Department of Biomedicine and Genomics, Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, Russia.
Int J Mol Sci. 2023 Feb 24;24(5):4483. doi: 10.3390/ijms24054483.
Phage therapy is now seen as a promising way to overcome the current global crisis in the spread of multidrug-resistant bacteria. However, phages are highly strain-specific, and in most cases one will have to isolate a new phage or search for a phage suitable for a therapeutic application in existing libraries. At an early stage of the isolation process, rapid screening techniques are needed to identify and type potential virulent phages. Here, we propose a simple PCR approach to differentiate between two families of virulent phages ( and ) and eleven genera of virulent phages (, , , , , , , , , and ). This assay includes a thorough search of a dataset comprising ( = 269) and ( = 480) phage genomes available in the NCBI RefSeq/GenBank database for specific genes that are highly conserved at the taxonomic group level. The selected primers showed high sensitivity and specificity for both isolated DNA and crude phage lysates, which permits circumventing DNA purification protocols. Our approach can be extended and applied to any group of phages, given the large number of available genomes in the databases.
噬菌体疗法现在被视为克服当前全球多药耐药菌传播危机的一种有前途的方法。然而，噬菌体具有高度的菌株特异性，在大多数情况下，必须分离新的噬菌体或在现有文库中寻找适合治疗应用的噬菌体。在分离过程的早期阶段，需要快速筛选技术来识别和分型潜在的毒性噬菌体。在这里，我们提出了一种简单的 PCR 方法来区分两种毒性噬菌体（和）和十一种毒性噬菌体属（，，，，，，，，，和）。该检测方法包括对 NCBI RefSeq/GenBank 数据库中可用的 269 个噬菌体基因组和 480 个噬菌体基因组数据集进行彻底搜索，以寻找在分类群水平高度保守的特定基因。选择的引物对分离的 DNA 和粗噬菌体裂解物均具有高灵敏度和特异性，这允许绕过 DNA 纯化方案。鉴于数据库中存在大量可用基因组，我们的方法可以扩展并应用于任何噬菌体群体。
