A major public health issue is the growing resistance of bacteria to antibiotics. An important part of the needed response is the discovery and development of new antimicrobial strategies. These require the screening of potential new drugs, typically accomplished using high-throughput screening (HTS). Traditionally, HTS is performed by examining one compound per well, but a more efficient strategy pools multiple compounds per well. In this work, we study several recently proposed pooling construction methods, as well as a variety of pooled high-throughput screening analysis methods, in order to provide guidance to practitioners on which methods to use. This is done in the context of an application of the methods to the search for new drugs to combat bacterial infection. We discuss both an extensive pilot study as well as a small screening campaign, and highlight both the successes and challenges of the pooling approach.

翻译：细菌对抗生素的耐药性日益增强已成为一项重大的公共卫生问题。应对这一挑战的关键环节在于发现和开发新型抗菌策略，这通常需要通过高通量筛选（HTS）对潜在新药进行筛选。传统的高通量筛选采用每孔检测单一化合物的方式，而将多个化合物混合于单孔中的池化策略则能显著提升筛选效率。本研究系统评估了近期提出的多种池化构建方法以及多样化的池化高通量筛选分析策略，旨在为实践者提供方法选择的指导。研究以寻找抗细菌感染新药为应用背景，通过开展大规模先导研究和小规模筛选实验，全面探讨了池化策略取得的成效与面临的挑战。