Ahn Jeong Jin, Lee Seung Yong, Hong Ji Young, Kim Youngjoo, Kim Gi Won, Hwang Seung Yong
Dept. of Bio-Nanotechnology, Hanyang University, Sangnok-gu, Ansan, Gyeonggi-do, Korea.
Dept. of Molecular and Life Science, Hanyang University, Sangnok-gu, Ansan, Gyeonggi-do, Korea.
Biotechnol Prog. 2015 May-Jun;31(3):730-5. doi: 10.1002/btpr.2054. Epub 2015 Feb 18.
Peptide nucleic acid (PNA) is an artificially synthesized polymer. PNA oligomers show greater specificity in binding to complementary DNAs. Using this PNA, fluorescence melting curve analysis (FMCA) for dual detection was established. Genomic DNA of Mycoplasma fermentans and Mycoplasma hyorhinis was used as a template DNA model. By using one PNA probe, M. fermentans and M. hyorhinis could be detected and distinguished simultaneously in a single tube. The developed PNA probe is a dual-labeled probe with fluorescence and quencher dye. The PNA probe perfectly matches the M. fermentans 16s rRNA gene, with a melting temperature of 72°C. On the other hand, the developed PNA probe resulted in a mismatch with the 16s rRNA gene of M. hyorhinis, with a melting temperature of 44-45°C. The melting temperature of M. hyorhinis was 27-28°C lower than that of M. fermentans. Due to PNA's high specificity, this larger melting temperature gap is easy to create. FMCA using PNA offers an alternative method for specific DNA detection.
肽核酸（PNA）是一种人工合成的聚合物。PNA寡聚物在与互补DNA结合时表现出更高的特异性。利用这种PNA，建立了用于双重检测的荧光熔解曲线分析（FMCA）。发酵支原体和猪鼻支原体的基因组DNA被用作模板DNA模型。通过使用一种PNA探针，可以在单个试管中同时检测和区分发酵支原体和猪鼻支原体。所开发的PNA探针是一种带有荧光和淬灭染料的双标记探针。该PNA探针与发酵支原体16s rRNA基因完全匹配，熔解温度为72°C。另一方面，所开发的PNA探针与猪鼻支原体的16s rRNA基因不匹配，熔解温度为44-45°C。猪鼻支原体的熔解温度比发酵支原体低27-28°C。由于PNA具有高特异性，很容易产生这种较大的熔解温度差异。使用PNA的FMCA为特定DNA检测提供了一种替代方法。
