Smoldovskaya O V, Voloshin S A, Novikov A A, Aleksandrova E N, Feyzkhanova G U, Rubina A Yu
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.
Moscow Clinical Scientific Center A.S. Loginov, 111123 Moscow, Russia.
Mol Biol. 2022;56(2):290-296. doi: 10.1134/S0026893322020145. Epub 2022 Apr 14.
Serum amyloid A is an inflammatory biomarker whose concentration changes during infectious and inflammatory diseases. SAA's tendency for aggregation and complex formation makes it difficult to determine its concentration in samples, especially when there is an increased level of it. Immunofluorescence SAA determination on a microarray was adapted for SAA quantification in human serum. Both the procedure and the diluent for the calibrator samples were chosen to obtain a dynamic range between 1 and 100 μg/mL. Mixtures of animal (rabbit, goat, mouse) sera with recombinant antigen diluted in certain concentrations were used for the calibrator samples. The method was tested using serum samples from 15 patients with rheumatoid arthritis or ankylosing spondylitis and 9 healthy donors. The results obtained on the microarray demonstrated a good correlation with the results determined by ELISA (Pearson's correlation coefficient is 0.93). The method developed could be a convenient tool for assessing SAA levels in a number of diseases, such as rheumatoid arthritis or infections of various etiologies, characterized by a significant increase in the level of this protein in the blood. The use of a microarray for the analysis allows the determination of the SAA concentration simultaneously with other inflammatory biomarkers.
血清淀粉样蛋白A是一种炎症生物标志物,其浓度在感染性疾病和炎症性疾病期间会发生变化。血清淀粉样蛋白A的聚集和复合物形成倾向使得难以测定其在样本中的浓度,尤其是当其水平升高时。微阵列上的免疫荧光血清淀粉样蛋白A测定法适用于人血清中血清淀粉样蛋白A的定量分析。校准样本的程序和稀释剂均经过选择,以获得1至100μg/mL的动态范围。校准样本使用动物(兔、山羊、小鼠)血清与以特定浓度稀释的重组抗原的混合物。该方法使用来自15名类风湿性关节炎或强直性脊柱炎患者和9名健康供体的血清样本进行了测试。在微阵列上获得的结果与酶联免疫吸附测定法(ELISA)测定的结果显示出良好的相关性(皮尔逊相关系数为0.93)。所开发的方法可能是评估多种疾病中血清淀粉样蛋白A水平的便捷工具,这些疾病如类风湿性关节炎或各种病因的感染,其特征是血液中该蛋白水平显著升高。使用微阵列进行分析能够同时测定血清淀粉样蛋白A浓度和其他炎症生物标志物。