M. tuberculosis complex DNA Detection
General Information
Lab Name
M. tuberculosis complex DNA Detection
Lab Code
TBCDNA
Epic Name
M. tuberculosis complex DNA Detection
Description
Detection of Mycobacterium tuberculosis complex DNA.
Although an acid fast smear (AF smear) can provide an important clue to the nature of the causative agent for a patient's disease, a positive result does not necessarily indicate the presence of Mycobcaterium tuberculosis complex (MTBC). The presence of Acid Fast Bacillis (AFBs) in clinical specimens can be simply due to the growth of common water contaminants such as M. chelonae and M. fortuitum. On the other hand, a negative smear result also does not rule out the presence of M. tuberculosis complex at low pathogen load. Recent developments in PCR based amplification and detection of organism specific DNA offers an alternative that can be more specific and sensitive than a smear test.
For an accurate detection of the presence of MTBC in clinical specimens, the UWMC Molecular Diagnosis Section utilizes a multiplex PCR protocol that targets insertion elements IS6110 and IS1081, which are considered marker sequences for the identification of Mycobacterium tuberculosis complex members. Utilization of gene specific multiplex primers on a real-time PCR platform facilitates rapid and highly sensitive detection of M. tuberculosis complex-specific DNA in clinical specimens.
In many specimens acid fast bacilli can be seen microscopy of tissue sections but are very difficult to grow due to their fastidious nature, or are not viable as a result of antimicrobial therapy. Some specimens may never reveal the presence of a pathogen because of low abundance and/or lack of viability. The use of PCR to detect this DNA extracted directly from clinical specimens facilitates the identification of these pathogens.
Synonyms
AFB PCR, broad range AFB PCR, broad range Mycobacteria PCR, IS1081, IS6110, M. tuberculosis complex, Molecular AFB, Molecular Mycobacteria, Molecular Mycobacterium, MTB, MTB PCR, MTBC, multiplex PCR, Mycobacteria PCR, Mycobacterial identification, Mycobacterium identification, Mycobacterium tuberculosis complex identification, Mycobacterium tuberculosis complex PCR, TB PCR, TBC, Tuberculosis PCR, universal AFB PCR, universal Mycobacterium 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 Molecular Microbiology Specimen Submission for complete specimen collection and handling instructions.
Shipping/Handling
- Fresh frozen tissue/fluid specimens should be collected into a DNA free container labeled with at least two identifiers and 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.
Acceptable Specimens
- Fresh frozen tissue
- Fresh frozen fluid: any body fluid is acceptable if it is not listed under Unacceptable Specimens.
- Common examples include: cerebrospinal fluid, pleural fluid, pericardial fluid, urine, bronchial lavage, joint fluid, bone marrow, vitreous fluid, etc.
- Formalin Fixed Paraffin-embedded tissues (FFPE, PET): blocks, scrolls, and unstained slides
- Sputum: acceptable - except for Bacterial PCR reflex NGS [BCTDNA]/Bacterial DNA Detection by PCR (without reflex to NGS) [NRBDNA], Fungal PCR reflex NGS [FUNDNA]/Fungal DNA Detection by PCR (without reflex to NGS) [NRFDNA], and Nontuberculous Mycobacteria DNA Detection [NTMDNA]* assays
- eSwabs**, UTM (universal transport media), body fluid/bone marrow in EDTA (not including blood)
- Sodium polyanethol sulfonate (SPS, Wampole Isolator Tubes) acceptable with disclaimer
*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
- Blood, serum, plasma, stool/rectal swabs
- No citrated or heparinized solutions
- Tissues floating in formalin
- Swab/fluid collected in tube containing agar
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.
When possible, a mycobacterial culture is recommended regardless of the molecular test result to aid in clinical diagnosis.
Forms & Requisitions
Molecular Microbiology Order Form
Handling Instructions
Please see Molecular Microbiology Specimen Submission for complete specimen collection and handling instructions.
Quantity
requested: See "Collection" for Optimal Quantity
minimum: Specimens below optimal quantity are acceptable for testing, however, diagnostic yield is generally proportional to specimen size.
Processing
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.
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