Neuromuscular Center /Biochemistry Lab

Institute for Exercise and Environmental Medicine

Presbyterian Hospital of Dallas

7232 Greenville Ave, Dallas, TX 75231

Tel : (214)345-4611                                                                                                                                                                                                                 CLIA ID No 45D0887428

FAX : (214)345-7105                                                                                                                                                                                                                         CAP No 20697-09

SPECIMEN INFORMATION SHEET

PURPOSE for requesting a Mitochondrial Myopathy Enzyme Panel Analysis :

Mitochondrial defects may result from mutations in either mitochondrial or nuclear DNA and often result in deficiency of enzymes of the respiratory chain. The resulting biochemical abnormalities may consist of a single enzyme deficiency when there is a nuclear DNA defect or, a deficiency involving single or multiple enzymes when the genetic defect involves mitochondrial DNA. The Mitochondrial Myopathy Enzyme Panel is designed to assess individual enzymatic activities of Complex I, II, III and IV and to assess the level of activity of the marker mitochondrial enzyme, citrate synthase. Selective or multiple deficiencies of these enzymes may permit the diagnosis of a mitochondrial myopathy and provide clues to molecular pathogenesis.

The symptoms of such metabolic diseases of skeletal muscle include: muscle weakness, particularly weakness of the eye lids (ptosis) and of the extraocular muscles; exercise intolerance/exertional muscle fatigue, often associated with prominent exertional shortness of breath; exertional muscle pain may be present but exercise induced cramps are uncommon. In some cases, mitochondrial myopathies are associated with episodes of rhabdomyolysis and myoglobinuria.

Conditions For REJECTION: To ensure the tests will be conducted with accuracy and the highest quality, muscle biopsies will be considered unacceptable under the following circumstances:

The muscle arrives thawed.
The shipping box is damaged and the sample has been destroyed.
The muscle biopsy is under the minimum amount required (75 mg).
The sample vial is unlabeled.

PLEASE NOTE: An incomplete requisition form will result in delay of testing and result reporting, and, may be a motive for rejection. Oral request for testing must be followed by a written request within 30 days and before the report can be released.

Muscle Specimen Freezing Procedure:

A punch biopsy or needle biopsy will provide between 75 and 150 mg of muscle. In case of open biopsy, excise a specimen approximately 0.5 cm x 0.5 cm x 0.5 cm with minimal trauma, by dissecting along the long axis of the muscle.
Optimally, flash freeze the piece(s) of muscle in liquid nitrogen on site immediately after excision. If you need to transport the biopsy to another site for weighing or freezing, you can use a sterile plastic Petri dish (or other sterile disposable plastic container) containing some gauze slightly moistened (not dripping) with sterile saline (0.9% NaCl). You should keep the Petri dish on ice.
Manipulate the muscle biopsy with care using sterile forceps.
Flash freeze by plunging the specimen into liquid nitrogen for 10 to 20 sec. Be aware that the specimen will fall to the bottom of the container when it is properly frozen. MAKE SURE to use a container that allows you to retrieve the specimen. The well-frozen specimen has a white chalky color.

Note: If you do not have access to liquid nitrogen, you still need to freeze the muscle specimen AS FAST AS POSSIBLE. One possibility is to use dry ice in ethanol 95%. DO NOT immerge specimen in ethanol; place specimen in vial (test empty vial to make sure it stands ultralow temperature), and immerge the bottom of uncapped vial in the cold mixture. DO NOT LET THE SPECIMEN IN CONTACT WITH THE ETHANOL MIX. Cap and keep frozen. Another way is to place small pieces of muscle on a piece of dry ice, then scrape them off, transfer in COLD vial, and keep frozen immediately (-70° C). TO AVOID FREEZE/THAW, never touch a frozen specimen with room temperature "utensils" (forceps, vial, fingers, etc…)!

Immediately after freezing, plunge one LABELED cryovial into the liquid nitrogen (write at least name of patient and date on the vial using an indelible marker Sharpieä type) and transfer the specimen into the vial. If you use forceps be sure to plunge the tip in the liquid nitrogen BEFORE touching the frozen specimen. Failing to do so will result in local thawing which may compromise the results.
Carefully screw cap on cryovial. The vial can either stay in liquid nitrogen (if shipment on dry ice follows) or it can be stored below -70° C in a deep freezer or on dry ice until shipment.
To avoid compromising the detection of muscle enzymes, it is extremely important to keep the time between collection and freezing as short as possible. To our knowledge, NADH Ubiquinone Reductase (NUR) which is the most sensitive enzyme in the mitochondrial myopathy panel, will remain stable for 1 hour. Delays in freezing may compromise the detection of Complex I (NUR) deficiency.

Thawing and refreezing will definitely compromise the detection of ALL enzymes.

If the sample is not sent out the day it is collected and frozen, it is critical to store it frozen BELOW -70° C. Keeping the vial in a regular -20° C may compromise the analysis of enzymatic activities.

Turnover Time :

The biochemistry lab will provide an interpretive report. Normal turnover time is 2 to 3 weeks. If emergency analysis is required, please call PRIOR to the biopsy.

SHIPPING and HANDLING INFORMATION :
A minimum of 75-100 mg of muscle (size equivalent to a chick pea) is required for the mitochondrial myopathy enzyme panel. The inclusion of significant amounts of fat or connective tissue in the biopsy will decrease the accuracy of the test.
Immediate freezing is absolutely necessary. The use of liquid nitrogen is highly recommended. Please fill out the time of collection and the time of freezing on the request form. Failing to do so will compromise the results especially for Citrate Synthase and Complex I (NUR).
Ship with 7 pounds of dry ice or store at -70° C or lower until shipment. Do not treat with fixatives or saline solution. Biopsies once frozen cannot be thawed under any circumstances because of potential degradation of muscle enzymes.
Make sure to send sample using OVERNIGHT PRIORITY/NEXT MORNING service offered by your courier (FEDEX, Airborne Express, or UPS are possible choices). Make sure to fulfil all requirements for dry ice shipment. The styro-foam boxes MUST be IN a cardboard box with a special label. You may need to contact the chosen company for more information about dry ice overnight shipment if necessary.
Include the Request Form in a sealed plastic bag inside the shipping box.
Please send sample ONLY from Monday through Thursday, not before a holiday and make sure to notify the lab that there is a shipment expected.
We do not provide any shipping material and will not pay any shipping charges.

For any "special" requirement or more information, please don’t hesitate to call PRIOR to the procedure and ask for Dr. Nadine Romain, Ph.D. for technical information.

Print out and complete form below

 

Neuromuscular Center/ Biochemistry Laboratory                                              Lab USE ONLY

Institute for Exercise and Environmental Medicine                                                   LAB. ACC. # ___________________

Presbyterian Hospital of Dallas                                                                             Date received ___________________

7232 Greenville Ave, Dallas, TX 75231                                                                  Time received ___________________

Ronald G. Haller, MD, Director

Tel : (214)345-4611                         Mitochondrial Myopathy Enzyme Panel                     CLIA ID No: 45D0887428

FAX : (214)345-7105                                     REQUEST FORM                                                       CAP No: 20697-09

(PLEASE PRINT)

PATIENT’S NAME: (Last)

(First)

DATE OF BIRTH:

PARENTS’ NAME (If child):

PATIENT SS#:

HOSPITAL/CLINIC/LAB:

SEX:

HOSPITAL/CLINIC/LAB TELEPHONE:

PATIENT MED REC.:

ORDERING PHYSICIAN(S):

OFFICE ADDRESS:

TEL:

FAX:

SEND BILL TO:

COMPANY/HOSPITAL NAME:

ADDRESS:

CITY:

STATE:

ZIP:

BILLING CONTACT PERSON:

TEL:

DIAGNOSIS & ICD-9 CODE:

FAX:

COMMENTS:

SPECIMEN: DATE COLLECTED:

  

SPECIMEN FROZEN IN LIQUID NITROGEN?:

˜ YES ˜ NO

TIME COLLECTED:

 

TIME FROZEN:

  

MUSCLE BIOPSIED:

 

CLINICAL HISTORY (e.g. ophthalmoplegia, fatigability, myoglobinuria,...):

RELEVANT LABORATORY FINDINGS (e.g. Resting Lactate, muscle histology):

 

ANY FAMILY HISTORY OF MUSCLE DISORDER?

REQUESTED ANALYSIS (PLEASE, CHECK & SIGN)

 

˜ MITOCHONDRIAL MYOPATHY ENZYME PANEL (CPT CODES: 80050, 88319, 84311) including:

- CITRATE SYNTHASE (CS)

- UBIQUINONE CYTOCHROME C REDUCTASE (Complex III)

- NADH UBIQUINONE REDUCTASE (Complex I)

- CYTOCHROME OXIDASE (Complex IV)

- SUCCINATE DEHYDROGENASE (Complex II)

FEE: $1,500

REQUESTED BY: NAME (PLEASE PRINT)

 

 

                                                      SIGNATURE: ____________________________________ DATE: _____/____/_____

 
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7232 Greenville Avenue

 Dallas, Texas, 75231   (214)-344-4611