A-6-46846, JP-A-7-289253,
What is necessary is just to carry out according to the per se known enzymatic method described in Unexamined-Japanese-Patent No. 8-154672, 8-336386, etc.
【００２２】以下に、実施例を挙げて本発明をさらに詳
細に説明するが、本発明はこれらにより何ら限定される
ものではない。Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.
【００２３】[0023]
【実施例】実施例１ 糖化ヘモグロビンの測定 （１）糖化ヘモグロビン量の測定 (ａ)検体及び試薬 検体：EDTA採血全血１，２及び３ 試薬：ｉ )54mg Ab/ml抗アルブミン抗体溶液 ii )ペプシン液：0.6mg/ml ペプシン in 0.01N HCl iii)カルボキシペプチダーゼ(CPase)液：3mg/ml CPaseA
及び3mg/mlCPaseY含有50mM 2-モルホリノエタンスルホ
ン酸(MES)緩衝液(pH6.0)EXAMPLES Example 1 Measurement of Glycated Hemoglobin (1) Measurement of Glycated Hemoglobin (a) Specimen and Reagent Specimen: EDTA Blood Whole Blood 1, 2 and 3 Reagent: i) 54 mg Ab / ml anti-albumin antibody solution ii) Pepsin solution: 0.6 mg / ml pepsin in 0.01N HCl iii) Carboxypeptidase (CPase) solution: 3 mg / ml CPaseA
And 3 mg / ml CPaseY-containing 50 mM 2-morpholinoethanesulfonic acid (MES) buffer (pH 6.0)
【００２４】(ｂ)糖化ヘモグロビン測定用試料の調製 検体1mlに、水2.5mlを添加し溶血させ、その後に54mg/m
l Ab/ml抗アルブミン抗体2.5mlを添加混合し全量を6ml
とし、これを糖化ヘモグロビン測定用試料とした。(B) Preparation of a sample for measuring glycated hemoglobin 2.5 ml of water was added to 1 ml of a sample to cause hemolysis, followed by 54 mg / m 2
l Add 2.5 ml of Ab / ml anti-albumin antibody and mix to make a total of 6 ml
This was used as a sample for measuring glycated hemoglobin.
【００２５】(ｃ)プロテアーゼ処理 上記(ｂ)で得られた糖化ヘモグロビン測定試料6mlにペ
プシン液を1ml添加し塩酸でpH2に調整し、全量を8mlと
した。この調製液を35℃で30分間インキュベイトするこ
とによりペプシン処理を行った。インキュベイト終了
後、反応液に更に50mM MES緩衝液(pH6.0) 1mlとCPase液
1mlを加え、水酸化ナトリウムでpH6.0に調整し、全量を
12mlとし、更に35℃で3日間インキュベイトすることに
よりCPase処理を行った。反応終了後、反応液に200mM 2
-〔4-(2-ヒドロキシエチル)-1-ピペラジニル〕エタンス
ルホン酸(HEPES)緩衝液(pH7.5)を1.5ml加え、水酸化ナ
トリウムでpH7.5に調整し、全量を15mlとし、これをプ
ロテアーゼ処理ヘモグロビン溶液とした。(C) Protease treatment 1 ml of a pepsin solution was added to 6 ml of the glycated hemoglobin measurement sample obtained in the above (b), and the pH was adjusted to 2 with hydrochloric acid to make the total amount 8 ml. This preparation was incubated at 35 ° C. for 30 minutes to perform pepsin treatment. After incubation, add 1 ml of 50 mM MES buffer (pH 6.0) and CPase solution to the reaction solution.
Add 1 ml, adjust to pH 6.0 with sodium hydroxide, and add
The volume was adjusted to 12 ml, and the mixture was incubated at 35 ° C. for 3 days to perform a CPase treatment. After the reaction is completed, add 200 mM 2
-1.5 ml of (4- (2-hydroxyethyl) -1-piperazinyl) ethanesulfonic acid (HEPES) buffer (pH 7.5) was added, and the mixture was adjusted to pH 7.5 with sodium hydroxide to make the total amount 15 ml. Was used as a protease-treated hemoglobin solution.
【００２６】(ｄ)糖化ヘモグロビン量の測定 上記(ｃ)で得られたプロテアーゼ処理ヘモグロビン溶液
中の糖化ヘモグロビン量は日立557形分光光度計（日立
製作所（株）製）を用い、下記の如くして行った。(D) Measurement of the amount of glycated hemoglobin The amount of glycated hemoglobin in the protease-treated hemoglobin solution obtained in (c) above was determined as follows using a Hitachi 557 type spectrophotometer (manufactured by Hitachi, Ltd.). I went.
【００２７】発色液：以下の試薬を含有する200mM HEPE
S緩衝液(pH7.5)を発色液とした。 N-エチル-N-(2-ヒドロキシ-3-スルホプロピル)-3,5- ジメトキシ-4-フルオロアニリン(FDAOS) 360mM 4-アミノアンチピリン(4AA) 1350mM ペルオキシダーゼ(POD) 3.6U/ml フルクトシルアミノ酸オキシダーゼ(FAOD) 9U/mlColoring solution: 200 mM HEPE containing the following reagents
S buffer (pH 7.5) was used as a color developing solution. N-ethyl-N- (2-hydroxy-3-sulfopropyl) -3,5-dimethoxy-4-fluoroaniline (FDAOS) 360 mM 4-aminoantipyrine (4AA) 1350 mM peroxidase (POD) 3.6 U / ml fructosyl amino acid Oxidase (FAOD) 9U / ml
【００２８】上記(ａ)で得られたプロテアーゼ処理ヘモ
グロビン溶液2mlに発色液1mlを加え、37℃で10分間イン
キュベイトした後、600 nmの吸光度を測定した。次に検
体として生理食塩水及び市販の糖化ヘモグロビン標準を
用いて同様の操作を行い、これを用いて検量線を作成
し、検体１，２及び３中に含まれる糖化ヘモグロビン量
(g/dl)を求めた。1 ml of a coloring solution was added to 2 ml of the protease-treated hemoglobin solution obtained in the above (a), and the mixture was incubated at 37 ° C. for 10 minutes, and the absorbance at 600 nm was measured. Next, the same operation was performed using a physiological saline solution and a commercially available glycated hemoglobin standard as a sample, and a calibration curve was prepared using the same, and the amount of glycated hemoglobin contained in the samples 1, 2, and 3 was determined.
(g / dl) was determined.
【００２９】（２）ヘモグロビン量の測定 検体：上記（１）で用いたものと同じ。 和光純薬工業（株）製のヘモグロビンＢ-テストワコー
（ＳＬＳ−ヘモグロビン法）を使用して上記検体中のヘ
モグロビン量(g/dl)を求めた。(2) Measurement of hemoglobin amount Specimen: Same as that used in (1) above. The hemoglobin amount (g / dl) in the sample was determined using hemoglobin B-Test Wako (SLS-hemoglobin method) manufactured by Wako Pure Chemical Industries, Ltd.
【００３０】（３）ヘモグロビン中の糖化ヘモグロビン
含有率（％）の算定 下記計算式から糖化ヘモグロビン含有率を算定した。結
果を表１に示す。 (3) Calculation of glycated hemoglobin content (%) in hemoglobin The glycated hemoglobin content was calculated from the following formula. Table 1 shows the results.
【００３１】比較例１ 実施例１と同じ検体を用い、実施例１（１）（ｂ）に於
て、抗アルブミン抗体2.5mlの代わりに水2.5mlを添加し
た以外は実施例１と同様にして測定を行い、検体中の糖
化ヘモグロビン含有率（％）を求めた。結果を表１に併
せて示す。Comparative Example 1 The same sample as in Example 1 was used, except that 2.5 ml of water was added instead of 2.5 ml of the anti-albumin antibody in Example 1 (1) (b). The glycated hemoglobin content (%) in the sample was determined. The results are shown in Table 1.
【００３２】参考例１ 実施例と同じ検体について、東ソー(株)の自動グリコヘ
モグロビン分析計（ＨＰＬＣ法）を用いて糖化ヘモグロ
ビン含有率(％)を求めた。結果を表１に併せて示す。Reference Example 1 The glycated hemoglobin content (%) of the same sample as in the example was determined using an automatic glycohemoglobin analyzer (HPLC method) manufactured by Tosoh Corporation. The results are shown in Table 1.
【００３３】表 １ Table 1
【００３４】表１の結果から明らかな如く、本発明の前
処理方法で検体を処理することにより、サンプル中の糖
化ヘモグロビン含有率（％）の値がＨＰＬＣ法で求めた
糖化ヘモグロビン含有率（％）の値と極めて近い値とな
ることが判る。As is clear from the results shown in Table 1, by treating the sample with the pretreatment method of the present invention, the value of the glycated hemoglobin content (%) in the sample was determined by the HPLC method. ) Is very close to the value.
【００３５】一方、検体を本発明の前処理方法で処理せ
ずに測定を行った場合の糖化ヘモグロビン含有率（％）
の値は、前処理を行った場合やＨＰＬＣ法で測定した場
合に比べて有意に高値となることが判る。言い換えれ
ば、本発明の前処理方法で検体を処理しなかった場合に
は、糖化アルブミンの影響で正誤差が生じることが判
る。On the other hand, the glycated hemoglobin content (%) when the sample was measured without being treated by the pretreatment method of the present invention.
It can be seen that the value of is significantly higher than the value obtained when the pretreatment was performed or when the value was measured by the HPLC method. In other words, it can be seen that when the sample was not processed by the pretreatment method of the present invention, a correct error occurs due to the influence of glycated albumin.
【００３６】実施例２ 検体として、EDTA採血全血検体４〜２２を用いて、実施
例１と同様にして測定を行い、糖化ヘモグロビン含有率
（％）を求めた。結果を表２に示す。Example 2 The same measurement as in Example 1 was performed using EDTA-collected whole blood samples 4 to 22 as samples, and the glycated hemoglobin content (%) was determined. Table 2 shows the results.
【００３７】参考例２ 実施例２と同じ検体について、ベーリンガーマンハイム
（株）製のリキテックHbA1cII（免疫阻害比濁法）を使
用して、ヘモグロビンA1c含有率（％）を求めた。結果
を表２に併せて示す。また実施例２の糖化ヘモグロビン
含有率（％）と参考例２のヘモグロビンA1c含有率
（％）とから得られる相関図を図１に示す。 表 ２ [0037] The same specimens as in Reference Example 2 Example 2, using the Boehringer Mannheim Co., Ltd. Rikitekku HbA 1 cII (immunoinhibition nephelometry) was determined hemoglobin A 1 c content rate (%). The results are shown in Table 2. FIG. 1 shows a correlation diagram obtained from the glycated hemoglobin content (%) of Example 2 and the hemoglobin A 1 c content (%) of Reference Example 2. Table 2
【００３８】表２及び図１から明らかな如く、本発明の
前処理方法で検体を処理することにより、検体中の糖化
ヘモグロビン含有率（％）の値は免疫阻害比濁法で求め
たヘモグロビンA1c含有率（％）の値と良好な相関を示
すことが判る。As is clear from Table 2 and FIG. 1, by treating the sample by the pretreatment method of the present invention, the value of the glycated hemoglobin content (%) in the sample was determined by the hemoglobin A determined by the immunoinhibition nephelometry. It turns out that it shows a good correlation with the value of 1 c content (%).
【００３９】尚、本発明の方法で求めた糖化ヘモグロビ
ン含有率（％）の値は、免疫阻害比濁法で求めたヘモグ
ロビンA1c含有率（％）の値に比べ若干高値になってい
るが、これは、糖化アルブミンの影響が回避されていな
いのではなく、本発明の方法ではヘモグロビンA1c以外
の糖化ヘモグロビンも測定しているためである。[0039] The value of glycated hemoglobin content was determined by the method of the present invention (%) is made slightly higher than the value of the hemoglobin A 1 c content determined by immune inhibition nephelometry (%) but this is not the influence of glycated albumin is not avoided, the method of the present invention is because it is also measured glycated hemoglobin other than hemoglobin a 1 c.
【００４０】[0040]
【発明の効果】以上述べた如く、本発明は、糖化アルブ
ミンの影響を受けることなく、酵素法で全血中の糖化ヘ
モグロビン量を高精度に測定し得る測定方法及び糖化ヘ
モグロビン測定用全血の前処理方法並びにこれに用いる
前処理試薬を提供するものであり、本発明を実施するこ
とにより、全血を血球と血漿に分離するという煩雑な操
作を行うことなく全血中の糖化ヘモグロビンを酵素法に
より測定できるという効果を奏する。As described above, the present invention provides a method for measuring the amount of glycated hemoglobin in whole blood with high accuracy by an enzyme method without being affected by glycated albumin, and a method for measuring glycated hemoglobin for whole blood. The present invention provides a pretreatment method and a pretreatment reagent used for the same.By implementing the present invention, glycated hemoglobin in whole blood can be converted to an enzyme without performing a complicated operation of separating whole blood into blood cells and plasma. This has the effect of being measurable by the method.
【図１】参考例２で得られたヘモグロビンA1c含有率
(％)の値と実施例２で得られた糖化ヘモグロビン含有率
(％)の値の相関を示す。FIG. 1 shows the hemoglobin A 1c content obtained in Reference Example 2.
(%) Value and glycated hemoglobin content obtained in Example 2
(%) Shows the correlation between the values.
フロントページの続き Ｆターム(参考） 2G045 AA01 BA01 BA13 BB39 BB50 BB51 CA25 DA38 DA45 DA48 FA29 FB01 FB03 FB06 GC10 4B063 QA01 QA19 QQ03 QQ68 QQ79 QR16 QR48 QR51 QS11 QS20 QX01 Continued on the front page F-term (reference) 2G045 AA01 BA01 BA13 BB39 BB50 BB51 CA25 DA38 DA45 DA48 FA29 FB01 FB03 FB06 GC10 4B063 QA01 QA19 QQ03 QQ68 QQ79 QR16 QR48 QR51 QR51 QS11 QS20 QX01
Claims (6)
ビンを測定する方法に於て、全血をアルブミンとコンプ
レックスを形成する物質で処理し、次いで当該測定に付
することを特徴とする、全血中の糖化ヘモグロビンの測
定方法。1. A method for measuring glycated hemoglobin in a blood sample by an enzyme method, wherein the whole blood is treated with a substance which forms a complex with albumin, and then subjected to the measurement. Method for measuring glycated hemoglobin in food.
物質が抗アルブミン抗体である請求項１に記載の測定方
法。2. The method according to claim 1, wherein the substance forming a complex with albumin is an anti-albumin antibody.
形成する物質で処理することを特徴とする、糖化ヘモグ
ロビン測定用全血の前処理方法。3. A pretreatment method for whole blood for measuring glycated hemoglobin, which comprises treating whole blood with a substance which forms a complex with albumin.
物質が抗アルブミン抗体である請求項３に記載の前処理
方法。4. The pretreatment method according to claim 3, wherein the substance forming a complex with albumin is an anti-albumin antibody.
物質を含有してなる、糖化ヘモグロビン測定用全血の前
処理試薬。5. A pretreatment reagent for whole blood for measuring glycated hemoglobin, comprising a substance forming a complex with albumin.
物質が抗アルブミン抗体である請求項５に記載の前処理
試薬。6. The pretreatment reagent according to claim 5, wherein the substance forming a complex with albumin is an anti-albumin antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10362559A JP2000180439A (en) | 1998-12-21 | 1998-12-21 | Method for pretreating whole blood for measuring saccharified hemoglobin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10362559A JP2000180439A (en) | 1998-12-21 | 1998-12-21 | Method for pretreating whole blood for measuring saccharified hemoglobin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2000180439A true JP2000180439A (en) | 2000-06-30 |
Family
ID=18477167
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10362559A Withdrawn JP2000180439A (en) | 1998-12-21 | 1998-12-21 | Method for pretreating whole blood for measuring saccharified hemoglobin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2000180439A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003033729A1 (en) * | 2001-10-11 | 2003-04-24 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| WO2005056823A1 (en) | 2003-12-12 | 2005-06-23 | Arkray, Inc. | Method of measuring saccharified amine |
| JP4834771B2 (en) * | 2006-07-21 | 2011-12-14 | エフ．ホフマン−ラ ロシュ アーゲー | Hemoglobin digestion reagent |
| USRE45074E1 (en) | 2001-10-11 | 2014-08-12 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| JP2019086450A (en) * | 2017-11-09 | 2019-06-06 | 国立大学法人弘前大学 | Analysis device for whole blood albumin, and analysis method for whole blood albumin |
-
1998
- 1998-12-21 JP JP10362559A patent/JP2000180439A/en not_active Withdrawn
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003033729A1 (en) * | 2001-10-11 | 2003-04-24 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| US7449305B2 (en) | 2001-10-11 | 2008-11-11 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| USRE43795E1 (en) | 2001-10-11 | 2012-11-06 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| USRE45074E1 (en) | 2001-10-11 | 2014-08-12 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| USRE45626E1 (en) | 2001-10-11 | 2015-07-28 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| USRE46073E1 (en) | 2001-10-11 | 2016-07-19 | Arkray, Inc. | Method of pre-treating sample for measuring saccharified amine and method of measuring saccharified amine |
| WO2005056823A1 (en) | 2003-12-12 | 2005-06-23 | Arkray, Inc. | Method of measuring saccharified amine |
| US7794966B2 (en) | 2003-12-12 | 2010-09-14 | Arkray, Inc. | Method of measuring glycated amine |
| JP4834771B2 (en) * | 2006-07-21 | 2011-12-14 | エフ．ホフマン−ラ ロシュ アーゲー | Hemoglobin digestion reagent |
| JP2019086450A (en) * | 2017-11-09 | 2019-06-06 | 国立大学法人弘前大学 | Analysis device for whole blood albumin, and analysis method for whole blood albumin |
| JP7015521B2 (en) | 2017-11-09 | 2022-02-03 | 国立大学法人弘前大学 | Whole blood albumin analyzer and whole blood albumin analysis method |
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