Background:  Fungal diseases are mostly opportunistic infections with a great variety of common and uncommon pathogens. Up to 150 fungal species have been shown as potential human pathogens involving all body sites. Invasive mycoses are increasingly recognized as a primary cause of morbidity and mortality especially in immunocompromised patients. These infections are mainly caused by yeasts (such as Candida spp., Cryptococcus neoformans, Saccharomyces cerevisiae, Trichosporon spp.), moulds (such as Aspergillus spp., Scedosporium spp., Fusarium spp., Natrassia mangiferae, Curvularia spp., Schizophyllum commune, Paecilomyces variotii, Bipolaris spp., Cladophialophora bantiana) and zygomycetes (such as Rhizopus spp., Absidia spp., Rhizomucor spp., Mucor spp., Cunninghamella bertholletiae, Saksenaea vasiformis, Apophysomyces elegans, Basidiobolus ranarum, Conidiobolus spp.). The mortality rate of invasive fungal infections is 40-100%. A rapid diagnosis improves the outcome. Non-invasive fungal infections (such as infections of the urogenitary tract, of the eye, of the skin, etc.) are mainly caused by Candida, Aspergillus, Acremonium, Fusarium and dermatophytes (such as Trichophyton spp., Microsporum spp.).

Description:  MycoReal® Kit Fungi is a real-time PCR for the qualitative detection of the DNA of fungi (ITS2-region). For phylogenetic identification of the pathogen, the PCR amplicon must be sequenced and analysed by BLAST. MycoReal® Kit Fungi does not displace culture but offers an improvement in the detection of fungi in cases where the presence of fungi is suspected but culture remains negative, or the fungus is difficult to cultivate. This kit is suitable for specimens taken from normally sterile sites. Proper specimens are DNA extracts isolated from samples of bronchoalveolar lavages (BAL), aspirates, cerebrospinal fluid, tissue, paraffin embedded tissue, fungal colonies as well as of environmental samples. The kit is suitable to a limited extent for the detection of fungi in blood samples. In case of a mixed fungal infection, the pathogen cannot be phylogenetically identified.

MycoReal® Kit Fungi
Order Number Reactions Channel Pathogen Channel IPC Target
DHUF00453 50 FAM Cy5 ITS2
For research use only
Shipping Temperature: -20 °C to +4 °C
Product Features:
  • Amplification and detection: ITS2 region of fungi
  • Real-time PCR with rapid hot-start Taq DNA polymerase
  • ROXdye as passive reference
  • Internal Positive Control System to exclude false-negative results
  • Optimized to handle PCR inhibitors
  • PCR platforms: runs on all established standard real-time PCR platforms
  • Harmonized thermal profiles to run RNA and DNA samples simultaneously

PCR platforms: MycoReal® Kit Fungi has been validated with the ABI® 7500 Real-time PCR System (Thermo Fisher Scientific, fast cycle parameters are not supported) and with the cobas z 480 Analyzer (Roche). It is also suitable for other real-time PCR instruments that can measure and differentiate fluorescence in the FAM and Cy5 channel (e.g. QuantStudio5, QuantStudio7 real-time PCR system (Thermo Fisher Scientific), qTOWER3G (Analytik Jena), Mic instrument (bio molecular systems), LightCycler® 480 II (Roche Diagnostics)).

Frequently Asked Questions:

The kit enables universal detection of fungal DNA by targeting the ITS2 region. It is intended for broad-range screening of fungi in clinical and environmental samples.
Validated sample types include bronchoalveolar lavage, aspirates, cerebrospinal fluid, fresh or paraffin-embedded tissue, fungal colonies, and to a limited extent EDTA blood.
The PCR amplicon of approximately 370 bp can be sequenced and analyzed by BLAST against NCBI databases to enable phylogenetic identification of the fungal species.
An internal positive control (IPC) is added during DNA extraction and detected in the Cy5 channel. It monitors extraction efficiency and identifies PCR inhibition.
Broad-range PCR enables detection of a wide variety of fungi, including difficult-to-culture organisms. However, due to potential contamination with environmental fungi, results require sequencing confirmation and careful clinical interpretation.
BactoReal®, MycoReal®, ParoReal® and ViroReal® Kits run with the same thermal cycling conditions.