中文摘要：全球越来越多的耐药细菌感染激发了人们对噬菌体疗法的兴趣。该疗法使用由高度特异性细菌病毒组成的个性化混合物，需要首先在琼脂培养基上进行一系列测试，确定特定细菌靶上最活跃的噬菌体（噬菌体敏感性测试），或从环境样品中分离出新的裂解噬菌体（噬菌体库的富集）。然而，这些基于培养的测试技术主要通过直接目测噬菌斑进行判断。本研究旨在通过计算机辅助的方法，简化和加速噬菌体治疗的诊断，并研究噬菌斑的生长动力学。为此，专门设计了宽视场无透镜成像装置，可以通过超大面积传感器(3.3cm2)进行连续监测。本文报道了细菌对金黄色葡萄球菌噬菌体在3小时内的敏感性，并在8小时20分钟内评估了感染滴度。肉眼观察和噬菌斑计数仍是噬菌体治疗前广泛使用的易感性检测技术，耗时12-24小时，而本研究提供的方法大大缩短了检测时间。此外，通过持续监测样品，可以研究菌斑的生长动力学，从而更深入地了解噬菌体和细菌之间的相互作用。最后，借助4.3mu m的分辨率，在噬菌斑的边界内检测到肺炎克雷伯菌的噬菌体耐药细菌微菌落，表明该装置也适合用于追踪噬菌体耐药性。因此，无透镜成像这种一体化方法，易于在低成本和紧凑型的装置中实现，以帮助噬菌体治疗诊断。
外文摘要：The growing number of drug-resistant bacterial infections worldwide is driving renewed interest in phage therapy. Based on the use of a personalized cocktail composed of highly specific bacterial viruses, this therapy relies on a range of tests on agar media to determine the most active phage on a given bacterial target (phage susceptibilitytesting), or to isolate new lytic phages from an environmental sample (enrichment of phage banks). However, these culture-based techniques are still solely interpreted through direct visual detection of plaques. The main objective of this work is to investigate computer-assisted methods in order to ease and accelerate diagnosis in phage therapy but also to study phage plaque growth kinetics. For this purpose, we designed a custom wide-field lensless imaging device, which allows continuous monitoring over a very large area sensor (3.3 cm(2)). Here we report bacterial susceptibility to Staphylococcus aureus phage in 3 hr and estimation of infectious titer in 8 hr 20 min. These are much shorter time-to-results than the 12 to 24 hours traditionally needed, since naked eye observation and counting of phage plaques is still the most widely used technique for susceptibility testing prior to phage therapy. Moreover, the continuous monitoring of the samples enables the study of plaque growth kinetics, which enables a deeper understanding of the interaction between phage and bacteria. Finally, thanks to the 4.3 mu m resolution, we detect phage-resistant bacterial microcolonies of Klebsiella pneumoniae inside the boundaries of phage plaques and thus show that our prototype is also a suitable device to track phage resistance. Lensless imaging is therefore an all-in-one method that could easily be implemented in cost-effective and compact devices in phage laboratories to help with phage therapy diagnosis.
作者：Perlemoine, P；Marcoux, PR；Picard, E；Hadji, E；Zelsmann, M；Mugnier, G；Marchet, A；Resch, G；O'Connell, L；Lacot, E
作者单位：Univ Grenoble Alpes；Univ Lausanne
期刊名称：PLOS ONE
期刊影响因子：2.74
出版年份：2021
出版刊次：3
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