Real-Time Quantitative PCR, Pathogen Detection and MIQE

Year prepared: 2009

Nucleic acids are the ultimate biomarker and real-time PCR (qPCR) is firmly established as the method of choice for nucleic acid detection. Together, they allow the accurate, sensitive and specific identification of pathogens, and the use of qPCR has become routine in diagnostic laboratories. The reliability of qPCR-based assays relies on a combination of optimal sample selection, assay design and validation as well as appropriate data analysis and the “Minimal Information for the Publication of real-time PCR” (MIQE) guidelines aim to improve both the reliability of assay design as well as the transparency of reporting, essential conditions if qPCR is to remain the benchmark technology for molecular diagnosis.

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