Aspergillus fumigatus DNA detection by PCR
General Information
Lab Name
Aspergillus fumigatus DNA detection by PCR
Lab Code
ASPDNA
Epic Name
Aspergillus fumigatus DNA detection by PCR
Description
Detection of Aspergillus fumigatus DNA.
Aspergillus fumigatus is considered the most prevalent airborne fungal pathogen that causes severe and often fatal invasive aspergillosis (IA) among immunocompromised individuals such as patients with leukemia or those who have undergone organ transplantation.
A rapid test with high sensitivity and specificity for detecting A. fumigatus DNA in clinical specimens is required for successful antifungal therapy.
Internal Transcribed Spacer sequence (ITS2) between the 5S and 28S genes exhibit species-specific polymorphisms suitable for distinguishing A. fumigatus from other fungi including genetically closely related species such as Neosartorya ficheri. A real time PCR test that can detect the presence of A. fumigatus DNA in BAL or biopsied tissue was developed on a Lightcycler (Roche) platform. A nested PCR protocol with A. fumigatus specific FRET probes is employed to achieve high sensitivity and specificity.
For more information on this test, to download a requisition form, and for a list of acceptable and unacceptable specimens, please refer to: Molecular Diagnosis Microbiology Section Website
Synonyms
Aspergillosis PCR, Aspergillus fumigatus detection, Aspergillus fumigatus identification, Aspergillus fumigatus PCR, Aspergillus identification, Aspergillus PCR, molecular Aspergillus, universal PCR
Components
Interpretation
Method
DNA extraction, nucleic acid purification, polymerase chain reaction (PCR), sequencing
Reference Range
See individual components
Guidelines
Ordering & Collection
Specimen Type
Collection
Acceptable specimens are listed below. Please see this page for complete specimen collection and handling instructions.
Specimens should be collected into a DNA free container labeled with at least two identifiers.
Acceptable Specimens
- Fresh frozen tissue
- Fresh frozen fluid: any body fluid is acceptable if it is not listed under Unacceptable Specimens. See common examples below.
- Formalin Fixed Paraffin-embedded tissues (FFPE, PET): blocks, scrolls, and unstained slides are acceptable
- eSwabs and UTM (universal transport media)
- Sodium polyanethol sulfonate (SPS, Wampole Isolator Tubes) acceptable with disclaimer
Common acceptable body fluid examples: cerebrospinal fluid, pleural fluid, pericardial fluid, urine, bronchial lavage, joint fluid, bone marrow, vitreous fluid, etc.
Additional Acceptable Specimens
- Sputum
Unacceptable Specimens
- Blood, serum, plasma, stool
- No citrated or heparinized solutions
- Tissues floating in formalin
- Swab/fluid collected in tube containing agar
Handling Instructions
Fresh frozen tissue/fluid specimens should be submitted and maintained on dry ice.
Formalin Fixed Paraffin-embedded tissues (FFPE, PET) can be sent ambient or with ice packs during warmer summer months to prevent melting.
Optimal Quantity:
- Fresh Tissue: 0.3-1.0 cm^3
- Fluid: 0.2-1 mL
- Formalin Fixed Paraffin-embedded Tissue (FFPE/PET): blocks are preferred and will be sent back to client upon completion of testing
- Scrolls/unstained slides: cross-sectional area >1cm^2 send 10 sections of 10µm thickness, if <1cm^2 send 20 sections if available
Please note: We do not need a separate specimen aliquot for each test ordered. Only a single specimen aliquot or block of optimal quantity is necessary for performing multiple tests. If multiple aliquots or blocks of optimal quantity are sent, up to 2 will be pooled.
Quantity
requested: See Optimal Quantity above
minimum: Specimens below optimal quantity are acceptable for testing, however, diagnostic yield is generally proportional to specimen size.
Processing
UWMC/HMC – store and send specimen refrigerated and freeze specimens at -20°C upon arrival in UW-ML Microbiology. If specimen storage and transport will exceed 8 hours, freeze at -20°C.
Freeze specimens at -20°C upon arrival
Outside Laboratories: Freeze specimens in sterile containers at -20°C. Transport all specimens on dry ice.
Performance
LIS Dept Code
Micro Molecular Diag (MMD)
Performing Location(s)
| UW-MT |
Microbiology, Molecular Diagnostics
206-520-4600 ---------------------------------------- Shipping Address Attn: Molecular Microbiology Performing Lab Address Clinical Microbiology Lab, NW177 |
Contact Information Please e-mail us with any questions or comments you may have. Your inquiry will be answered as soon as possible. email: molmicdx@uw.edu The Molecular Microbiology lab is open from Monday-Friday, 7am-4pm PDT. Billing inquiries and requests for faxed reports can be made to our Client Services Department at (206) 520-4600 or (800) 713-5198. For results or other inquiries, we can be reached by phone at the following numbers:
For assistance during weekends, holidays and after hours, please contact Lab Medicine Resident at (206) 598-6190 |
|---|
Frequency
Fresh frozen tissues/fluids result in 2-3 business days after receipt of specimen. Formalin Fixed Paraffin-embedded tissues result in 3-4 business days after receipt of specimen.
Available STAT?
No