Rapid Antibiotic Susceptibility Testing of Uropathogenic E. coli by Tracking Submicron Scale Motion of Single Bacterial Cells

Abstract

To combat antibiotic resistance, a rapid antibiotic susceptibility testing (AST) technology that can identify resistant infections at disease onset is required. Current clinical AST technologies take 1-3 days, which is often too slow for accurate treatment. Here we demonstrate a rapid AST method by tracking sub-μm scale bacterial motion with an optical imaging and tracking technique. We apply the method to clinically relevant bacterial pathogens, Escherichia coli O157: H7 and uropathogenic E. coli (UPEC) loosely tethered to a glass surface. By analyzing dose-dependent sub-μm motion changes in a population of bacterial cells, we obtain the minimum bactericidal concentration within 2 h using human urine samples spiked with UPEC. We validate the AST method using the standard culture-based AST methods. In addition to population studies, the method allows single cell analysis, which can identify subpopulations of resistance strains within a sample.

Microfluidics as an Emerging Platform for Tackling Antimicrobial Resistance (AMR): A Review

“…Single cell imaging of the bacteria treated with antibiotics can also provide valuable information about the resistivity of the microbes [26][27][28] such as Yoshimi and co-workers [26] developed drug susceptibility testing microfluidic (DSTM) device to perform AST of Pseudomonas aeruginosa within 3 hours. The device consisted of five sets of microchannels with one inlet and four narrowly fabricated channels for the microscopic view.…”

“…Different types of drugs reacted differently with the bacteria such as susceptible cells elongated upon incubation with ciprofloxacin, ceftazidime, meropenem and piperacillin while amikacin affected the cell shape minorly. In another bacterial imaging technique, Syal et al [27] developed an imaging and tracking technology to identify sub-µm bacterial cell motions (Fig.1d). E coli O157:H7 and uropathogenic E coli (UPEC) were placed on a glass surface, and their sub-µm movements were tracked upon treatment with antibiotics.…”

A WS2-gold nanoparticle heterostructure-based novel SERS platform for the rapid identification of antibiotic-resistant pathogens

“…26 As a result, there is a huge urgency in nding rapid and costeffective assays which will have the capability for rapid and accurate determination of the appropriate antibiotic for an infection in a timely manner. [31][32][33][34][35][36][37][38][39][40][41][42] Driven by this need, the current manuscript demonstrates that 0D-2D heterostructurebased SERS is capable of rapid identication of drug resistant bacteria by monitoring structural changes in the pathogen's cell wall during antibiotic treatment. Reported data show that the SERS can be used for the identication of multidrug resistant Salmonella DT104 and a normal strain Salmonella Typhi antimicrobial susceptibility test using Augmentin antibiotics, even at the concentration of 100 CFU mL À1 experimental data reported here indicates that heterostructure-based SERS has the capability of antimicrobial susceptibility testing in less than 2 hours, whereas the "gold standard" broth microdilution (BMD) test takes 16 to 20 h in clinics until the number of bacteria reaches above 10 5 CFU mL À1 .…”

A Rapid Unraveling of the Activity and Antibiotic Susceptibility of Mycobacteria

“…Many options are emerging to achieve a rapid, accurate, and cost-effective pathogen characterization of bacterial responses to drugs, ranging from molecular to rapid phenotypic techniques to plasmonic single-cell assays (8)(9)(10)(11). The conventional AST molecular techniques rely mainly on the determination of the genetic fingerprint associated with resistance to a specific antibiotic, including real-time PCR (RT-PCR), DNA microarrays, next-generation sequencing (NGS), cell lysis-based approaches, wholegenome sequencing, and MALDI-TOF MS (12)(13)(14)(15)(16).…”

“…Among the alternatives to conventional microbiological assays, small and extremely sensitive nanomechanical oscillators (19) stand out as very promising candidates (8,9,20,21). At first, such devices were employed in atomic force microscopes (AFMs) to study dynamic behavior in cells (22) or proteins (23,24).…”