Indications: There is a large variety of clinical manifestations associated with defects of electron transport ("respiratory") chain, as the components are encoded by many nuclear and mitochondrial DNA genes. Blood and/or CSF lactate may be increased with a high lactate/pyruvate ratio, and urine organic acid or acylcarnitine analyses may show evidence of impaired fatty acid and amino acid oxidation. Muscle biopsies may show proliferation and/or abnormally shaped mitochondria. Available assays include, initially, rotenone-sensitive NADH-cytochrome c reductase (complexes I + III), antimcycin-sensitive succinate cytochrome c reductase (complexes II +III), decylubiquinol cytochrome c reductase (complex III), NADH ferricyanide reductase (part of complex I), succinate dehydrogenase (part of complex II), and cytochrome c oxidase (complex IV). If these results are abnormal, follow-up testing includes complex II and, in isolated mitochondria, complex I. These defects can, in some cases, be detected by assays in cultured skin fibroblasts or liver(complexes II-IV), but assays of skeletal muscle or heart, if available, are more valuable (complexes I-IV). Mitochondrial DNA analysis should be considered as part of the evaluation of these defects.
Method Description: Spectrophotometric enzyme assays: cytochrome c reduction with NADH + rotenone (I + III), succinate + antimycin (II +III), decylubiquinol + antimycin (III), oxidation of reduced cytochrome c (IV).
| Specimen |
Specimen Requirements |
Shipping Requirements |
Skeletal Muscle or
Heart |
After a biopsy/autopsy, the muscle should be immediately quick-frozen in liquid nitrogen.
Minimum Amt Required: 100 mg |
Ship on dry ice. If biopsy is performed on a Friday, keep sample at -70°C or colder over the weekend and ship on Monday. |
| Liver |
After a biopsy/autopsy, the muscle should be immediately quick-frozen in liquid nitrogen.
Minimum Amt Required: 100 mg |
Ship on dry ice. If biopsy is performed on a Friday, keep sample at -70°C or colder over the weekend and ship on Monday. |
| Cultured Skin Fibroblasts |
REQUIRES ADVANCE SCHEDULING
Two confluent T25 flasks filled to the neck with media in a non-vented flask and tape/parafilm shut. If several assays are ordered (more than 3), the shipment of an additional T25 flask is recommended.
Also the cell line MUST be MYCOPLASMA tested. If your institution is unable to test for mycoplasma, please tell us when you schedule a shipment or send them to Bionique to get tested.
Amt Required: 2T25 flasks |
Schedule a shipment time and ship the 2T25 flasks at room temperature with enough packing material so flasks do not leak.
Please notify us if you are unable to ship the skin fibroblasts on your scheduled shipment time. |
| Skin Biopsy |
Biopsy must be obtained under sterile conditions. Be careful to remove all betadine and alcohol and rinse with saline prior to obtaining the sample. Use a 3 or 4mm punch biopsy kit or take from the site of incision for a muscle biopsy.
Place biopsy in a container of sterile Hanks Balance Salt Solution (HBSS), sterile Ringer's Lactate, or other physiological solution.
Minimum Amt Required: a bx punch or a piece |
Call us to let us know you'll be sending the skin biopsy and ship at room temperature with enough packing material so container does not leak.
|
Samples Accepted: Monday - Friday, 8:30 AM - 5:00 PM
Turn-Around-Time:
2-3 weeks for skeletal muscle, heart, liver
3-4 weeks for skin fibroblasts cultured from T25
7-10 weeks for culturing skin fibroblasts from a skin biopsy
(skin biopsies grow at different rates and sometimes can
take 3 months or longer)
CPT Codes for assay of fibroblasts or tissues: 84999, 82657, 84157, 80502
CPT Codes for Cell Culture: 88233, 88240
CPT Code for Mycoplasma testing if your institution is unable to do this: 86738
Specimens can be sent via Federal Express, DHL, or UPS using Overnight Priority/Next Morning service.
Special Instructions for skin fibroblasts: Ship T25 flasks with Mycoplasma results. Also save back-up flasks of skin fibroblast culture(s) at your institution until you have received the test results in the event of breakage of flasks or contamination during transport.
Future Testing on skin fibroblasts: We freeze down plugs on the cell line(s) you send to us, so if additional testing needs to be ordered in the future, we will have plugs here at CIDEM to initiate.
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