MethodsX (Jan 2021)
Categorizing microbial growth inhibition through quantification of 16S rRNA growth marker with stripwells covering a spectrum of antimicrobial conditions
Abstract
Culture-based microdilution and disk diffusion tests are two commonly used reference methods for determining the susceptibility of causative bacteria to antibiotics. However, these methods are slow and laborious. Automated antimicrobial susceptibility test (AST) instruments are extensively used in clinical microbiology labs, replacing manual methods to perform gold standard microdilution or disk diffusion methods. These automated instruments require the use of isolated bacteria grown in pure culture against a fixed antimicrobial panel, and the susceptibility tests are based on measuring bacterial growth or turbidity changes over a range of pre-determined antimicrobial conditions. As a result, these automated technologies remain inherently inflexible to frequent adjustment of minimum inhibitory concentrations published by the Clinical and Laboratory Standards Institute and are limited by the detection methods that consumables were designed for. Here, we present a stripwell that is compatible with the 96-well format of most lab automation systems to provide a streamlined workflow to inoculate microorganisms for a customized or routine AST. The main goal of this method of stripwell preparation with various antibiotic conditions is to enable the utility of lab automation for phenotypic antibiotic response assays to address the reproducibility issues due to manual operation.• A standardized and scalable solution from inoculation to antimicrobial incubation• Microplates in stripwell format offer the advantage of greater flexibility in clinical microbiology and diagnostics• Customized antimicrobials and dilution ranges tailored to unique specifications for research and development
