New England Biolabs, Ipswich, MA, USA.
Commun Biol. 2022 Sep 21;5(1):999. doi: 10.1038/s42003-022-03973-x.
Detection of nucleic acid amplification has typically required sophisticated laboratory instrumentation, but as the amplification techniques have moved away from the lab, complementary detection techniques have been implemented to facilitate point-of-care, field, and even at-home applications. Simple visual detection approaches have been widely used for isothermal amplification methods, but have generally displayed weak color changes or been highly sensitive to sample and atmospheric effects. Here we describe the use of pyridylazophenol dyes and binding to manganese ion to produce a strong visible color that changes in response to nucleic acid amplification. This detection approach is easily quantitated with absorbance, rapidly and clearly visible by eye, robust to sample effects, and notably compatible with both isothermal and PCR amplification. Nucleic acid amplification and molecular diagnostic methods are being used in an increasing number of novel applications and settings, and the ability to reliably and sensitively detect them without the need for additional instrumentation will enable even more access to these powerful techniques.
核酸扩增的检测通常需要复杂的实验室仪器，但随着扩增技术逐渐走出实验室，互补的检测技术被应用于即时检测、现场检测，甚至家庭检测。简单的目视检测方法已被广泛应用于等温扩增方法，但通常显色变化较弱，或对样本和大气影响非常敏感。在这里，我们描述了使用吡啶偶氮苯酚染料与锰离子结合，产生对核酸扩增有响应的强可见颜色变化。这种检测方法很容易通过吸光度定量，用肉眼快速、清晰地观察，对样本效应具有较强的稳健性，并且与等温扩增和 PCR 扩增都很兼容。核酸扩增和分子诊断方法正在越来越多的新应用和环境中使用，无需额外仪器即可可靠、灵敏地检测它们的能力将使这些强大的技术得到更广泛的应用。
