Document Detail


Direct detection and genotyping of Klebsiella pneumoniae carbapenemases from urine by use of a new DNA microarray test.
MedLine Citation:
PMID:  23035190     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Klebsiella pneumoniae carbapenemases (KPCs) are considered a serious threat to antibiotic therapy, as they confer resistance to carbapenems, which are used to treat extended-spectrum beta-lactamase (ESBL)-producing bacteria. Here, we describe the development and evaluation of a DNA microarray for the detection and genotyping of KPC genes (bla(KPC)) within a 5-h period. To test the whole assay procedure (DNA extraction plus a DNA microarray assay) directly from clinical specimens, we compared two commercial DNA extraction kits (the QIAprep Spin miniprep kit [Qiagen] and the urine bacterial DNA isolation kit [Norgen]) for the direct DNA extraction from urine samples (dilution series spiked in human urine). Reliable single nucleotide polymorphism (SNP) typing was demonstrated using 1 × 10(5) CFU/ml urine for Escherichia coli (Qiagen and Norgen) and 80 CFU/ml urine, on average, for K. pneumoniae (Norgen). This study presents, for the first time, the combination of a new KPC microarray with commercial sample preparation for detecting and genotyping microbial pathogens directly from clinical specimens; this paves the way toward tests providing epidemiological and diagnostic data, enabling better antimicrobial stewardship.
Authors:
Harald Peter; Kathrine Berggrav; Peter Thomas; Yvonne Pfeifer; Wolfgang Witte; Kate Templeton; Till T Bachmann
Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-10-03
Journal Detail:
Title:  Journal of clinical microbiology     Volume:  50     ISSN:  1098-660X     ISO Abbreviation:  J. Clin. Microbiol.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-11-19     Completed Date:  2013-04-17     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  7505564     Medline TA:  J Clin Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3990-8     Citation Subset:  IM    
Affiliation:
Division of Pathway Medicine, University of Edinburgh, Medical School, Edinburgh, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics*
Enterobacteriaceae Infections / microbiology*
Escherichia coli
Humans
Klebsiella pneumoniae / enzymology*,  genetics,  isolation & purification
Oligonucleotide Array Sequence Analysis / methods*
Urine / microbiology*
beta-Lactamases / genetics*
Chemical
Reg. No./Substance:
0/Bacterial Proteins; EC 3.5.2.6/beta-Lactamases; EC 3.5.2.6/carbapenemase
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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