The extract from isolated rabbit aortae were added to a Krebs solution for 1 h to secrete enzymes. Then, the arginine amidase activity identified in the obtained sample solution was separated using affinity adsorption on lysine-, anti-thrombin lll- and aprotinin conjugated columns. Enzymes adsorbed on lysine and aprotinin affinity columns were found in the extract of isolated rabbit aortae. On the other hand, few enzymes showing affinity with the anti-thrombin lll column were detected. Two forms of arginine amidases with different affinities were isolated and these showed different substrate specificities. The arginine amidase obtained from the lysine affinity column was supposed to be plasmin because it was completely inhibited by antiplasminogen.

抄録全体を表示

An antibody-lectin sandwich enzyme immunoassay for analysis of altered sugar chains in serum transferrin of patients with hepatoma was developed. Anti-transferrin lgGcoated polystyrene beads incubated with neuraminidase-treated serum samples were reacted with peroxidase (POD)-conjugated lectins such as ricinus communis agglutinin (RCA), lens culinaris agglutinin (LCA), phytohemagglutinin-E₄ (EPHA), and phytohemagglutin-L₄ (LPHA). The amounts of lectins bound to the sugar chains in transferrin were determined by measurement of the POD activities. The LCA/RCA, EPHA/RCA, and LPHA/RCA POD activity ratios were increased significantly in sera from patients with hepatoma in comparison with those in healthy individuals. These results indicated that fucosylation at the reduced end of the N-acetylglucosamine residue, bisecting N-acetylglucosamine at the trimannosyl core, and highly-branched complex-type sugar chains were increased in serum transferrin of patients with hepatoma.

抄録全体を表示

We developed an anti-GOR assay using the Luminomaster TM system, a fully automated, random access chemiluminescent enzyme immunoassay system. GOR antibody, which is often detected in the serum of patients infected with Hepatitis C virus (HCV), is closely associated with HCV infection.
This new assay provides detection of GOR antibody in 45 min, about half the time of existing EIA methods. The standard curve is 0.1-300 GOR unit/ml (1/10th-300 times cut-off value), givinga detection range about 30 times greater than that of existing assays. This greater detection range allows quantitative determination of low-titer samples without the need for additional detectionprocedures. This assay can also be used to evaluate the clinical efficacy of interferon (IFN) therapy in patients with HCV.

抄録全体を表示

ドライケミストリー法は, 面倒な試薬調製が一切不要で, 簡単に検査ができ速やかに結果が得られる測定法で, 方式として多層分析フィルム法と試験紙法が知られている。一方C反応性タンパク質 (CRP) は急性炎症を初期の段階で鋭敏にとらえることができるため, 測定の迅速性、簡便性が強く求められている。我々は血清中のCRPを迅速簡便に測定する多層分析フィルムを開発した。本法は細菌由来のα-アミラーゼで標識した抗ヒトCRPモノクローナル抗体と, 不溶性澱粉を用いることを特徴とする均一系酵素免疫反応に基づき, 展開層, バリア層, 発色試薬層の3層で構成される。専用測定機 (富士ドライケム5500/3000) を用い検討した結果, 検出感度は0-5mg/100m農で精度や従来法との相関性は良好であった。

抄録全体を表示

予防医学事業中央会は小児成人病検診の研究を昭和62年度から実施し, 血清総コレステロール (TC) などを測定している。36支部の全国的規模で行われる検診は, 地域特性の有無を見るために, 測定値は正確さが保証され各施設問で互換性のあることが必須条件である。参加35支部を対象に1988年から1992年まで5年間に毎年1回TCの精度管理調査を実施した。本調査では測定値の施設間差とともに各施設内精密度 (再現性) について解析を試みた。その結果, 年々解析結果に改善が認められ, 施設間差の変動係数 (CV) は2.2～2.6%となった。また凍結乾燥血清は精度管理調査試料として不適切であり, 凍結プール血清が有用であった。指定標準物質は認証標準血清や二次標準血清など正確度の保証された血清を用いることが重要である。正確度の改善には, 各施設において日常使用している標準血清と指定標準血清を用いて同一検体を同時に測定し, それらの結果から補正係数を求めて各支部の測定値を補正することが有効であることを確認した。

抄録全体を表示

硫酸亜鉛混濁試験 (ZTT) は, 血清蛋白成分の量的, 質的異常を観察する簡易的な測定法で重要な検査項目の一つである。しかし標準法の処方は2.6mMのバルビタール緩衝液を使用しているため, 反応に重大な因子となるpHを維持する能力が弱く, 開栓状態では空気中の炭酸ガスを吸収し, 施栓した状態ではその化学的安定性から経日的にpHが低下して感度が上昇し, それが日差変動や施設間差の原因になっていた。そこで解離度が低く, また化学的に安定なビストリスを緩衝剤に用いる新規なクンケル試液について検討した結果, 標準法と良好な相関性が得られ, 施栓, および開栓状態で優れた経日安定性を示した。ZTT測定の臨床的意義の再認識に貢献できると思われる。

抄録全体を表示

The levels of urinary total sulfated bile acids (U-TSBA) in 12 social alcohol drinkers and 20 heavy drinkers with liver diseases were investigated. In comparison with the reference value, no significant elevation of U-TSBA was found in either group regardless of the severity of alcoholic liver injury. Two of the three heavy drinkers with abnormal values of UTSBA were positive for hepatitis C virus antibody and showed corresponding histological changes compatible with viral hepatitis. The results show that alcoholic liver injury per se may have minimal effects, if any, on urinary sulfated bile acid excretion, while other hepatobiliary diseases including viral hepatitis result in significant increases.

抄録全体を表示

A reference method for the determination of creatinine in serum is described. The method is based on the use of high performance liquid chromatography (HPLC), employing phosphate buffer (pH 5.75) as a mobile phase at a flow rate of 0.7 ml/min. A serum sample is deproteinized with trichloroacetic acid solution. Creatinine is separated from coexisting substances in serum by cation-exchange chromatography on Shimadzu Shim-pak ISC 05/S0504 column. The absorbance of creatinine in the eluate is monitored at 234 nm. Sera and reference standards (NIST, SRM914) are assaied in every 7 minutes. The relationship between the concentration of creatinine reference standard and peak area is linear up to 1327.4 μmol/l. The coefficient of variation (CV) is 1.0 to 1.8% at 56.6 to 511.5 μEmol/l of creatinine. Between-day precision studies for the levels of 91.1 and 515.9 μmol/l provided CV of 2.7 and 0.9%, respectivity. The recovery rate of creatinine added to serum was observed in the range 99.6 to 102.0%. None of 24 kinds of endogenous and exogenous substances exerted interferences. It was demonstrated that the alkaline picrate procedure exhibits a statistically significant bias in interlaboratory and survey studies employing this procedure as a reference.

抄録全体を表示

Guideline I. JSCC concensus method. This method was established in order to avoid discrepancies between JSCC recommended methods and routine methods in terms of assay temperature. The assay conditions for the JSCC consensus method are those recommended by JSCC, except that incubation is carried out at 37°C. Guideline II. Reagents prepared according to the recommendation of JSCC. These are reagent kits for automated analyzers which are prepared in such a way that the final concentrations of each reagent in the reaction mixture remain the same as those used by the JSCC recommended method, and preservatives or stabilizing agentsadded to the reagents have no influence on enzyme assays. Guideline III. Quality assurance survey. We are hoping that more institutions will report their results according to this method and that the accuracy of enzyme assays will be controlled by the JSCC consensus method.

抄録全体を表示