Microfluidic Immunoassay System for Rapid Detection and Semi-Quantitative Determination of a Potential Serum Biomarker Mesothelin
Microfluidic Immunoassay System for Rapid Detection and Semi-Quantitative Determination of a Potential Serum Biomarker Mesothelin
复制标题
用于快速检测和半定量测定潜在血清生物标志物间皮素的微流控免疫分析系统。
DOI：
10.1021/acssensors.9b01430
复制
发表时间：
2019-11-01
期刊：
影响因子：
8.9
通讯作者：
Sui, Guodong
中科院分区：
文献类型：
--
作者：
Duan, Xiaoxiao;Zhao, Linlin;Sui, Guodong
间皮素（MSLN）被认为是胰腺癌早期诊断的潜在血清学肿瘤生物标志物。然而，传统检测方法灵敏度低、试剂消耗量大、耗时长，限制了其在临床疾病诊断中的应用。本研究将免疫分析技术与微流控芯片相结合，开发了一种微流控免疫分析系统（MIAS），可用于快速半定量检测血清MSLN水平。MIAS由12个均匀结构组成，包括12个入口、12个反应室和一个出口，可同时测量4个样品，重复3次。一个独特的微阵列圆柱体位于每个反应室的末端，在那里进行免疫测定以捕获微球。当微球填充反应柱时，截留效率可达80%左右。结果表明，荧光强度与MIAS上的MSLN浓度成正比（R-2 = 0.95）。随后，选取上海长海医院8例胰腺癌患者、4例胰腺炎患者、4例其他消化系统疾病患者（胃癌2例、胆管狭窄1例、胆管结石1例）的16例临床血清样本。通过MIAS检测这些样品的MSLN水平。结果表明，MIAS与传统酶联免疫吸附试验（ELISA）的相关系数为0.93。MSLN荧光强度和浓度的检出限与6a.u相近。分别接近20 pg/mL。与ELISA的2小时相比，MIAS的整个分析时间缩短至40分钟。MIAS数据的统计分析显示，胰腺癌中MSLN过表达（P值= 0.0014）。医院中MIAS检测MSLN的诊断准确率为87.5%，ELISA检测CA19-9的诊断准确率为81.3%。MSLN有助于胰腺癌和其他疾病的早期诊断，对胰腺癌和非胰腺癌具有显著的鉴别能力（P值= 0.0159）。本研究结果表明，MIAS具有成为MSLN快速检测和半定量测定的新型血清学肿瘤标志物检测平台的潜力，在临床早期诊断中具有广泛的应用前景。
Mesothelin (MSLN) is considered as a potential serological tumor biomarker for early diagnosis of pancreatic cancer. Nevertheless, low sensibility, high reagent consumption, and time costs of traditional detection methods limit their utility in clinical disease diagnoses. Here, we combined the immunoassay technique with microfluidic chips to develop a microfluidic immunoassay system (MIAS) that can be used for rapid semi-quantitative detection of serum MSLN levels. The MIAS was composed of 12 uniform structures, including 12 inlets, 12 reaction chambers, and one outlet allowing measurement of four samples with three repeats, simultaneously. A unique microarray cylinder was located at the end of each reaction chamber where immunoassay was performed to trap microspheres. A feasible interception efficiency (similar to 80%) was attained, with microspheres filling the reaction column. It has been demonstrated that fluorescence intensity is proportional to the MSLN concentration on the MIAS (R-2 = 0.95). Subsequently, 16 clinical serum samples collected from Changhai Hospital, Shanghai were selected from eight patients with pancreatic cancers, four with pancreatitis, and four with other digestive system diseases (2 gastric cancer, 1 bile duct stricture, and 1 bile duct stones). MSLN levels for these samples were detected via MIAS. The results showed a significant correlation between MIAS and traditional enzymelinked immunosorbent assay (ELISA), with the correlation coefficient, 0.93. The detection limit of MSLN fluorescence intensity and concentration was similar to 6a.u. and similar to 20 pg/mL, respectively. The entire duration of analysis by MIAS decreased to similar to 40 min compared to 2 h by ELISA. Statistical analysis of MIAS data revealed that MSLN was overexpressed in pancreatic cancer than in the others (P value =, 0.0014). Moreover, the diagnostic accuracies of MSLN detected by MIAS and CA19-9 detected by ELISA in hospitals were 87.5 and 81.3%, respectively. MSLN is helpful for the early diagnosis of pancreatic cancer and other diseases, and it had a significant ability to discriminate between pancreatic and nonpancreatic cancers (P value = 0.0159). The results from this study show that MIAS has the potential to become a new serological tumor marker detection platform for rapid detection and semi-quantitative determination of MSLN and would have broad applications in early clinical diagnosis.