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首页> 外文期刊>Journal of Clinical Microbiology >Alkaline phosphatase capture test for the rapid identification of Escherichia coli and Shigella species based on a specific monoclonal antibody.
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Alkaline phosphatase capture test for the rapid identification of Escherichia coli and Shigella species based on a specific monoclonal antibody.

机译：碱性磷酸酶捕获测试，可基于特定的单克隆抗体快速鉴定大肠杆菌和志贺氏菌。

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摘要

A specific monoclonal antibody for Escherichia coli and Shigella sp. alkaline phosphatase was used in an immunocapture assay and allowed identification of E. coli either in culture isolates or directly in clinical specimens. The assay was easy and required only four steps: (i) alkaline phosphatase was released within 10 min by using a gentle lysis procedure, (ii) cell lysates were transferred to antibody-coated tubes for 45 min, (iii) p-nitrophenyl phosphate substrate was added, and (iv) alkaline phosphatase activity was detected in a microsample spectrophotometer at 410 nm. This immunocapture assay was highly specific: only one false-positive reaction was observed with a Klebsiella pneumoniae lysate among the 205 non-E. coli strains tested. The assay was sensitive, detecting 10(7) CFU/ml from culture isolates or 10(5) CFU/ml from urine specimens which had first been grown in phosphate-limiting medium for 2 h. At these bacterial concentrations, the percentages of detected E. coli were high: 91% for blood cultures, 95.4% for culture isolates, and 96.8% for urine specimens.
机译：一种针对大肠杆菌和志贺氏菌的特异性单克隆抗体。碱性磷酸酶用于免疫捕获测定，可用于在培养分离物中或直接在临床样品中鉴定大肠杆菌。该测定很容易且仅需四个步骤：（i）使用温和的裂解程序在10分钟内释放碱性磷酸酶;（ii）将细胞裂解液转移至抗体包被的试管中45分钟;（iii）对硝基苯基磷酸酯添加底物，并且（iv）在微量样品分光光度计中在410nm处检测碱性磷酸酶活性。该免疫捕获测定法具有高度特异性：在205个非E型患者中，肺炎克雷伯菌的裂解物仅观察到一个假阳性反应。大肠杆菌菌株测试。该测定法灵敏，从培养物分离物中检测到10（7）CFU / ml，从尿液样品中检测到10（5）CFU / ml，尿液样品首先在磷酸盐限制培养基中生长2小时。在这些细菌浓度下，检出的大肠杆菌百分比很高：血液培养物为91％，培养物分离物为95.4％，尿液标本为96.8％。

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