Detection of nucleic acids is crucial to the study of their basic properties and consequently to applying this knowledge to the determination of pathologies such as cancer. In this work, our goal is to determine new trends for creating diagnostic tools for cancer driver mutations. Herein, we study a library of natural and modified oligonucleotide duplexes by a combination of optical and theoretical methods. We report a profound effect of additives on the duplexes, including nucleic acids as an active crowder. Unpredictably and inconsistent with DNA+LNA/RNA duplexes, locked nucleic acids contribute poorly to mismatch discrimination in the DNA+LNA/DNA duplexes. We develop a theoretical framework that explains poor mismatch discrimination in KRAS oncogene. We implement our findings in a bead-bait genotyping assay to detect mutated human cancer RNA. The performance of rationally designed probes in this assay is superior to the LNA-primer polymerase chain reaction, and it agrees with sequencing data.

核酸的检测对于研究其基本特性至关重要，因此对于将这些知识应用于确定病理学（例如癌症）至关重要。在这项工作中，我们的目标是确定为癌症驱动突变创建诊断工具的新趋势。在此，我们通过光学和理论方法的结合研究了天然和修饰的寡核苷酸双链体的文库。我们报告了添加剂对双链体的深远影响，包括核酸作为活跃的拥挤者。不可预测且与 DNA+LNA/RNA 双链体不一致，锁核酸对 DNA+LNA/DNA 双链体中的错配辨别贡献很小。我们开发了一个理论框架来解释KRAS中较差的错配歧视致癌基因。我们在珠饵基因分型测定中实施我们的发现，以检测突变的人类癌症 RNA。合理设计的探针在该测定中的性能优于LNA-引物聚合酶链反应，并且与测序数据一致。