Digital Quantification of DNA Replication and Chromosome Segregation Enables Determination of Antimicrobial Susceptibility after only 15 Minutes of Antibiotic Exposure

Rapid antimicrobial susceptibility testing (AST) would decrease misuse and overuse of antibiotics. The “holy grail” of AST is a phenotype‐based test that can be performed within a doctor visit. Such a test requires the ability to determine a pathogen's susceptibility after only a short antibiot...

Full description

Saved in:
Bibliographic Details
Published in:Angewandte Chemie International Edition 2016-08, Vol.55 (33), p.9557-9561
Main Authors: Schoepp, Nathan G., Khorosheva, Eugenia M., Schlappi, Travis S., Curtis, Matthew S., Humphries, Romney M., Hindler, Janet A., Ismagilov, Rustem F.
Format: Article
Language:English
Subjects:
analytical methods
Anti-Bacterial Agents - pharmacology
antibiotics
Chromosome Segregation - drug effects
DNA replication
DNA Replication - drug effects
DNA, Bacterial - drug effects
DNA, Bacterial - genetics
Polymerase Chain Reaction
susceptibility
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Rapid antimicrobial susceptibility testing (AST) would decrease misuse and overuse of antibiotics. The “holy grail” of AST is a phenotype‐based test that can be performed within a doctor visit. Such a test requires the ability to determine a pathogen's susceptibility after only a short antibiotic exposure. Herein, digital PCR (dPCR) was employed to test whether measuring DNA replication of the target pathogen through digital single‐molecule counting would shorten the required time of antibiotic exposure. Partitioning bacterial chromosomal DNA into many small volumes during dPCR enabled AST results after short exposure times by 1) precise quantification and 2) a measurement of how antibiotics affect the states of macromolecular assembly of bacterial chromosomes. This digital AST (dAST) determined susceptibility of clinical isolates from urinary tract infections (UTIs) after 15 min of exposure for all four antibiotic classes relevant to UTIs. This work lays the foundation to develop a rapid, point‐of‐care AST and strengthen global antibiotic stewardship. Single‐molecule counting was used to detect subtle changes in the amount and the assembly state of bacterial chromosomes after a short exposure of live bacteria to antibiotics. The method employed digital PCR to determine the susceptibility of E. coli clinical isolates from urinary tract infections to four different antibiotics.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.201602763