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Haemochromatosis – H63D, S65C, C282Y mutation in the HFE gene

Test covered by the reimbursement:

YES

Clinical expertise code:

208

Test without reimbursement:

YES

Gender:

Woman/Man

Material:

Peripheral blood, Buccal swab …

Turnover time:

3 weeks

STATIM:

3 days

Material:

Peripheral blood | 1x 3 ml of whole blood in K3 EDTA tube

Storage after examination: week after the report is issued 2 – 8°C

Buccal swab | 2x swab stick for buccal swab collection

Storage after examination: week after the report is issued 2 – 8°C

Isolated DNA from blood | 10–100 ng/μL of isolated DNA from blood in a PCR tube of at least 15 μL.

Storage after examination: stored in a DNA archive without restriction 15°C

Chorionic villi | Chorionic villi, min. 30 mg tissue in a microtube (Eppendorf type)

Storage after examination: week after the report is issued 2 – 8°C

Amniotic fluid | 3x 10 ml of amniotic fluid in a tube

Storage after examination: week after the report is issued 2 – 8°C

Cord blood | 2–3 ml of cord blood in EDTA

Storage after examination: week after the report is issued 2 – 8°C

Conception product | Foetal tissue in saline

Storage after examination: 1 aliquot is stored -25°C

Cultured cells | 1.5 ml of cultured cells in a microtube (Eppendorf type)

Storage after examination: 180 days 2 – 8°C

Isolated DNA from cordocentesis | 30–100 ng/μL of isolated DNA from cordocentesis in a microtube (Eppendorf type)

Storage after examination: stored in a DNA archive without restriction 15°C

Isolated DNA from chorionic villi | 30–100 ng/μL of isolated DNA from chorionic villi in a microtube (Eppendorf type)

Storage after examination: stored in a DNA archive without restriction 15°C

DNA isolated from the product of conception | 50–100 ng/μL in microtube (Eppendorf type)

Storage after examination: stored in a DNA archive without restriction 15°C

Isolated DNA from amniotic fluid | 30–100 ng/μL of isolated DNA from amniocentesis in a microtube (Eppendorf type)

Storage after examination: stored in a DNA archive without restriction 15°C

Quick test description:

HFE gene testing by real-time PCR is performed to detect the most common p.C282Y, p.H63D and p.S65C mutations associated with hereditary haemochromatosis.

Test details:

The HFE gene encodes a protein that binds to the transferrin receptor (TFR), reducing its affinity (binding strength) for iron-bearing transfers and therefore regulating iron accumulation in tissue. In HFE deficiency, iron is highly absorbed and accumulated for a long time in the liver, skin, pancreas, endocrine glands, joints and heart with manifestations of the hereditary haemochromatosis. Symptoms of haemochromatosis (fatigue, nausea, joint pain, decreased libido and fertility) can occur between 30–50 years of age, more often in men.

Iron accumulation in the liver can lead to cirrhosis and hepatocellular carcinoma. Iron accumulation in the pancreas can manifest as type II diabetes with typical skin colouration (bronze diabetes). Heartbeat may be irregular (arrhythmia). Iron can also accumulate in other organs (nervous system, adrenal glands, pituitary gland, ear). Those affected by haemochromatosis are less likely to tolerate some infectious diseases. Mutations in other modifying genes together with external environmental influences may be involved in the development of the disease. Only 23–25% of carriers of significant HFE gene mutations therefore have symptoms of haemochromatosis. Women of childbearing potential are more protected from iron accumulation by regular menstrual blood loss, and symptoms of haemochromatosis can occur only during menopause.

The most common p.C282Y mutation in heterozygous form is carried by 8–10% of the population of Western, Northern and Central Europe. The other 2 mutations, p.H63D and p.S65C, occur less frequently. HFE gene testing is performed to detect the most common mutations (p.C282Y, p.H63D and p.S65C) using real-time PCR. This method enables monitoring of the progress of the amplification reaction at any point. Each individual cycle has the same properties as classical PCR, i.e. denaturation of double-stranded DNA, primer mounting, chain extension by Taq DNA polymerase. When annealing, not only the appropriate primers are used, but also probes (specific DNA oligonucleotide sequences) with fluorophores. Detection selectivity is ensured by specific probes.

Indication

clinical suspicion of haemochromatosis (increased serum iron concentration, increased serum ferritin concentration, hepatomegaly, diabetes mellitus, skin hyperpigmentation), evidence of haemochromatosis or HFE gene mutation in a relative