Gong Jian, Li Yishuai, Lin Ting, Feng Xiaoyan, Chu Li
Hebei Medical University Hebei 050017 China
Department of Thoracic Surgery, Hebei Chest Hospital Hebei 050000 China
RSC Adv. 2018 Aug 1;8(48):27375-27381. doi: 10.1039/c8ra05259j. eCollection 2018 Jul 30.
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.
随着靶向药物的不断发展与应用，找到一种非侵入性诊断方法来筛选患者进行精准治疗变得尤为迫切。具体而言，检测多种癌症相关突变对于靶向治疗和耐药性预测非常重要。尽管已经开发出许多先进的聚合酶链反应（PCR）方法来鉴别单核苷酸多态性，但其缺点显著限制了它们的应用，如灵敏度和通量低、操作复杂以及成本高昂。为了克服这些挑战，在本研究中，我们开发了一种将多重灵敏实时PCR检测与滚环扩增相结合的方法。这允许对单核苷酸突变进行特异性和灵敏的鉴别，并通过实时PCR检测提供便捷的多重检测。通过将8例患者的样本与数字PCR检测进行比较，进一步证明了MPRP检测的临床潜力。这两种方法的一致结果表明，MPRP检测可为癌症相关突变的多重检测提供一种特异性、灵敏且便捷的方法。
