Details

Autor(en) / Beteiligte

• Williamson, Charles H D
• Vazquez, Adam J
• Nunnally, Amalee E
• Kyger, Kristen
• Fofanov, Viacheslav Y
• Furstenau, Tara N
• Hornstra, Heidie M
• Terriquez, Joel
• Keim, Paul
• Sahl, Jason W
• Manning, Shannon D.

Titel

ColiSeq: a multiplex amplicon assay that provides strain level resolution of Escherichia coli directly from clinical specimens

Ist Teil von

• Microbiology spectrum, 2024-04, Vol.12 (6), p.e0413923-e0413923

Ort / Verlag

United States: American Society for Microbiology

Erscheinungsjahr

2024

Link zum Volltext

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• is a diverse pathogen, causing a range of disease in humans, from self-limiting diarrhea to urinary tract infections (UTIs). Uropathogenic (UPEC) is the most frequently observed uropathogen in UTIs, a common disease in high-income countries, incurring billions of dollars yearly in treatment costs. Although is easily grown and identified in the clinical laboratory, genotyping the pathogen is more complicated, yet critical for reducing the incidence of disease. These goals can be achieved through whole-genome sequencing of isolates, but this approach is relatively slow and typically requires culturing the pathogen in the laboratory. To genotype rapidly and inexpensively directly from clinical samples, including but not limited to urine, we developed and validated a multiplex amplicon sequencing assay, called ColiSeq. The assay consists of targets designed for species confirmation, high resolution genotyping, and mixture deconvolution. To demonstrate its utility, we screened the ColiSeq assay against 230 clinical urine samples collected from a hospital system in Flagstaff, Arizona, USA. A limit of detection analysis demonstrated the ability of ColiSeq to identify at a concentration of ~2 genomic equivalent (GEs)/mL and to generate high-resolution genotyping at a concentration of 1 × 10 GEs/mL. The results of this study suggest that ColiSeq could be a valuable method to understand the source of UPEC strains and guide infection mitigation efforts. As sequence-based diagnostics become accepted in the clinical laboratory, workflows such as ColiSeq will provide actionable information to improve patient outcomes.IMPORTANCEUrinary tract infections (UTIs), caused primarily by , create an enormous health care burden in the United States and other high-income countries. The early detection of from clinical samples, including urine, is important to target therapy and prevent further patient complications. Additionally, understanding the source of exposure will help with future mitigation efforts. In this study, we developed, tested, and validated an amplicon sequencing assay focused on direct detection of from urine. The resulting sequence data were demonstrated to provide strain level resolution of the pathogen, not only confirming the presence of , which can focus treatment efforts, but also providing data needed for source attribution and contact tracing. This assay will generate inexpensive, rapid, and reproducible data that can be deployed by public health agencies to track, diagnose, and potentially mitigate future UTIs caused by .