A single-cell drug efflux assay in bacteria by using a directly accessible femtoliter droplet array

作者:Iino Ryota*; Hayama Kohei; Amezawa Hiromi; Sakakihara Shouichi; Kim Soo Hyeon; Matsumono Yoshimi; Nishino Kunihiko; Yamaguchi Akihito; Noji Hiroyuki
来源:Lab on a Chip, 2012, 12(20): 3923-3929.
DOI:10.1039/c2lc40394c

摘要

Active efflux of drugs, such as antibiotics, from a cell is one of the major mechanisms that cause multi-drug resistance in bacteria. Here we report a method to assess drug efflux activity in individual Escherichia coli cells enclosed and isolated in a directly accessible femtoliter droplet array with a fluorogenic compound. The inhibitory effect of a chemical compound on an exogenously expressed efflux pump system from pathogenic bacteria was easily detected at the single-cell level. We also present a proof-of-principle experiment to screen for the gene encoding a drug efflux pump by collecting individual droplets containing single cells in which the drug efflux activity was restored after introduction of the exogenous gene from pathogenic bacteria. Our approach will be a useful tool to screen novel pump inhibitors and efflux pump genes, and to overcome infectious diseases caused by multi-drug-resistant bacteria.

  • 出版日期2012