Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms

Sci Rep. 2021 Aug 16;11(1):16572. doi: 10.1038/s41598-021-96148-3.

Abstract

Routinely used typing methods including MLST, rep-PCR and whole genome sequencing (WGS) are time-consuming, costly, and often low throughput. Here, we describe a novel mini-MLST scheme for Eschericha coli as an alternative method for rapid genotyping. Using the proposed mini-MLST scheme, 10,946 existing STs were converted into 1,038 Melting Types (MelTs). To validate the new mini-MLST scheme, in silico analysis was performed on 73,704 strains retrieved from EnteroBase resulting in discriminatory power D = 0.9465 (CI 95% 0.9726-0.9736) for mini-MLST and D = 0.9731 (CI 95% 0.9726-0.9736) for MLST. Moreover, validation on clinical isolates was conducted with a significant concordance between MLST, rep-PCR and WGS. To conclude, the great portability, efficient processing, cost-effectiveness, and high throughput of mini-MLST represents immense benefits, even when accompanied with a slightly lower discriminatory power than other typing methods. This study proved mini-MLST is an ideal method to screen and subgroup large sets of isolates and/or quick strain typing during outbreaks. In addition, our results clearly showed its suitability for prospective surveillance monitoring of emergent and high-risk E. coli clones'.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Typing Techniques*
  • Base Composition
  • Computer Simulation
  • Czech Republic / epidemiology
  • DNA Primers
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics*
  • Disease Outbreaks
  • Escherichia coli / classification
  • Escherichia coli / genetics*
  • Escherichia coli / isolation & purification
  • Escherichia coli Infections / microbiology
  • Genes, Bacterial*
  • Genome, Bacterial
  • Genotyping Techniques*
  • Multilocus Sequence Typing / methods*
  • Nucleic Acid Denaturation
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single Nucleotide*
  • Population Surveillance
  • Repetitive Sequences, Nucleic Acid
  • Whole Genome Sequencing

Substances

  • DNA Primers
  • DNA, Bacterial