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Background: PanReal Kit Fungi & Bacteria is a multiplex-PCR real-time PCR test for universal detection of fungi and bacteria in samples purified from bronchoalveolar lavages (BAL), aspirates, cerebrospinal fluid, tissue, paraffin embedded tissue and fungal/bacterial colonies. Due to contamination, PanReal Kit Fungi & Bacteria is less sensitive compared to species-specific assays. The kit is suitable only to a limited extent for the detection of fungi in blood samples. This test is not suitable for the detection of bacteria in blood samples. For phylogenetic identification of the pathogen, the PCR product has to be sequenced and analysed by BLAST. In FAM channel the ITS2-region of fungi is detected (amplicon of about 370 bp). In VIC channel the 16S rDNA gene of bacteria is detected (amplicon of about 550 bp). The internal DNA positive control (DNA IPC) is detected in Cy5 channel (667 nm) and is used as DNA extraction as well as real-time PCR inhibition control. The target for the DNA IPC is extracted with the sample.
Description: 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 and moulds. 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. Concerning bacterial infections, broad‐range PCR targeting the 16S rDNA gene is useful when antimicrobial therapy has already been initiated or when culture remains negative due to infections caused by bacteria with unusual growth requirements.
Order Number | Reactions | Channel Pathogen | Channel IPC | Target |
---|---|---|---|---|
DHUFB0153 | 50 | FAM, VIC | Cy5 | ITS region/16S rDNA |
- Amplification and detection: ITS2 region of fungi and 16S rDNA of bacteria
- Real-time PCR with rapid hot-start Taq DNA polymerase
- ROX™ dye 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: PanReal KitFungi & Bacteria is developed and validated for the Applied Biosystems® 7500 System (Thermo Fisher Scientific), LightCycler® 480 (Roche) and Mx3005P® QPCR System (Agilent), but is also suitable for other real-time PCR instruments.