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采用一种廉价、灵敏且简单的夹心酶联免疫吸附测定技术联合检测铁蛋白、可溶性转铁蛋白受体、视黄醇结合蛋白和C反应蛋白。

Combined measurement of ferritin, soluble transferrin receptor, retinol binding protein, and C-reactive protein by an inexpensive, sensitive, and simple sandwich enzyme-linked immunosorbent assay technique.

作者信息

Erhardt Juergen G, Estes John E, Pfeiffer Christine M, Biesalski Hans K, Craft Neal E

机构信息

SEAMEO, Regional Center for Community Nutrition, University of Indonesia, Salemba Raya 6, Jakarta 10430, Indonesia.

出版信息

J Nutr. 2004 Nov;134(11):3127-32. doi: 10.1093/jn/134.11.3127.

Abstract

The measurement of vitamin A (VA) and iron status is very important in the assessment of nutritional deficiencies. The objective of this research was to develop a sandwich ELISA technique for the simultaneous measurement of ferritin, soluble transferrin receptor, retinol binding protein, and C-reactive protein (CRP) as indicators for VA and iron status. The inclusion of CRP as marker of infection allows for more accurate interpretation of VA and iron status. This is accomplished in a 30-microL serum or plasma sample using an ELISA with different capture and detection antibodies and different dilutions of the sample. Commercially available clinical serum controls were used for calibration purposes. The developed assays were compared to commercially available traditional tests. Regression coefficients comparing both assays were better than 0.84 (P < 0.001). Using a limited sample set, the sandwich ELISA assay produced very similar specificity and sensitivity compared to traditional methods when common cutoff values were applied. Intra- and interassay variability was between 5 and 14% for all tests. The cost of the materials for all 5 measurements decreases to less than $1/sample if a large number of samples is analyzed. Due to the low cost, high throughput, and comparability to traditional tests, this procedure has several advantages for assessing VA and iron status in population surveys.

摘要

维生素A(VA)和铁状态的测定在营养缺乏评估中非常重要。本研究的目的是开发一种夹心ELISA技术,用于同时测定铁蛋白、可溶性转铁蛋白受体、视黄醇结合蛋白和C反应蛋白(CRP),作为VA和铁状态的指标。将CRP作为感染标志物纳入,有助于更准确地解读VA和铁状态。这通过使用具有不同捕获和检测抗体以及不同样品稀释度的ELISA,在30微升血清或血浆样本中完成。使用市售临床血清对照进行校准。将开发的检测方法与市售传统检测方法进行比较。两种检测方法的回归系数优于0.84(P < 0.001)。当应用常见临界值时,使用有限样本集,夹心ELISA检测与传统方法相比产生非常相似的特异性和灵敏度。所有检测的批内和批间变异在5%至14%之间。如果分析大量样本,所有5项测量的材料成本降至低于1美元/样本。由于成本低、高通量且与传统检测方法具有可比性,该方法在人群调查中评估VA和铁状态具有多个优点。

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