Rapid antibiotic susceptibility testing by tracking single cell growth in a microfluidic agarose channel system

Lab on a Chip ◽  
2013 ◽  
Vol 13 (2) ◽  
pp. 280-287 ◽  
Author(s):  
Jungil Choi ◽  
Yong-Gyun Jung ◽  
Jeewoo Kim ◽  
Sungbum Kim ◽  
Yushin Jung ◽  
...  
2021 ◽  
Author(s):  
Wenshuai Wu ◽  
Gaozhe Cai ◽  
Yang Liu

On-site single-cell antibiotic susceptibility testing (sc-AST) provides unprecedented technical potential to improve the treatment of bacterial infections and study heterogeneous resistance to antibiotics. Herein, we developed a portable and high-integrated 3D polydimethylsiloxane (PDMS) chip to perform fast and on-site bacteria quantification and sc-AST. The 3D arrangement of the chambers significantly improved the integration of reaction units (~10000/cm2) and widened the dynamic range to 5 orders of magnitude. A capillary valve-based flow distributor was adopted for flow equidistribution in 64 parallel channels and uniform sample loading in as short as 2 s. The degassed PDMS enabled this device to independently dispense the sample into 3D chamber array with almost 100% efficiency. The quantification of Escherichia coli (E. coli) strains with various activity was accomplished in 0.5-2 h, shortened by 20 h in comparison to the traditional plate counting. The functionality of our platform was demonstrated with several effective antibiotics by determining minimum inhibitory concentrations at single-cell level. Furthermore, we utilized the lyophilization of test reagents and needle-mediated reagents rehydration to realize one-step on-site sc-AST. The results indicate that the proposed sc-AST platform is portable, highly sensitive, fast, accurate and user-friendly, thus it has the potential to facilitate precise therapy in time and monitor the treatment. Meanwhile, it could serve as an approach for investigating the mechanisms of heteroresistance at single-cell resolution.


ACS Omega ◽  
2021 ◽  
Author(s):  
Armelle Novelli Rousseau ◽  
Nicolas Faure ◽  
Fabian Rol ◽  
Zohreh Sedaghat ◽  
Joël Le Galudec ◽  
...  

2020 ◽  
Vol 41 (S1) ◽  
pp. s42-s43
Author(s):  
Kimberley Sukhum ◽  
Candice Cass ◽  
Meghan Wallace ◽  
Caitlin Johnson ◽  
Steven Sax ◽  
...  

Background: Healthcare-associated infections caused by antibiotic-resistant organisms (AROs) are a major cause of significant morbidity and mortality. To create and optimize infection prevention strategies, it is crucial to delineate the role of the environment and clinical infections. Methods: Over a 14-month period, we collected environmental samples, patient feces, and patient bloodstream infection (BSI) isolates in a newly built bone marrow transplant (BMT) intensive care unit (ICU). Samples were collected from 13 high-touch areas in the patient room and 4 communal areas. Samples were collected from the old BMT ICU, in the new BMT ICU before patients moved in, and for 1 year after patients moved in. Selective microbiologic culture was used to isolate AROs, and whole-genome sequencing (WGS) was used to determine clonality. Antibiotic susceptibility testing was performed using Kirby-Bauer disk diffusion assays. Using linear mixed modeling, we compared ARO recovery across time and sample area. Results: AROs were collected and cultured from environmental samples, patient feces, and BSI isolates (Fig. 1a). AROs were found both before and after a patient entered the ICU (Fig. 1b). Sink drains had significantly more AROs recovered per sample than any other surface area (P < .001) (Fig. 1c). The most common ARO isolates were Pseudomonas aeruginosa and Stenotrophomonas maltophila (Fig. 1d). The new BMT ICU had fewer AROs recovered per sample than the old BMT ICU (P < .001) and no increase in AROs recovered over the first year of opening (P > .05). Furthermore, there was no difference before versus after patients moved into the hospital (P > .05). Antibiotic susceptibility testing reveal that P. aeruginosa isolates recovered from the old ICU were resistant to more antibiotics than isolates recovered from the new ICU (Fig. 2a). ANI and clonal analyses of P. aeruginosa revealed a large cluster of clonal isolates (34 of 76) (Fig. 2b). This clonal group included isolates found before patients moved into the BMT ICU and patient blood isolates. Furthermore, this clonal group was initially found in only 1 room in the BMT ICU, and over 26 weeks, it was found in sink drains in all 6 rooms sampled (Fig. 2b). Conclusions: AROs are present before patients move into a new BMT ICU, and sink drains act as a reservoir for AROs over time. Furthermore, sink-drain P. aeruginosa isolates are clonally related to isolates found in patient BSIs. Overall, these results provide insight into ARO transmission dynamics in the hospital environment.Funding: Research reported in this publication was supported by the Washington University Institute of Clinical and Translational Sciences grant UL1TR002345 from the National Center for Advancing Translational Sciences (NCATS) of the National Institutes of Health (NIH). The content is solely the responsibility of the authors and does not necessarily represent the official view of the NIH.Disclosures: None


1976 ◽  
Vol 10 (3) ◽  
pp. 436-440 ◽  
Author(s):  
M. B. Coyle ◽  
M. F. Lampe ◽  
C. L. Aitken ◽  
P. Feigl ◽  
J. C. Sherris

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