title: Increased sensitivity of bacterial detection in cerebrospinal fluid by fluorescent staining on low-fluorescence membrane filters.
abstract: A membrane-filter-based, fluorescent Gram stain method for bacterial detection in cerebrospinal fluid samples was developed and evaluated as a rapid, sensitive alternative to standard Gram stain protocols. A recently developed, modified version of the aluminium oxide membrane Anopore with low-fluorescence optical properties showed superior performance in this application. Other aspects of the fluorescent Gram stain system that were evaluated include membrane filter selection, strategies to reduce fluorescence fading and the effect of patient blood cells on bacterial detection in the fluorescently stained cerebrospinal fluid samples. The combination of the membrane filter's bacteria-concentrating ability and absolute retention along with high-contrast, fluorescent Gram discriminating dyes enabled rapid bacterial detection and Gram discrimination, with a 1-1.5 order of magnitude increase in the bacterial concentration limit of detection.
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Jacob D
authors.initials: JD
authors.lastname: Durtschi
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Erali
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: L Kathryn
authors.initials: LK
authors.lastname: Bromley
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Mark G
authors.initials: MG
authors.lastname: Herrmann
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Cathy A
authors.initials: CA
authors.lastname: Petti
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Roger E
authors.initials: RE
authors.lastname: Smith
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology1 and Department of Pathology2, University of Utah, Salt Lake City, UT 84108, USA.
authors.forename: Karl V
authors.initials: KV
authors.lastname: Voelkerding
mesh_terms: D001431:Bacteriological Techniques; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000302:isolation & purification; D005374:Filtration / Q000295:instrumentation / Q000379:methods; D005453:Fluorescence; D005840:Gentian Violet; D006801:Humans; D008851:Micropore Filters; D008856:Microscopy, Fluorescence / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010619:Phenazines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D013211:Staphylococcus aureus / Q000302:isolation & purification
chemical_list: C035863:Gram's stain; D009930:Organic Chemicals; D010619:Phenazines; C103389:SYTO 9; D005840:Gentian Violet