文中所述的MPRP方法是将灵敏的多重real time PCR方法与滚环式DNA扩增方法相结合，解决了很多其他基于PCR的SNP检测方法中存在的灵敏度低、通量小、复杂的操作以及高成本等问题，同时实现了便捷地多重real time PCR检测。文中将MPRP方法与三通道微滴芯片式数字PCR平台结果进行比较，充分表明该方法在对靶向药物的患者筛选和耐药性预测中具备显著的应用前景。

数字PCR测定的2D图 ↑

红圈中的点数表示阳性微滴的数量

（A）患者P6的EGFR L858R测定的数字PCR结果

（B）患者P1的EGFR T790M测定的数字PCR结果

          随着靶向药物的持续开发和应用，需建立非入侵性诊断方法以便筛选可进行治疗的患者。而且多个肿瘤相关突变检测对于治疗和耐药性预测意义重大。虽然有很多检测SNP的PCR方法，但是由于灵敏度低、通量小、复杂的操作以及高成本等问题，应用严重受限。为解决上述问题，本文描述的MPRP方法，将灵敏的多重real time PCR方法与滚环式DNA扩增方法进行了有效结合。提高了SNP检测的特异性和灵敏度，同时实现便捷地多重real time PCR检测。在8位患者样本的MPRP方法与数字PCR方法的检测结果比较中，充分表明MPRP方法具有潜在的临床应用前景。同时可以看出，MPRP方法是满足肿瘤相关突变多重检测的特异性和灵敏度要求且便捷的方法。

With the continuous development and application of targeted drugs, it is particularly desirable to find a non- invasive diagnostic approach to screen patients for precision treatment. Specifically, detection of multiple cancer-related mutations is very important for targeted therapy and prediction of drug resistance. Although numerous advanced PCR methods have been developed to discriminate single nucleotide polymorphisms, their drawbacks significantly limit their application, such as low sensitivity and throughput, complicated operations, and expensive costs. In order to overcome these challenges, in this study, we  developed a method combining multiplex and sensitive real-time PCR assay with rolling circle amplification. This allows specific and sensitive discrimination of the single nucleotide mutation and provides convenient multiplex detection by real-time PCR assay. The clinical potential of the MPRP assay was further demonstrated by comparing samples from 8 patients with a digital PCR assay. The coincident results between these two methods indicated that the MPRP assay can provide a specific, sensitive, and convenient method for multiplex detection of cancer-related mutations.