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Aptamer-based fluorometric lateral flow assay for creatine kinase MB
Microchimica Acta ( IF 5.7 ) Pub Date : 2018-07-07 , DOI: 10.1007/s00604-018-2905-4
Jing Zhang , Xuefei Lv , Wei Feng , Xiaoqiong Li , Kunjie Li , Yulin Deng

AbstractA group of aptamers possessing high specificity and affinity for creatine kinase MB (CKMB) was obtained by magnetic systematic evolution of ligands by exponential enrichment. Two aptamers (referred to as C.Apt.21 and C.Apt.30) were found to possess adequately low Kd values. They form a well suited pair for CKMB binding. By using fluorescent microspheres, an aptamer-based lateral flow assay was developed. It is portable, economical, and sensitive. The limit of detection for CKMB is as low as 0.63 ng·mL-1, and the assay works in the 0.005 - 2 μg·mL-1 CKMB concentration range. The method is specific for CKMB, and biomarkers for AMI (such as cardiac troponin I and myoglobin) and serum do not interfere. The strip is highly accurate as shown by analysis of spiked serum samples which gave recoveries ranging between 88 and 117%. Graphical AbstractSchematic of the test strip and sandwich aptamer-based fluorometric lateral flow assay for creatine kinease. The detection is based on the specific affinity between CKMB and selected aptamers to form a sandwich structure.

中文翻译:

基于适体的肌酸激酶 MB 荧光横向流动测定

摘要 通过指数富集的配体磁性系统进化,获得了一组对肌酸激酶MB(CKMB)具有高特异性和亲和力的适配体。发现两个适体(称为 C.Apt.21 和 C.Apt.30)具有足够低的 Kd 值。它们形成了非常适合 CKMB 绑定的一对。通过使用荧光微球,开发了一种基于适体的侧向流动测定。它便携、经济且灵敏。CKMB 的检测限低至 0.63 ng·mL-1,该检测在 0.005 - 2 μg·mL-1 CKMB 浓度范围内有效。该方法对 CKMB 具有特异性,AMI 的生物标志物(如心肌肌钙蛋白 I 和肌红蛋白)和血清不会产生干扰。对加标血清样品的分析表明,该试纸具有高度的准确度,其回收率介于 88% 和 117% 之间。图形摘要肌酸激酶测试条和夹心适体荧光横向流动测定的示意图。检测是基于 CKMB 与选定的适体之间的特异性亲和力形成夹心结构。
更新日期:2018-07-07
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