Biochemical Specimens
Cytogenetics Specimens
Chromosomal Microarray Analysis
DNA Specimens
Mitochondrial Specimens
Instructions for submission of products of conception and autopsy specimens

Biochemical Specimens:

Plasma: Send 1ml plasma (use green top, sodium heparin, tube). Although not required for testing, it is recommended that the sample be taken after a four hour fast. After separation of the plasma, maintain refrigerated and transport on cold packs to the laboratory between 8:00 am and 5:00 pm on the day of collection. Samples can be stored in the freezer overnight for transport the following day. Samples sent to the laboratory via overnight carrier should be sent frozen (freezer packs or dry ice) in an insulated container.

Serum: Send 1ml serum. Although not required for testing, it is recommended that the sample be taken after a four hour fast. After separation of the serum, maintain refrigerated and transport on cold packs to the laboratory between 8:00 am and 5:00 pm on the day of collection. Samples can be stored in the freezer overnight for transport the following day. Samples sent to the laboratory via overnight carrier should be sent frozen (freezer packs or dry ice) in an insulated container.

Cerebrospinal Fluid: Send 1ml CSF. After collection of the CSF, maintain refrigerated and transport on cold packs to the laboratory between 8:00 am and 5:00 pm on the day of collection. Samples can be stored in the freezer overnight for transport the following day. Samples sent to the laboratory via overnight carrier should be sent frozen (freezer packs or dry ice) in an insulated container.

Urine: Send 5-10 mls of random urine. Although not required for testing, it is recommended that the sample be taken from the first morning void to ensure that the specimen is sufficiently concentrated. After collection of the urine, maintain refrigerated and transport on cold packs to the laboratory between 8:00 am and 5:00 pm on the day of collection. Samples can be stored in the freezer overnight for transport the following day. Samples sent to the laboratory via overnight carrier should be sent frozen (freezer packs or dry ice) in an insulated container. Exception: All urines for urine succinylacetone MUST be frozen immediately after collection and maintained frozen or on wet ice at all times due to temperature instability.

Red Blood Cell Enzyme Assays: Draw 3-5 ml in a yellow top (ACD) tube and ship on wet ice in an insulated container by overnight express. Sample must arrive at our location within 48 hrs.

White Blood Cell Enzyme Assays: Draw 7-10 ml in a yellow top (ACD) tube and ship on wet ice in an insulated container by overnight express. Sample must arrive at our location within 48 hrs.

Cystine in White Blood Cells : Send 7-10 ml of heparinized blood (green top tube) on wet ice in an insulated container by overnight carrier. Sample must arrive at our location within 48 hrs.

Cultured Cells: Two T-25 flasks of confluent cells (skin, amniotic or chorionic villi cells). Send at room temperature in insulated container by overnight carrier.

Liver or Muscle: Send >10 mg of tissue, rapidly frozen following collection. DO NOT add any liquid to the specimen. on dry ice in insulated container by overnight carrier

Tissue Requirements:

• Skin Biopsy:
  Biopsy specimens are best taken by punch biopsy to include full thickness of dermis. Biopsy samples should be at least 3mm X 5mm in area. Aseptically place biopsy in a screw-capped, sterile container with sterile culture medium (sterile saline can be used if no media is available) at least 2/3 full. Tightly cap the tube and wrap with parafilm. The tube should be wrapped with packing material to prevent accidental breakage during shipping. The tube should be placed inside a sealable plastic bag before being placed inside the shipping container.


• Skin Fibroblast Culture:
  Two 90% confluent T-25 culture flasks should be sent and back-up cultures maintained at the referring institution until results are received. Completely fill the two flasks to the base of the neck with media prior to shipping. Tightly cap the flasks and wrap with parafilm. The flasks should be wrapped with packing material to prevent accidental breakage during shipping. The flasks should be placed inside a sealable plastic bag before being placed inside the shipping container.

Cultured Amniocytes and Chorionic Villus for Prenatal Analysis:
Two 90% confluent cultures in T-25 flasks should be sent (with a control flask if possible) and back-up cultures maintained at the referring institution until results are obtained. Completely fill the two flasks with media prior to shipping. Tightly cap the flasks and wrap with parafilm. The flasks should be wrapped with packing material to prevent accidental breakage during shipping. The flasks should then be placed inside a sealable plastic bag before being placed inside a shipping container.

Note: All prenatal samples should be shipped at ambient temperature in an insulated container. Please label all samples with name and date of birth with indelible ink. Do not ship specimens on Thursday or Friday unless the laboratory is notified first.

Cytogenetics Specimens

Ship at room temperature.
Do not heat or freeze.

Fresh whole blood in Sodium Heparin (green-top) tubes:
Newborn (< 3 months): Minimum 1.0 -2.0 cc
Adult/Child: Minimum 3.0 -5.0 cc
Specimens must be sent the same day as collected and arrive in the laboratory within 48 hours of venipuncture.

Amniotic Fluid:
Direct: 20-30 cc of fluid collected in two sterile 15 ml conical centrifuge tubes. Discard the first 2 cc of fluid collected.
Culture: 2 T25 flask grown to 60-70% confluency.

Chorionic Villus: 15 mg in sterile tissue culture media.

Tissue: 5 mm of specimen (skin, placenta, cord) in sterile tissue culture media.

Chromosomal Microarray Analysis (CMA)

POSTNATAL:

Blood in both EDTA (purple top) and Sodium Heparin (Green top) tubes:
Adult: 10 cc per tube
Child: 4 cc per tube
Infant: 2 cc per tube

PRENATAL:

1. Amniotic Fluid ( two options available):
   • Option I: CMA and cytogenetic studies at BCM lab: 25-30 cc of direct fluid (gestational age >16 weeks) (Discard first 2 cc)
   • Option II: CMA only at BCM lab with cytogenetic studies elsewhere: minimally 15 cc of direct fluid (gestational age >16 weeks) (Discard first 2 cc)
     
2. CVS – Entire fetal sample must be submitted for both CMA and cytogenetic studies to BCM lab. Estimated 20-30 mg of chorionic villi (undissected) should be sent in sterile media.
   
3. Fetal Blood Sample - Please contact the lab to discuss before collecting specimen
   
   Parental Bloods: 5-7 CCin EDTA tube on each parent.
   Note: Blood samples from both parents are required. Fetal sample will not be processed unless maternal blood sample is received with prenatal sample and paternal sample is either received with fetal sample or shipped in 48 hours or BCM lab is notified of unavailability of parental sample.
   

Instructions for submission of products of conception and autopsy specimens:

Less than 12 weeks gestation: Placenta (chorionic villi) and any identifiable fetal parts.

Greater than 12 weeks gestation:

1. 2-3 mL unclotted cord blood or cardiac blood in both EDTA and sodium heparin tubes for Chromosomal Microarray Analysis; only in sodium heparin for chromosome analysis.
2. 10x10x10 mm placenta
   - Sample fetal side near the site of the cord insertion.
3. Skin*
   - Obtain from a central location (eg. buttock or upper thigh) rather than from a distal location (eg. foot) to enhance cell viability and likelihood of results.
4. Lung*
5. Kidney*

*Specimens should be 5 mm x 5 mm x 5 mm or as large as feasible given fetal/patient size.

All tissue samples should be placed in a separate sterile container with RPMI media. Containers with media can be obtained from the Lab. In the absence of media, place specimen in a sterile container with a small amount of sterile saline. Never place samples in formalin or other fixative.

If fetal demise occurred more than 48-72 hours prior to retrieval of tissue and delivery to the Lab, fetal tissue is unlikely to grow. In the case of prior fetal demise, placenta is most likely to grow.

Please contact the lab at 713-798-6555 prior to ordering any DNA analysis on POC or autopsy specimens.

DNA Specimens:

Blood Specimens: EDTA (purple) tubes; Adult: 14 cc, Child: 6 cc, Infant: 2-3 cc

Fetal Specimens: Studies will be performed two separate times on either direct and culture, or two separate cultures. This may necessitate sending more than one set of cultures, so backups should be maintained until a final report is recieved.

Special Requirements for Fetal Specimens:
We require that the referring center consult with us regarding prenatal cases prior to submitting any samples. For the most accurate fetal results, analysis of family members is often required. For autosomal recessive conditions, blood specimens should be submitted from both parents and the affected sibling for molecular analysis. For autosomal dominant or X-linked conditions, the high risk parent and the affected relative should be studied. Some disorders may require analysis of additional family members for precise interpretation of fetal results. Fetal specimens should be clearly labeled with mother's full name, type of fetal sample (CVS or amnio), and the fetal sex determined by cytogenetic data. All fetal studies will be performed two times, on different assays, on either a direct and then a cultured specimen, or on two separate cultures. A preliminary report will be sent if results are obtained on the first study and this will be followed by a final report once the repeat study is complete. Maternal blood is required for all prenatal tests to rule out contamination and sample switch. Clearly indicate if ordering cytogenetics or biochemical studies in addition to DNA

• Amniotic Fluid:
  Direct: 10 cc unspun (not applicable for certain tests)
  Send: 30 cc for DNA + cytogenetics analysis or cell culturing
  Not applicable for Fragile X syndrome, Prader-Willi or Angelman methylation anaylsis.
  
• Chorionic Villi:
  Direct: 10 mg shipped in sterile transport media
  Not applicable for Fragile X syndrome, Prader-Willi or Angelman methylation anaylsis
  
• Culture (Both Amniocytes and CVS):
  2 T25 flasks with 100% confluent monolayer cultures
  2 T75 confluent flasks needed for DMD/BMD and Factor VIII cases
  Contact the laboratory for specific prenatal sample requirements.

Mitochondrial Specimens:

Label all specimens with the patient’s full name and date of birth. Enclose a completed mitochondrial requisition form, clinical summary or clinical check list, and signed consent form.

Use FedEX or UPS. for overnight shipment (see below).

Blood: EDTA (purple) tubes; Adults: 14 ml whole blood; Child: 6 ml whole blood; Infants: 2-3 ml whole blood
Ship at room temperature, do not heat or freeze.

Cultured Cells: Four T-25 flasks or 2 T75 of confluent cultured calls. Flasks should be filled completely with medium before shipping at room temperature.

Muscle biopsies: 50 mg for skeletal muscle DNA analysis, 150 mg for  skeletal muscle ETC analysis, >200 mg if additional tests are requested. Skeletal muscle and other tissues such as liver and kidney should be snap-frozen at collection, stored at -70°C, and shipped on dry ice, overnight delivery.

Other specimens accepted (for PCR based analysis only): urine sediment (ship with cold pack), hair bulbs, buccal cells, paraffin embedded tissues.