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Evaluation of loop-mediated isothermal amplification for the rapid identification of bacteria and resistance determinants in positive blood cultures

Rödel, J. ; Bohnert, J. A. ; Stoll, S. ; Wassill, L. ; Edel, B. ; Karrasch, M. ; Löffler, B. ; Pfister, W.

European journal of clinical microbiology & infectious diseases, 2017-06, Vol.36 (6), p.1033-1040 [Periódico revisado por pares]

Berlin/Heidelberg: Springer Berlin Heidelberg

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Título:
Evaluation of loop-mediated isothermal amplification for the rapid identification of bacteria and resistance determinants in positive blood cultures
Autor: Rödel, J. ; Bohnert, J. A. ; Stoll, S. ; Wassill, L. ; Edel, B. ; Karrasch, M. ; Löffler, B. ; Pfister, W.
Assuntos: Antibiotics ; Assaying ; Bacteremia - diagnosis ; Bacteria ; Bacteria - drug effects ; Bacteria - genetics ; Bacteria - isolation & purification ; Biomedical and Life Sciences ; Biomedicine ; Blood ; Blood Culture ; Diagnosis ; Diagnostic systems ; Drug resistance ; Drug Resistance, Bacterial ; Fluorescence ; Genes ; Genotyping Techniques - methods ; Humans ; Internal Medicine ; Medical Microbiology ; Microbial Sensitivity Tests - methods ; Molecular Diagnostic Techniques - methods ; Nucleic Acid Amplification Techniques - methods ; Original Article ; Sample preparation ; Sensitivity and Specificity ; Sepsis ; Time Factors
É parte de: European journal of clinical microbiology & infectious diseases, 2017-06, Vol.36 (6), p.1033-1040
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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Descrição: The use of molecular assays to rapidly identify pathogens and resistance genes directly from positive blood cultures (BCs) contribute to shortening the time required for the diagnosis of bloodstream infections. In this work, loop-mediated isothermal amplification (LAMP) assays have been examined for their potential use in BC diagnosis. Three different assays were applied. The commercially available eazyplex® MRSA test detects Staphylococcus aureus , S. epidermidis , mecA , and mecC . Two in-house assays [Gram-positive (GP) and Gram-negative (GN)] have been developed for the detection of streptococci, enterococci, vanA , vanB , Pseudomonas spp., Enterobacteriaceae, and the bla CTX-M family. A total of 370 positive BCs were analyzed. LAMP test results were obtained within 30 min, including sample preparation. Amplification was measured by real-time fluorescence detection. The threshold time for fluorescence intensity values ranged from 6.25 to 13.75 min. The specificity and sensitivity of the assays varied depending on the target. Overall, from 87.7% of BCs, true-positive results were obtained, compared to routine standard diagnosis. Twenty-one tests were true-negative because of the lack of an appropriate target (5.7%). The concordance of positive test results for resistance genes with subsequent antibiotic susceptibility testing was 100%. From 15 BC bottles with mixed cultures, eazyplex® assays produced correct results in 73% of the cases. This study shows that LAMP assays are fast and cost-saving tools for rapid BC testing in order to expedite the diagnostic report and improve the antibiotic stewardship for sepsis patients.
Editor: Berlin/Heidelberg: Springer Berlin Heidelberg
Idioma: Inglês

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Evaluation of loop-mediated isothermal amplification for the rapid identification of bacteria and resistance determinants in positive blood cultures

Rödel, J. ; Bohnert, J. A. ; Stoll, S. ; Wassill, L. ; Edel, B. ; Karrasch, M. ; Löffler, B. ; Pfister, W.

Berlin/Heidelberg: Springer Berlin Heidelberg

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