Fungal-Grade Reagents and Materials for Molecular Analysis

Detail:

Series: Methods in Molecular Biology | Volume: 1508 | Pub. Date: Oct-08-2016 | Page Range: 141-150 | DOI: 10.1007/978-1-4939-6515-1_6

Year prepared: 2016

Fungal DNA is present at very low loads in clinical specimens. Molecular detection by amplification assays generally is a challenge because of a potentially multiple input of contaminating DNA from exogenous sources. Besides airborne, handling and cross-contamination, materials and reagents used in the molecular laboratory can contain microbial DNA which is a long underestimated potential source of false positive results. In this contribution decontamination procedures of materials and reagents and the selection of certified microbial DNA-free components for sample collection, DNA extraction, and PCR amplification are discussed with respect to the aim of building up a reliable molecular system for the diagnosis of fungal organisms at the limit of detection.

Laboratory Protocols

Select from:Additional methods & referenceAflatoxinsAssaying antifungal levelsAssaying mycotoxinsAztec Labs Diagnostic testing ProtocolsBiochemical CalculatorsBiofilmBiotypingCell Component ExtractionClinical BreakpointsCollection of Clinical MaterialCosmidsDetection in clinical samplesDNA extractionEnzyme synthesisFungal Molecular BiologyGeneral Molecular BiologyGeneticsGrowth and StorageHistologyMethods for DNA sequencingMicroscopyModel systems: in vitroModel systems: in vivoMore protocol collectionsPCR  -RT-qPCRPhagocytosisPlanar liquid chromatographyPost Antifungal Effects (PAFE)Preparation of amphotericin pasteProtein methodsProtocol overviewsQuality control in mycologySampling Aspergillus sporesSecondary metabolitesSpecies/Strain identification  -In situ hydridizationStandard Operating Procedures:National Aspergillosis CentreSusceptibility testingVeterinaryVideo Protocols at the FGSCWhole blood fractionationwhole-cell screening