Detection and identification of fungal DNA in the Order Mucorales
Mucormycosis is caused by molds from the Order Mucorales. These fungi are ubiquitous in nature and are increasingly becoming recognized as opportunistic pathogens in immunocompromised or immunosuppressed patients. Infections are most commonly caused by Rhizopus oryzae, Rhizopus microsporus, Mucor spp, Rhizomucor sp. and Abisidia corymbifera. Mucorales fungi pose difficult diagnostic and therapeutic challenges because (i) the spectrum of opportunistic mucormycoses is expanding (ii) their clinical manifestations can be fatal without rapid diagnosis and treatment and (iii) strains that fail to sporulate under normal laboratory conditions may be encountered, thereby making morphological identification difficult.
The Molecular Microbiology laboratory designed a specific primer mix that can amplify most commonly encountered Zygomycetes i.e. Rhizopus oryzae, Rhizopus microsporus, Mucor spp. Rhizomucor sp. and Lichtheimia corymbifera. These specific primers are used in a nested PCR following amplification of the ITS2 region.
Ribes et al. (2000) Zygomycetes in Human Disease in Clinical Microbiology Reviews, pp236-301
DNA extraction, nucleic acid purification, polymerase chain reaction (PCR), sequencing
Ribes et al. (2000) Zygomycetes in Human Disease in Clinical Microbiology Reviews, pp236-301
Acceptable specimens are listed below. Please see Molecular Microbiology Specimen Submission for complete specimen collection and handling instructions.
Shipping/Handling
Acceptable Specimens
*Mycobacterium avium complex DNA Detection [MAVDNA] can be ordered on sputum
**Fungal PCR reflex NGS [FUNDNA] and Fungal DNA Detection by PCR (without reflex to NGS) [NRFDNA] may have interference due to some lots of eSwabs which have been found to contain Saccharomyces cerevisiae DNA, resulting in false positive detection. Clinical correlation and/or retesting with a different collection method is advised. The detection of S. cerevisiae from eSwab specimens can interfere with our ability to rule out other fungal DNA.
Unacceptable Specimens
Optimal Quantity:
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.
Fresh tissue is the optimal specimen of choice, as it reduces the chance of introducing exogenous DNA templates or microorganisms during embedding/fixation. Formalin fixation dramatically reduces the sensitivity of the assays due to reduced template yield and quality.
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Please see Molecular Microbiology Specimen Submission for complete specimen collection and handling instructions.
UWMC/HMC: Store and send fresh tissue/fluid specimens refrigerated, if specimen storage and transport will exceed 8 hours, freeze at -20°C. Freeze all fresh tissue/fluid specimens at -20°C upon arrival in UW Molecular Microbiology.
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 |
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