Joda Hamdi, Beni Valerio, Katakis Ioanis, O'Sullivan Ciara K
INTERFIBIO Research Group, Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, 43007 Tarragona, Spain.
INTERFIBIO Research Group, Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, 43007 Tarragona, Spain; Institucio Catalana de Recerca i Estudis Avançats, Passeig Lluis Companys 23, 08010 Barcelona, Spain.
Anal Biochem. 2015 Apr 1;474:66-8. doi: 10.1016/j.ab.2014.11.021. Epub 2014 Dec 15.
We report on a simple approach to enhance solid-phase hybridization-based single base mismatch discrimination at high ionic strength based on the deliberate insertion of a natural DNA base mismatch in the surface-tethered probe. A large drop in hybridization signal of single base mismatched alleles using the designed probe as compared with the conventional probe, from 80% to less than 25% of the signal obtained with the fully complementary, non-mutation-containing sequence, when using colorimetric detection was further improved to 20% when using electrochemical detection, attributable to a difference of spacing of immobilized probes. Finally, the designed probe was used for the electrochemical detection of the DQA1*05:05 allele amplified from real human blood samples.
我们报道了一种基于在表面连接的探针中故意插入天然DNA碱基错配，以增强在高离子强度下基于固相杂交的单碱基错配识别的简单方法。使用设计的探针时，与传统探针相比，单碱基错配等位基因的杂交信号大幅下降，在使用比色检测时，从与完全互补、无突变序列获得的信号的80%降至不到25%，而在使用电化学检测时进一步提高到20%，这归因于固定化探针间距的差异。最后，设计的探针用于对从真实人类血液样本中扩增的DQA1*05:05等位基因进行电化学检测。
