Korkaew Pratchayaporn, Vilaivan Tirayut
Organic Synthesis Research Unit, Department of Chemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Patumwan, Bangkok 10330, Thailand.
Nucleic Acids Symp Ser (Oxf). 2008(52):251-2. doi: 10.1093/nass/nrn127.
Peptide nucleic acid (PNA) is a new class of DNA mimic, which is potentially useful for diagnostic and therapeutic applications due to its high affinity and specificity of binding to target nucleic acids. The goal of this work is to develop a simple and sensitive fluorimetric-based detection of DNA sequences using new fluorescent labeled PNA probes previously developed in our laboratory. The method relies on the preferential absorption of PNA.DNA hybrid (negatively charged) over unhybridized PNA (neutral) on an anion exchange solid support. This enables detection of the PNA-nucleic acid interactions, which will take place only when the PNA and DNA sequences are fully complementary directly on the solid support by fluorescence microscopy. With the high specificity of this new PNA-based technology, single mismatches in 9-13 bases target DNA sequences were readily distinguished. Based on this principle, a prototype for detection of single nucleotide polymorphisms (SNP) without the need for labeling the DNA substrates has been developed.
肽核酸（PNA）是一类新型的DNA模拟物，由于其与靶核酸结合具有高亲和力和特异性，因而在诊断和治疗应用方面具有潜在用途。本研究的目的是利用我们实验室之前开发的新型荧光标记PNA探针，开发一种基于荧光法的简单且灵敏的DNA序列检测方法。该方法依赖于在阴离子交换固体支持物上，PNA-DNA杂交体（带负电荷）比未杂交的PNA（呈中性）具有优先吸附性。这使得能够检测PNA与核酸的相互作用，只有当PNA和DNA序列完全互补时，这种相互作用才会在固体支持物上直接通过荧光显微镜发生。基于这种新型基于PNA的技术的高特异性，9至13个碱基的靶DNA序列中的单个错配很容易被区分出来。基于这一原理，已经开发出一种无需标记DNA底物即可检测单核苷酸多态性（SNP）的原型。
