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首页> 美国卫生研究院文献>Journal of Clinical Laboratory Analysis >Development of a microplate ELISA for circulating E‐selectin: assay characterization comparison with a commercial kit wand establishment of normal reference values
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Development of a microplate ELISA for circulating E‐selectin: assay characterization comparison with a commercial kit wand establishment of normal reference values

机译：用于循环E-选择素的微孔板ELISA的开发：测定表征与市售试剂盒进行比较通过棒建立正常参考值

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E‐selectin, an adhesion molecule, is detectable in the blood. Serum levels of E‐selectin can be a prognostic indicator for various malignancies. We developed a microplate enzyme‐linked immunosorbent assay (ELISA) for the detection of circulating E‐selectin by coating the microwell with monoclonal anti‐E‐selectin. We incubated the specimen and the biotinylated detecting antibody simultaneously, and used HPR‐conjugated streptavidin to generate a signal for quantification. The assay has a sensitivity of 3.8 ng/mL, and the coefficients of variation (CVs) for both within‐day and day‐to‐day precision were all <10%. Our in‐house‐made kit also compared well with a commercial kit from R&D Systems (r=0.95, slope=0.94). Using our kit we determined a normal reference value for Chinese individuals of different ages (30–80 years). We found no significant difference between females and males; however, age appeared to have an impact on the normal E‐selectin value. We found that normal individuals over 60 years old had higher levels of circulating E‐selectin (65.8±19.5 ng/mL, N=111) compared to those under 60 years old (59.5±18.1 ng/mL; =0.002). Our kit appears to have sufficient sensitivity for detecting elevated circulating E‐selectin in various carcinomas. We believe that with the established normal reference value, our in‐house‐developed ELISA kit is well suited for routine clinical laboratory use. J. Clin. Lab. Anal. 17:97–101, 2003. © 2003 Wiley‐Liss, Inc.
机译：E-选择素是一种粘附分子，可在血液中检测到。血清E-选择素水平可以作为各种恶性肿瘤的预后指标。我们开发了一种微孔板酶联免疫吸附测定（ELISA），通过在微孔上涂单克隆抗E-选择素来检测循环中的E-选择素。我们同时孵育了标本和生物素化的检测抗体，并使用了HPR偶联的抗生蛋白链菌素来产生定量信号。该测定法的灵敏度为3.8 ng / mL，日内和每日精度的变异系数（CV）均<10％。我们的内部套件也可以与R＆D Systems的商业套件（r = 0.95，斜率= 0.94）进行比较。使用我们的试剂盒，我们确定了不同年龄（30-80岁）的中国人的正常参考值。我们发现女性与男性之间没有显着差异;但是，年龄似乎对正常的E-选择素值有影响。我们发现60岁以上的正常人的循环E-选择素水平（65.8±19.5 ng / mL，N = 111）高于60岁以下的人（59.5±18.1 ng / mL; = 0.002）。我们的试剂盒似乎具有足够的灵敏度，可检测各种癌症中循环E-选择素的升高。我们相信，使用已建立的正常参考值，我们内部开发的ELISA试剂盒非常适合常规临床实验室使用。 J.临床实验室肛门17：97-101，2003.©2003 Wiley-Liss，Inc.

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