Generation of uniform emulsion droplets mixed with multiple components is one of the key issues in the field of lab in a drop. Traditionally, droplet microfluidic chips are often served as the prime choice while designing and fabricating microfluidic chips always rely on skilled technician and specialized equipment, severely restricting its wide accessibility. In this work, an alternative technique, called multichannel dynamic interfacial printing (MC-DIP), was proposed for multicomponent droplet generation. The MC-DIP device was designed modularly and could be set up manually without any microfabrication process, exhibiting full accessibility for freshmen after a brief training. This new technique owns advantages in the generation of droplets with predictable sizes and composites. Quantitative experiments of measuring minimum inhibitory concentration (MIC) value via mixing microbes and antibiotics into droplet were conducted to proving its application potential for lab in a drop. Further research on a clinical pathogenic strain revealed that this technique could be potentially applied in the clinical laboratory for antibiotic susceptibility testing.
The rapid spread of carbapenemase-producing Klebsiella pneumoniae (cpKP) poses serious threats to public health; however, the underlying genetic basis for its dissemination is still unknown. We conducted a comprehensive genomic epidemiology analysis on 420 cpKP isolates collected from 70 hospitals in 24 provinces/autonomous regions/municipalities of China during 2009–2017 by short-/long-read sequencing. The results showed that most cpKP isolates were categorized into clonal group 258 (CG258), in which ST11 was the dominant clone. Phylogenetic analysis revealed three major clades including the top one of Clade 3 for CG258 cpKP isolates. Additionally, carbapenemase gene analysis indicated that blaKPC was dominant in the cpKP isolates, and most blaKPC genes were located in five major incompatibility (Inc) groups of blaKPC-harboring plasmids. Importantly, three advantageous combinations of host–blaKPC-carrying plasmid (Clade 3.1+3.2–IncFIIpHN7A8, Clade 3.1+3.2–IncFIIpHN7A8:IncR, and Clade 3.3–IncFIIpHN7A8:IncpA1763-KPC) were identified to confer cpKP isolates the advantages in both genotypes (strong correlation/coevolution) and phenotypes (resistance/growth/competition) to facilitate the nationwide spread of ST11/CG258 cpKP. Intriguingly, Bayesian skyline analysis illustrated that the three advantageous combinations might be directly associated with the strong population expansion during 2007–2008 and subsequent maintenance of the population of ST11/CG258 cpKP after 2008. We then examined drug resistance profiles of these cpKP isolates and proposed combination treatment regimens for CG258/non-CG258 cpKP infections. Thus, the findings of our systematical analysis shed light on the molecular epidemiology and genetic basis for the dissemination of ST11/CG258 cpKP in China, and much emphasis should be given to the close monitoring of advantageous cpKP–plasmid combinations.
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