Duchenne Muscular Dystrophy (DMD), Gene Deletion/Duplication and Sequencing (Sendout)

Test Description: This test provides full coverage of all coding exons of the DMD gene, plus ~10 bases of flanking noncoding DNA. Full coverage is defined as >20X NGS reads or Sanger sequencing.

• PreventionGenetics Test Description: Dystrophinopathy via the DMD Gene

Indications for Test:

• Males with a clinical diagnosis of DMD/BMD.
• Males with symptoms of DMD/BMD (a waddling gait and difficulty climbing stairs, Gower sign, elevated CPK).
• Male/Females with a negative DMD/BMD test by multiplex PCR, MLPA, or Southern blot.
• Males/Females with previous DMD/BMD test result with unclear del/dup size and boundary.
• Females at risk of being a carrier (Previous child or a family history of DMD/BMD).

Ordering Requirements:

• Orders must be accompanied by a completed PreventionGenetics Standard Requisition or a copy of an order completed in the Prevention portal.

Whole Blood:

• 3-5 mL whole blood in LAVENDER TOP (EDTA) tube or YELLOW TOP (ACD)
• Pediatric: 1 mL whole blood

Cell Culture: 2 T25 flasks of confluent cells.

Direct Amniotic Fluid or Chorionic Villus Sampling (CVS): 10-20 mL of direct amniotic fluid or 5-10 mg cleaned CVS tissue in sterile media or saline. A back-up culture is required for ongoing pregnancies.

DNA:

• 5 µg -10 µg of purified genomic DNA at a concentration of at least 100 ng/uL in a screw cap tube.
• Indicate composition of the solute and DNA concentration on the label, along with the patient's name, date of birth, and/or ID number.
• Only genomic DNA will be accepted. Prevention does not accept products of whole genome amplification reactions or other amplification reactions.
• DNA must be extracted from a CLIA-certified laboratory or a laboratory meeting equivalent requirements as determined by CAP and/or CMS.

Fresh Tissue/POC: 15-20 mg or 2 mm x 2 mm x 2mm tissue.

• Testing on direct tissue: Flash freeze. Tissue should be sent frozen (preferably on dry ice).
• Tissue for cell culture: Send in sterile media or saline (not frozen). Contact Prevention Genetics at (715) 387-0484 for additional detail.

Saliva/Buccal: Oragene™ or GeneFix™ saliva collection kit or OCD-100 buccal swab collected according to manufacturer instructions.

• Note: UW-MT DLMP does not stock collection kits. Providers must contact PreventionGenetics directly to obtain a kit.

Outside Laboratories:

• Store and transport whole blood at ambient temperature.
• Submit a completed a PreventionGenetics requisition along with the DLMP requisition*.
  • *Use client-specific DLMP requisition when available. To obtain a client-specific DLMP requisition, contact Client Support Services via email at: commserv@uw.edu.

Stability (Whole Blood): Ambient (preferred): 8 days; Refrigerated: 8 days; Frozen: 1 month.

Store and transport sample(s) at ambient temperature unless otherwise indicated.

• Other sample types:
  • DNA, Cell Cultures, Saliva, Buccal Swabs, Direct Amniotic Fluid/CV/POC: Ambient temperature.
  • Direct Tissue: Freeze at -20°C.

Login: RPREVS-;AMBIENT

• RPRCOD: ;Prevention Genetics Test 1773
• RPRTST: ;Dystrophinopathy via the DMD Gene
• RPRVSP: [Enter the specimen type using the codes below.]

Specimen Type Codes:

• WB (Whole Blood)
• DNAEXT (Extracted DNA)
• BNM (Bone Marrow)
• BUCSWB (Buccal Swab)
• CV (Chorionic Villi)
• AMN (Amniotic Fluid)
• CULAMN (Cultured Amniocytes)

Sendouts:

• Order Prevention Genetics Test: 1773.
• Ship the sample with a completed requisition Monday through Friday via FedEx Priority Overnight. Shipments sent on Friday must be marked for Saturday Delivery.

Stability (Whole Blood): Ambient (preferred): 8 days; Refrigerated: 8 days; Frozen: 1 month.