Utility of Real-Time PCR for Detection of Exserohilum rostratum in Body and Tissue Fluids during the Multistate Outbreak of Fungal Meningitis and Other Infections

dc.contributor.authorGade, Lalithaen
dc.contributor.authorGrgurich, Dale E.en
dc.contributor.authorKerkering, Thomas M.en
dc.contributor.authorBrandt, Mary E.en
dc.contributor.authorLitvintseva, Anastasia P.en
dc.contributor.departmentVirginia Tech Carilion School of Medicineen
dc.contributor.editorDiekema, D. J.en
dc.date.accessed2015-11-28en
dc.date.accessioned2015-11-29T04:13:28Zen
dc.date.available2015-11-29T04:13:28Zen
dc.date.issued2015en
dc.description.abstractExserohilum rostratum was the major cause of the multistate outbreak of fungal meningitis linked to contaminated injections of methylprednisolone acetate produced by the New England Compounding Center. Previously, we developed a fungal DNA extraction procedure and broad-range and E. rostratum-specific PCR assays and confirmed the presence of fungal DNA in 28% of the case patients. Here, we report the development and validation of a TaqMan real-time PCR assay for the detection of E. rostratum in body fluids, which we used to confirm infections in 57 additional case patients, bringing the total number of case patients with PCR results positive for E. rostratum to 171 (37% of the 461 case patients with available specimens). Compared to fungal culture and the previous PCR assays, this real-time PCR assay was more sensitive. Of the 139 identical specimens from case patients tested by all three methods, 19 (14%) were positive by culture, 41 (29%) were positive by the conventional PCR assay, and 65 (47%) were positive by the real-time PCR assay. We also compared the utility of the real-time PCR assay with that of the previously described beta-D-glucan (BDG) detection assay for monitoring response to treatment in case patients with serially collected CSF. Only the incident CSF specimens from most of the case patients were positive by real-time PCR, while most of the subsequently collected specimens were negative, confirming our previous observations that the BDG assay was more appropriate than the real-time PCR assay for monitoring the response to treatment. Our results also demonstrate that the real-time PCR assay is extremely susceptible to contamination and its results should be used only in conjunction with clinical and epidemiological data.en
dc.format.mimetypeapplication/pdfen
dc.identifier.citationGade, Lalitha, et al. (2015). Utility of Real-Time PCR for Detection of Exserohilum rostratum in Body and Tissue Fluids during the Multistate Outbreak of Fungal Meningitis and Other Infections. Journal of Clinical Microbiology, 53(2), 618-625. doi:10.1128/jcm.02443-14en
dc.identifier.doihttps://doi.org/10.1128/jcm.02443-14en
dc.identifier.issn0095-1137en
dc.identifier.urihttp://hdl.handle.net/10919/64228en
dc.identifier.urlhttp://jcm.asm.org/content/53/2/618en
dc.language.isoen_USen
dc.publisherAmerican Society for Microbiologyen
dc.rightsIn Copyright (InC)en
dc.rightsThis Item is protected by copyright and/or related rights. Some uses of this Item may be deemed fair and permitted by law even without permission from the rights holder(s). For other uses you need to obtain permission from the rights holder(s).en
dc.rights.holderAmerican Society for Microbiologyen
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.titleUtility of Real-Time PCR for Detection of Exserohilum rostratum in Body and Tissue Fluids during the Multistate Outbreak of Fungal Meningitis and Other Infectionsen
dc.title.serialJournal of Clinical Microbiologyen
dc.typeArticle - Refereeden
dc.typeDataseten
dc.type.dcmitypeTexten
dc.type.dcmitypeDataseten

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