The standard initial screening for a peroxisomal disorder

The standard initial screening for a peroxisomal disorder involves:

  1. Investigation of the presence/absence and morphology of peroxisomes by immunofluorescence;
  2. Investigation of very long-chain fatty acid metabolism;
  3. Depending on the indication additional tests can be performed such as single enzyme activity measurements (e.g. Dihydroxyacetonephosphate-acyltransferase, Acyl-CoA oxidase I, D-bifunctional protein, Sterol carrier protein X), immunoblot analysis or complementation analysis.

Price

€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Material:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.

Test request form Enzyme diagnostics

Acyl-CoA oxidase 1 (ACOX1)

Disease

Acyl-CoA oxidase 1 deficiency.

Description

Acyl-CoA oxidase 1 (ACOX1) is the first enzyme of the peroxisomal beta-oxidation pathway for straight-chain fatty acids.

OMIM

264470

Method

UPLC-UV

Price

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics:Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
C26:0-Lyso-PC

Disease/indication

X linked adrenoleukodystrophy (X ALD), Zellweger spectrum disorders, ACOX1 deficiency, DBP deficiency and ACBD5 deficiency.

Description

C26lysoPC is a marker for very long chain fatty acid accumulation in the phospholipid fraction and is increased in X linked adrenoleukodystrophy, Zellweger spectrum disorders, ACOX1 deficiency, DBP deficiency and ACBD5 deficiency.

Method

HPLC-MS/MS

TAT

4 Weeks. If the test is performed in cultured fibroblasts the TAT is 3 months

Price

€ 216
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

For requests in blood please use Test request form Metabolite diagnostics and for requests in skin fibroblasts Test request form Enzyme diagnostics.

Test request form Metabolite diagnostics

Test request form Enzyme diagnostics

Materials:

Blood spots

  • Amount: 2 dried bloodspot, 40 μl of blood on a guthrie card.
  • Conditions: Room temperature.
  • Specifics: Either capillary or venous blood spots can be collected. When preparing venous blood spots from EDTA-anticoagulated blood, please spot 40 μl blood per spot on a Guthrie card. The Guthrie card should be air-dried at room temperature for 4 hours.

Plasma

  • Amount: 500 μl.
  • Conditions: Dry ice.
  • Specifics: EDTA plasma (preferred material), heparin plasma also suitable.

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T 25 flask, fill the flask
    completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM)
    supplemented with 10% serum and 1% penicilline streptomycine. Cap the flasks tightly and wrap
    the cap with parafilm. Label the flask clearly with identifying information (name of patient and
    date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the
    Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to
    arrive in our laboratory within 48 hours. Please complete the test request form and send it along
    with the sample.If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you
    can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM
    or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin
    biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form
    and send it along with the sample.
Complementation testing

Disease

Zellweger spectrum disorders.

Description

Complementation testing is used to determine the defective PEX gene in fibroblasts from Zellweger spectrum disorder patients. Zellweger Spectrum disorders include Zellweger Syndrome, Neonatal AdrenoLeukoDystrophy (NALD) and Infantile Refsum Disease (IRD) and can be caused by mutations in any of at least 13 different PEX genes resulting in a defect in peroxisome biogenesis. Please note that before complementation testing can be performed, peroxisome deficiency has to be confirmed by catalase immunofluorescence microscopy analyses. After identification of the defective PEX gene by complementation analysis, molecular analysis of the candidate PEX gene is performed by our DNA diagnostic section.

OMIM

214100 202370

Method

Microscopy

Price

€ 1055

Test request form DNA diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
D-Bifunctional protein (DBP/MFP2)

Disease

DBP deficiency.

Description

D-Bifunctional protein (DBP/MFP2) catalyzes the second and third step of the peroxisomal beta-oxidation of straight-chain fatty acids and branched-chain fatty acids, including pristanic acid and C27-bile acid intermediates.

OMIM

261515

Method

UPLC-UV

Price

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.

Whole EDTA blood

  • Test material: Peripheral blood mononuclear cells isolated from whole blood.
  • Conditions: Room temperature.
  • Specifics: At least 4.5 ml EDTA blood of the patient should be sent AT ROOM TEMPERATURE in clearly labeled (with name of patient and date of birth), tightly capped tubes, wrapped in protective material, placed in a sealed plastic bag and shipped by courier service in a Styrofoam shipping box. Blood samples need to arrive in our laboratory within 48 hours after drawing. Include one blood sample as a control for shipment conditions. Please complete the test request form and send it along with the sample.
  • TAT: 3 weeks.
Dihydroxyacetone-phosphate acyltransferase (DHAPAT)

Disease

RCDP type 1, 2 and 5, and Zellweger spectrum disorders.

Description

Dihydroxyacetonephosphate-acyltransferase (DHAPAT) is the first enzyme in the etherphospholipid (plasmalogen) biosynthesis route. DHAPAT activity is deficient in rhizomelic chondrodysplasia punctata (RCDP) type 1 (PEX7 defect), type 2 (DHAPAT defect) and type 5 (PEX5L defect), as well as in Zellweger spectrum disorders.

OMIM

215100 214100 202370 222765

Method

HPLC-MS/MS

Price

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
Immunoblot peroxisomal proteins

Disease

Peroxisome biogenesis disorders.

Description

Immunoblot peroxisomal proteins is used to determine the presence and peroxisomal processing of different peroxisomal proteins. Processing of ACOX1 and peroxisomal thiolase is abnormal in Zellweger spectrum disorders, whereas in rhizomelic chondrodysplasia punctata (RCDP) type 1 only processing of peroxisomal thiolase is abnormal.

OMIM

214100 202370 215100

Method

Immunoblot

Price:

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
Immunofluorescence peroxisomal proteins

Disease

Zellweger spectrum disorders, ACOX1 deficiency, DBP deficiency and X-linked adrenoleukodystrophy (X-ALD)

Description

Immunofluorescence microscopy analysis with antibodies against peroxisomal proteins is used to determine whether in fibroblasts of the patient the morphology of peroxisomes is normal, whether there is normal import of peroxisomal matrix proteins (e.g. catalase) and whether peroxisomal membrane structures (ghosts) are present. The peroxisomal import system is disturbed in most Zellweger spectrum disorders and as a result catalase staining is generally cytosolic. Peroxisomal morphology can be also abnormal in Zellweger spectrum disorders and a subset of peroxisomal fatty acid oxidation defects (i.e. ACOX1 deficiency and DBP deficiency).

OMIM

214100 202370 264470 261515 300100

Method

Microscopy

Price

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
Peroxisomal thiolase branched-chain specific (SCPx)

Disease

SCPx deficiency.

Description

Peroxisomal thiolase branched-chain specific (SCPx) catalyzes the last step of the peroxisomal beta-oxidation of branched-chain fatty acids, including pristanic acid and C27-bile acid intermediates.

OMIM

184755

Method

HPLC-UV

Price

€ 472
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
Peroxisomal β-oxidation

Disease

Zellweger spectrum disorders , ACOX1 deficiency, DBP deficiency, AMACR deficiency, X-linked adrenoleukodystrophy (X-ALD) and SCPx deficiency.

Description

Fatty acid beta-oxidation is measured with radiolabeled C26:0 and pristanic acid, or a D3C22:0 loading test is performed. Beta-oxidation of very long-chain fatty acids (C22-C26) is deficient in Zellweger spectrum disorders, ACOX1 deficiency and DBP deficiency. Pristanic acid beta-oxidation is deficient in Zellweger spectrum disorders and DBP deficiency, and partially deficient in AMACR deficiency and SCPx deficiency.

OMIM

214100 202370 264470 261515 604489 184755

Method

Radiochemistry

Price

€ 833
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.
Phytanic acid α-oxidation

Disease

Refsum disease, rhizomelic chondrodysplasia punctata (RCDP) type 1 and Zellweger spectrum disorders.

Description

Fatty acid alpha-oxidation is measured with radiolabeled phytanic acid, and is deficient in Refsum disease (phytanoyl-CoA hydroxylase deficiency) as well as in rhizomelic chondrodysplasia punctata (RCDP) type 1 (PEX7 defect) and in Zellweger spectrum disorders.

OMIM

214100 202370 215100 266500

Method

Radiochemistry

Price

€ 833
€ 1055 for full biochemical peroxisomal screening in cultured fibroblasts.

Test request form Enzyme diagnostics

Materials:

Cultured skin fibroblasts

  • Conditions: Room temperature.
  • Specifics: Grow the fibroblasts to near confluence (~ 80% confluent) in a T-25 flask, fill the flask completely with sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) supplemented with 10% serum and 1% penicilline-streptomycine. Cap the flasks tightly and wrap the cap with parafilm. Label the flask clearly with identifying information (name of patient and date of birth), wrap the culture flaks in protective material and place it in a sealed plastic bag in the Styrofoam shipping box and ship AT ROOM TEMPERATURE by courier service. Fibroblasts need to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
    If it is not possible to culture fibroblasts from a skin biopsy in a local institute, you can send a skin biopsy in sterile buffered (with 25mM Hepes) culture medium (for example HAM or DMEM) without serum or antibiotics AT ROOM TEMPERATURE by courier service. The skin biopsy needs to arrive in our laboratory within 48 hours. Please complete the test request form and send it along with the sample.
  • TAT: 3 months.