Brandt, Christian, Zander, Esther, Pfeifer, Yvonne, Braun, Sascha D., Ehricht, Ralf, Makarewicz, Oliwia and Pletz, Mathias W. (2016). Development of a rapid diagnostic assay based on magnetic bead purification of OXA-beta-lactamase mRNA. Future Microbiol., 11 (5). S. 617 - 630. LONDON: FUTURE MEDICINE LTD. ISSN 1746-0921

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Abstract

Aim: To countermeasure the global spread of beta-lactamases, we developed a rapid molecular test for the highly variable OXA-beta-lactamases that allows minimizing the time to effective treatment. Methods: OXA-mRNA was specifically enriched from total RNA using group-specific biotinylated DNA probes and streptavidin-coated magnetic beads. Phylogenetic OXA groups were distinguished by PCR product size. Results: This mRNA fishing method is highly sensitive, yielding specific results from as little as 1 ng total RNA. It enables discrimination of OXA-extended substrate spectrum beta-lactamases and carbapenemases and the semi-quantitative detection of highly expressed ISAba1-controlled variants. Conclusion: Targeting mRNA with specific probes on magnetic beads will allow for adaptation to automated systems, such as point-of-care diagnostics.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Brandt, ChristianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Zander, EstherUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Pfeifer, YvonneUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Braun, Sascha D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ehricht, RalfUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Makarewicz, OliwiaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Pletz, Mathias W.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-289945
DOI: 10.2217/fmb.16.13
Journal or Publication Title: Future Microbiol.
Volume: 11
Number: 5
Page Range: S. 617 - 630
Date: 2016
Publisher: FUTURE MEDICINE LTD
Place of Publication: LONDON
ISSN: 1746-0921
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
KLEBSIELLA-PNEUMONIAE; ACINETOBACTER-BAUMANNII; ESCHERICHIA-COLI; GLOBAL SPREAD; INFECTIONS; RESISTANCE; ENTEROBACTERIACEAE; MORTALITY; IMPACT; CARBAPENEMSMultiple languages
MicrobiologyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/28994

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