“Antibiotic inhibition of bacteria growth in droplets reveals heteroresistance pattern at the single cell level”

Abstract

Heteroresistance is a phenomenon where isogenic bacteria population exhibits a diverse antibiotic resistance pattern at sub-population or single cell level. The sub-populations with higher resistance can remain undetected with conventional diagnostics which makes them subsequently harder to treat. Such surviving phenotypically heterogeneous sub-populations are also a potential hotbed for novel mutations, thus increasing the resistance permanently in bacteria. Droplet microfluidics gives tools for high-throughput analysis of bacteria and their response to antibiotics at single cell level, which is difficult to obtain with traditional agar plate technologies. In here we show for the first time the precise digital quantification of drug resistance profile in isogenic population at single cell level. We also see that the inhibiting amount of drug per bacteria remains quite stable regardless of bacteria density. Interestingly, the bacteria clump together preferably near these sub-inhibitory conditions. The technology and findings we describe here provide novel quantitative insight into the heteroresistance which is a key step in understanding the pathways leading to drug resistance. This knowledge is crucial in the context of global drug resistance threat as it can help us to find tools to prevent further escalation of drug resistance.