Laboratory Corporation of America Holdings, Burlington, NC 27215, USA.
Laboratory Corporation of America Holdings, New Brighton, MN 55112, USA.
Viruses. 2022 Aug 10;14(8):1748. doi: 10.3390/v14081748.
Bacteriophages have been investigated for clinical utility, both as diagnostic tools and as therapeutic interventions. In order to be applied successfully, a detailed understanding of the influence of the human matrix on the interaction between bacteriophage and the host bacterium is required. In this study, a cocktail of luciferase bacteriophage reporters was assessed for functionality in a matrix containing human serum and spiked with . The inhibition of signal and loss of sensitivity was evident with minimal amounts of serum. This phenotype was independent of bacterial growth and bacteriophage viability. Serum-mediated loss of signal was common, albeit not universal, among strains. Immunoglobulin G was identified as an inhibitory component and partial inhibition was observed with both the f(ab') and Fc region. A modified bacteriophage cocktail containing recombinant protein A was developed, which substantially improved signal without the need for additional sample purification. This study highlights the importance of assessing bacteriophage activity in relevant host matrices. Furthermore, it identifies an effective solution, recombinant protein A, for promoting bacteriophage-based detection of in matrices containing human serum.
噬菌体已被研究用于临床应用,既可作为诊断工具,也可作为治疗干预手段。为了成功应用,需要详细了解人体基质对噬菌体与宿主细菌相互作用的影响。在这项研究中,评估了包含荧光素酶噬菌体报告基因的混合物在含有人血清并添加 的基质中的功能。信号抑制和灵敏度降低的现象在最小量的血清存在时就很明显。这种表型与细菌生长和噬菌体活力无关。在 菌株中,血清介导的信号丢失是常见的,但并非普遍存在。免疫球蛋白 G 被鉴定为抑制性成分,并且在 f(ab') 和 Fc 区域都观察到部分抑制。开发了一种含有重组蛋白 A 的改良噬菌体混合物,该混合物在不需要额外样品纯化的情况下,大大提高了信号。这项研究强调了在相关宿主基质中评估噬菌体活性的重要性。此外,它还确定了一种有效的解决方案,即重组蛋白 A,用于促进基于噬菌体的检测在含有人血清的基质中 。