Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas.
Center for Phage Technology, Texas A&M University, College Station, Texas.
Biotechnol Bioeng. 2018 Jun;115(6):1427-1436. doi: 10.1002/bit.26573. Epub 2018 Mar 8.
Rapid identification of specific bacterial strains within clinical, environmental, and food samples can facilitate the prevention and treatment of disease. Fluorescent nanodiamonds (FNDs) are being developed as biomarkers in biology and medicine, due to their excellent imaging properties, ability to accept surface modifications, and lack of toxicity. Bacteriophages, the viruses of bacteria, can have exquisite specificity for certain hosts. We propose to exploit the properties of FNDs and phages to develop phages conjugated with FNDs as long-lived fluorescent diagnostic reagents. In this study, we develop a simple procedure to create such fluorescent probes by functionalizing the FNDs and phages with streptavidin and biotin, respectively. We find that the FND-phage conjugates retain the favorable characteristics of the individual components and can discern their proper host within a mixture. This technology may be further explored using different phage/bacteria systems, different FND color centers and alternate chemical labeling schemes for additional means of bacterial identification and new single-cell/virus studies.
在临床、环境和食品样本中快速鉴定特定的细菌菌株有助于疾病的预防和治疗。由于荧光纳米金刚石 (FND) 具有出色的成像特性、能够接受表面修饰以及没有毒性,因此它们正在被开发为生物学和医学中的生物标志物。噬菌体是细菌的病毒,对某些宿主具有极高的特异性。我们建议利用 FND 和噬菌体的特性,开发与 FND 结合的噬菌体作为长寿命荧光诊断试剂。在这项研究中,我们开发了一种简单的方法,通过分别用链霉亲和素和生物素对 FND 和噬菌体进行功能化,来创建这种荧光探针。我们发现 FND-噬菌体缀合物保留了各个成分的有利特性,并能在混合物中辨别其合适的宿主。这项技术可以使用不同的噬菌体/细菌系统、不同的 FND 色心和替代的化学标记方案进一步探索,以提供细菌鉴定的新方法和用于单细胞/病毒的研究。