Defence Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, 117510 Singapore, Singapore.
J Virol Methods. 2010 Aug;167(2):172-7. doi: 10.1016/j.jviromet.2010.04.001. Epub 2010 Apr 23.
Dengue virus causes frequent and cyclical epidemics throughout the tropics, resulting in significant morbidity and mortality rates. There is neither a specific antiviral treatment nor a vaccine to prevent epidemic transmission. The lack of a detailed understanding of the pathogenesis of the disease complicates these efforts. The development of methods to probe the interaction between the virus and host cells would thus be useful. Direct fluorescence labelling of virus would facilitate the visualization of the early events in virus-cell interaction. This report describes a simple method of labelling of dengue virus with Alexa Fluor succinimidyl ester dye dissolved directly in the sodium bicarbonate buffer that yielded highly viable virus after labelling. Alexa Fluor dyes have superior photostability and are less pH-sensitive than the common dyes, such as fluorescein and rhodamine, making them ideal for studies on cellular uptake and endosomal transport of the virus. The conjugation of Alexa Fluor dye did not affect the recognition of labelled dengue virus by virus-specific antibody and its putative receptors in host cells. This method could have useful applications in virological studies.
登革热病毒在热带地区频繁且周期性地引发流行，导致发病率和死亡率居高不下。目前既没有针对该病毒的特效抗病毒治疗方法，也没有可用于预防疫情传播的疫苗。由于对疾病发病机制缺乏深入了解，这些努力的成效受到了影响。因此，开发用于探究病毒与宿主细胞相互作用的方法将很有意义。直接对病毒进行荧光标记有助于可视化病毒与宿主细胞相互作用的早期事件。本报告介绍了一种简便的方法，即用溶解于碳酸氢钠缓冲液中的 Alexa Fluor 琥珀酰亚胺酯染料对登革热病毒进行标记，结果表明标记后病毒仍具有高活力。与常见的荧光染料（如荧光素和罗丹明）相比，Alexa Fluor 染料具有更好的光稳定性和更低的 pH 敏感性，非常适合用于研究病毒在细胞内的摄取和内体运输。Alexa Fluor 染料的标记不影响标记后的登革热病毒被病毒特异性抗体和宿主细胞中假定的受体识别。这种方法在病毒学研究中可能具有广泛的应用。
