当前位置：
X-MOL 学术
›
Anal. Methods
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Development of immunomagnetic droplet-based digital immuno-PCR for the quantification of prostate specific antigen†
Analytical Methods ( IF 2.7 ) Pub Date : 2018-07-02 00:00:00 , DOI: 10.1039/c8ay00921j Wenhan Zhou 1, 2, 3, 4
Analytical Methods ( IF 2.7 ) Pub Date : 2018-07-02 00:00:00 , DOI: 10.1039/c8ay00921j Wenhan Zhou 1, 2, 3, 4
Affiliation
|
Immuno-PCR (IPCR) is an immunoassay that employs DNA as the signal generator and utilizes both the versatility of antigen–antibody interactions and the exponential amplification power of PCR. Droplet-based digital PCR (ddPCR) is the third and latest generation of PCR technology, and the integration of ddPCR into IPCR will hopefully enable the absolute quantification and highly sensitive detection of proteins. However, the lack of practical washing methods in the microfluidic droplet system still remains a huge obstacle for such integration. Here, we report a novel droplet-based digital immuno-PCR (ddIPCR) method with magnetic beads as the solid phase. Supported by their separate functions with and without a magnetic field, the employment of magnetic beads successfully builds a bridge between IPCR and droplet microfluidics. Using prostate specific antigen (PSA) as a model antigen, the established immunomagnetic ddIPCR method could detect PSA down to 0.48 ng mL−1 with a linear range from 0.5 to 30 ng mL−1. Also, the detection of human serum samples by immunomagnetic ddIPCR correlated well with the clinical results. The immunomagnetic ddIPCR method provides a novel and highly sensitive detection route for the quantification of antigens, showing great potential in protein research and clinical diagnosis.
中文翻译：
开发基于免疫磁珠的数字免疫PCR定量前列腺特异性抗原†
免疫PCR（IPCR）是一种利用DNA作为信号发生器的免疫测定，既利用了抗原-抗体相互作用的多功能性，又利用了PCR的指数扩增能力。基于液滴的数字PCR（ddPCR）是第三代也是最新一代的PCR技术，将ddPCR集成到IPCR中有望实现蛋白质的绝对定量和高度灵敏的检测。然而，在微流体液滴系统中缺乏实用的洗涤方法仍然是这种整合的巨大障碍。在这里，我们报告一种新颖的基于液滴的数字免疫PCR（ddIPCR）方法，其中磁珠为固相。在有磁场和无磁场的情况下，借助其各自的功能，磁珠的使用成功地在IPCR和液滴微流体之间架起了一座桥梁。-1的线性范围是0.5到30 ng mL -1。同样，通过免疫磁ddIPCR检测人血清样品与临床结果也有很好的相关性。免疫磁ddIPCR方法为抗原定量提供了一种新颖且高度灵敏的检测途径，在蛋白质研究和临床诊断中显示出巨大的潜力。
更新日期：2018-07-02
中文翻译：
开发基于免疫磁珠的数字免疫PCR定量前列腺特异性抗原†
免疫PCR（IPCR）是一种利用DNA作为信号发生器的免疫测定，既利用了抗原-抗体相互作用的多功能性，又利用了PCR的指数扩增能力。基于液滴的数字PCR（ddPCR）是第三代也是最新一代的PCR技术，将ddPCR集成到IPCR中有望实现蛋白质的绝对定量和高度灵敏的检测。然而，在微流体液滴系统中缺乏实用的洗涤方法仍然是这种整合的巨大障碍。在这里，我们报告一种新颖的基于液滴的数字免疫PCR（ddIPCR）方法，其中磁珠为固相。在有磁场和无磁场的情况下，借助其各自的功能，磁珠的使用成功地在IPCR和液滴微流体之间架起了一座桥梁。-1的线性范围是0.5到30 ng mL -1。同样，通过免疫磁ddIPCR检测人血清样品与临床结果也有很好的相关性。免疫磁ddIPCR方法为抗原定量提供了一种新颖且高度灵敏的检测途径，在蛋白质研究和临床诊断中显示出巨大的潜力。
京公网安备 11010802027423号