Research of analytical methods for infection test by capillary electrophoresis.

毛细管电泳感染检测分析方法研究。

• 批准号: 11672299
• 负责人: OKABE Hiroaki
• 金额: $ 2.3万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672299/
• 关键词: capillary electrophoresis; serum protein; LD isoenzymes; Alp isoenzymes; M-protein; infection test; HBs antigen; antigen-antibody products; 分析技術; 肝炎

We have developed and evaluated the analytical method for clinical tests (proteins, isoenzymes and infection tests) by capillary electrophoresis (CE). In the infection test of HBs antigen, we established an analysis optimum condition passed through studies such as selection of antibody, selection of running buffer, change of the structure of sample molecule by change of applied voltage, development of analysis personal computer software. Assay conditions as follow : ten μl of 50 times diluted patient serum with distilled water or standard of HBs antigen (Kyokuto pharmaceutical Co., Tokyo, Japan), 10μL of latex banding anti-HBs antigen monoclonal antibody (Kyokuto), 10 μL of anti HBs antigen monoclonal antibody (Wako Pure Chemicals Co. Osaka Japan) and 50 μL of FITC banding anti rat IgG monoclonal antibody (Wako) were mixed. Mixture was introduced by pressure injection for 2 sec (10nL) to 75μm X 20cm uncoated fused capillary column. CE (Paragon CZE 2000 system, Beckman Coluter Inc.Brea, … More USA) was performed at 24℃, 10kV applied for 15 min in 100 mmol/l borate buffer (pH 10.04), HBs antigen activity was monitored by following fluorescence at 480/520. Peak of antigen-antibody reaction product was not detected for HBs antigen negative samples, peak of antigen-antibody reaction products were observed for HBs antigen positive samples approximately 10 min. Results of AXIM method (ABBOTT, USA) and the peak area of proposed method showed good correlation (r=0.94). Also, correlation of standard HBs antigen and results of AXIM method were excellent (r=0.90), however little differences were recognized in lower activity. The reproducibility for the proposed method were excellent (less than CV 2%). Normal concentration of various kinds of anticoagulants, albumin, glucose, ascorbic acid and hemoglobin had no effects on the results. We conclude that proposed HBs antigen assay by CE is easy and precise in clinical laboratory use. Furthermore, we are continue studying for the assay of other infection tests and the assay of lipids tests by CE. Less

我们开发并评价了毛细管电泳（CE）临床试验（蛋白质、同工酶和感染试验）的分析方法。在乙型肝炎病毒表面抗原的感染试验中，通过对抗体的选择、运行缓冲液的选择、外加电压的改变对样品分子结构的影响、分析软件的开发等研究，建立了最佳的分析条件。测定条件如下：10 μl用蒸馏水稀释50倍的患者血清或HBs抗原标准品（Kyokuto Pharmaceutical Co.，Tokyo，Japan）、10μL乳胶显带抗-HBs抗原单克隆抗体（Kyokuto）、10 μL抗-HBs抗原单克隆抗体（和子Pure Chemicals Co. Osaka Japan）和50 μL FITC显带抗大鼠IgG单克隆抗体（和子）混合。通过压力进样2秒（10 nL）将混合物引入75μm X 20 cm未涂层熔融毛细管柱中。CE（Paragon CZE 2000系统，Beckman Coluter Inc.Brea， ...更多信息 USA），在100 mmol/l硼酸盐缓冲液（pH10.04）中，24℃，10 kV电压作用15 min，以480/520的荧光强度监测HBs抗原活性。HBs抗原阴性样品未检测到抗原抗体反应产物峰，HBs抗原阳性样品约10 min出现抗原抗体反应产物峰，阿克西姆法（ABBOTT，USA）结果与拟定方法峰面积相关性良好（r=0.94）。此外，标准HBs抗原与阿克西姆法结果的相关性极好（r=0.90），但在较低的活性方面差异不大。方法的重现性良好（CV <2%）。正常浓度的各种抗凝剂、白蛋白、葡萄糖、抗坏血酸和血红蛋白对结果无影响。我们的结论是，拟议的乙肝表面抗原检测CE是简单和精确的临床实验室使用。此外，我们正在继续研究其他感染试验的测定和CE脂质试验的测定。少

项目成果

宇治義則: "キャピラリー電気泳動臨床検査への応用"日本臨床化学会九州支部会誌. 9・1. 44-51 (1999)

宇治义德：“毛细管电泳在临床检测中的应用”日本临床化学学会九州分会杂志 9・1（1999）。

宇治義則: "キャピラリー電気泳動法の応用"日本臨床化学会関東支部会会誌. 11・1. 6-8 (1999)

宇治义德：“毛细管电泳的应用”日本临床化学学会关东分会杂志11・1（1999）。

Yoshinori Uji and Hiroaki Okabe: "Serum LD isoenzymes by capillary electrophoresis."Indian Journal of Clinical Chemistry 2001. (in Press). (2001)

Yoshinori Uji 和 Hiroaki Okabe：“通过毛细管电泳测定血清 LD 同工酶。”印度临床化学杂志 2001 年。（出版中）。

Uji, Y.and Okabe H.: "Serum protein analysis by capillary elctrophoresis."Indian Journal of Clinical Chemistry. (in press). (2001)

Uji, Y. 和 Okabe H.：“通过毛细管电泳进行血清蛋白分析。”印度临床化学杂志。

Yoshinori Uji and Hiroaki Okabe: "Serum Protein analysis by Capillary Electrophoresis."Indian Journal of Clinical Chemistry 2001. (in Press). (2001)

Yoshinori Uji 和 Hiroaki Okabe：“通过毛细管电泳进行血清蛋白分析”。《印度临床化学杂志》2001 年。（出版中）。