Predicting antimicrobial susceptibilities for Escherichia coli and Klebsiella pneumoniae isolates using whole genomic sequence data

Whole-genome sequencing potentially represents a single, rapid and cost-effective approach to defining resistance mechanisms and predicting phenotype, and strain type, for both clinical and epidemiological purposes. This retrospective study aimed to determine the efficacy of whole genome-based antim...

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Bibliographic Details
Published in:	Journal of antimicrobial chemotherapy Vol. 68; no. 10; pp. 2234 - 2244
Main Authors:	Stoesser, N, Batty, E M, Eyre, D W, Morgan, M, Wyllie, D H, Del Ojo Elias, C, Johnson, J R, Walker, A S, Peto, T E A, Crook, D W
Format:	Journal Article
Language:	English
Published:	England Oxford Publishing Limited (England) 01-10-2013 Oxford University Press
Subjects:	Anti-Bacterial Agents - pharmacology Bacteremia - microbiology Chromosomes DNA, Bacterial - genetics Drug resistance Drug Resistance, Bacterial E coli Epidemiology Escherichia coli Escherichia coli - drug effects Escherichia coli - genetics Escherichia coli - isolation & purification Genome, Bacterial Genomics Genotype & phenotype Humans Klebsiella pneumoniae Klebsiella pneumoniae - drug effects Klebsiella pneumoniae - genetics Klebsiella pneumoniae - isolation & purification Microbial Sensitivity Tests - methods Mutation Original Research Sensitivity and Specificity Sequence Analysis, DNA United Kingdom United Kingdom United Kingdom > UK antibiotic phenotype Gram-negative
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Summary:	Whole-genome sequencing potentially represents a single, rapid and cost-effective approach to defining resistance mechanisms and predicting phenotype, and strain type, for both clinical and epidemiological purposes. This retrospective study aimed to determine the efficacy of whole genome-based antimicrobial resistance prediction in clinical isolates of Escherichia coli and Klebsiella pneumoniae. Seventy-four E. coli and 69 K. pneumoniae bacteraemia isolates from Oxfordshire, UK, were sequenced (Illumina HiSeq 2000). Resistance phenotypes were predicted from genomic sequences using BLASTn-based comparisons of de novo-assembled contigs with a study database of >100 known resistance-associated loci, including plasmid-associated and chromosomal genes. Predictions were made for seven commonly used antimicrobials: amoxicillin, co-amoxiclav, ceftriaxone, ceftazidime, ciprofloxacin, gentamicin and meropenem. Comparisons were made with phenotypic results obtained in duplicate by broth dilution (BD Phoenix). Discrepancies, either between duplicate BD Phoenix results or between genotype and phenotype, were resolved with gradient diffusion analyses. A wide variety of antimicrobial resistance genes were identified, including blaCTX-M, blaLEN, blaOKP, blaOXA, blaSHV, blaTEM, aac(3')-Ia, aac-(3')-IId, aac-(3')-IIe, aac(6')-Ib-cr, aadA1a, aadA4, aadA5, aadA16, aph(6')-Id, aph(3')-Ia, qnrB and qnrS, as well as resistance-associated mutations in chromosomal gyrA and parC genes. The sensitivity of genome-based resistance prediction across all antibiotics for both species was 0.96 (95% CI: 0.94-0.98) and the specificity was 0.97 (95% CI: 0.95-0.98). Very major and major error rates were 1.2% and 2.1%, respectively. Our method was as sensitive and specific as routinely deployed phenotypic methods. Validation against larger datasets and formal assessments of cost and turnaround time in a routine laboratory setting are warranted.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Undefined-1 ObjectType-Feature-3 content type line 23 ObjectType-Feature-1
ISSN:	0305-7453 1460-2091
DOI:	10.1093/jac/dkt180