Department of Biomedical Engineering, University of Connecticut, Storrs, CT 06269, USA.
Department of Chemical and Biomolecular Engineering, University of Connecticut, Storrs, CT 06269, USA.
Molecules. 2019 Jul 3;24(13):2441. doi: 10.3390/molecules24132441.
Since conventional culture-based antibiotic susceptibility testing (AST) methods are too time-consuming (typically 24-72 h), rapid AST is urgently needed for preventing the increasing emergence and spread of antibiotic resistant infections. Although several phenotypic antibiotic resistance sensing modalities are able to reduce the AST time to a few hours or less, concerning the biological heterogeneity, their accuracy or limit of detection are limited by low throughput. Here, we present a rapid AST method based on whole slide imaging (WSI)-enabled high-throughput sensing antibiotic resistance at single-bacterium level. The time for determining the minimum inhibitory concentration (MIC) was theoretically shortest, which ensures that the growth of each individual cell present in a large population is inhibited. As a demonstration, our technique was able to sense the growth of at least several thousand bacteria at single-cell level. Reliable MIC of against gentamicin was obtained within 1 h, while the gold standard broth dilution method required at least 16 h for the same result. In addition, the application of our method prevails over other imaging-based AST approaches in allowing rapid and accurate determination of antibiotic susceptibility for phenotypically heterogeneous samples, in which the number of antibiotic resistant cells was negligible compared to that of the susceptible cells. Hence, our method shows great promise for both rapid AST determination and point-of-care testing of complex clinical bacteria isolates.
由于传统的基于培养的抗生素药敏试验（AST）方法过于耗时（通常需要 24-72 小时），因此迫切需要快速 AST 来防止抗生素耐药性感染的不断出现和传播。尽管几种表型抗生素耐药性感应模式能够将 AST 时间缩短到几个小时或更短，但由于生物异质性，其准确性或检测限受到低通量的限制。在这里，我们提出了一种基于全玻片成像（WSI）的快速 AST 方法，可在单细胞水平上实现高通量感应抗生素耐药性。确定最小抑菌浓度（MIC）的时间最短，可确保大量存在的每个细胞的生长都受到抑制。作为演示，我们的技术能够在单细胞水平上检测至少数千个细菌的生长。在 1 小时内可靠地获得了对庆大霉素的 MIC 值，而金标准肉汤稀释法则需要至少 16 小时才能得到相同的结果。此外，与其他基于成像的 AST 方法相比，我们的方法在允许快速准确地确定表型异质样本中的抗生素敏感性方面具有优势，其中耐药细胞的数量与敏感细胞相比可以忽略不计。因此，我们的方法在快速 AST 测定和复杂临床细菌分离物的即时检测方面具有广阔的应用前景。
