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26817 Hereditary Hemochromatosis, HFE Variant Analysis (LBOR0250)

Hereditary Hemochromatosis, HFE Variant Analysis (LBOR0250)
Test Code: LBOR0250
Synonyms/Keywords

Hereditary Hemochromatosis, HFE, C282Y, H63D, serum transferrin-iron saturation, serum ferritin, iron overload

Test Components

​​Variants assessed in HFE (NM_000410.4): c.845G>A (p.C282Y), c.187C>G (p.H63D), and c.193A>T (p.S65C) 

Useful For

To establish or confirm a clinical diagnosis of hereditary hemochromatosis (HH) in individuals with increased serum transferrin-iron saturation and serum ferritin studies, To screen adult family members of individuals with HH due to HFE C282Y and/or H63D, NOT useful for population or carrier screening of HFE variants.

HFE-related Hereditary Hemochromatosis (HH) is an autosomal recessive disorder characterized by increased iron storage. Up to 90% of HFE-related HH in Northern Europeans is caused by homozygosity for C282Y and the next most common cause is due to compound heterozygosity for C282Y/H63D. HFE-related HH follows reduced penetrance. Individuals may experience no symptoms, biochemical symptoms (elevated serum transferrin-iron saturation and serum ferritin studies), and or clinical symptoms. Initial symptoms may include fatigue, joint and abdominal pain, skin hyperpigmentation, and weight loss. Further symptoms may develop due to iron accumulation including arthritis, liver cirrhosis or liver cancer, diabetes, or heart disease.  

Specimen Requirements
Fasting Required
Specimen TypePreferred Container/TubeAcceptable Container/TubeSpecimen VolumeSpecimen Minimum Volume
(allows for 1 repeat)
Pediatric Minimum Volume
(no repeat)
​No
​Whole Blood
​Lavender EDTA tube
​4 mL
​1.0 mL​
Collection Processing Instructions

Send in original tube. Do not aliquot or share for other testing.

Clients will be contacted to provide additional specimen if DNA quality is insufficient. An alternative specimen type may also be requested, at the discretion of the laboratory.

Specimen Stability Information
Specimen Type
TemperatureTime
​​Whole Blood




​Room Temp
​72 hours
​Refrigerated
(Preferred)
​14 days
​Frozen
​Not Acceptable
Interference

This test is contraindicated for patients who have undergone an allogenic stem cell transplant (e.g., bone marrow or peripheral stem cell from a donor). 

* All specimens will be evaluated at the Sanford Medical Genetics laboratory for test suitability. 

Limitations:Rare variants in the HFE gene and other genetic factors associated with HH are not assessed by this assay. Additionally, rare genetic polymorphisms may interfere with variant assessment and lead to inconclusive results. Test results should be interpreted in the context of clinical findings, other laboratory data, family history, and environmental and lifestyle risk factors. If clinically warranted, additional testing should be considered. Allogenic bone marrow transplants may result in false-positive or false-negative results.    

Test algorithm guidelines have been established and a Sanford customized algorithm is below (Kowdley et al, 2019).

https://sanfordlaboratories.testcatalog.org/catalogs/455/files/17270
Variants are assessed with quantitative Polymerase Chain Reaction (qPCR) using TaqMan probes. Sanger sequencing may be used to confirm variants.
Note: S65C is only reported when C282Y is identified in a heterozygous state due to its limited biochemical relevance. Studies have demonstrated that although some compound C282Y/S65C heterozygous individuals may present with increased serum iron and ferritin levels, they do not present with clinical manifestations of hemochromatosis nor iron-associated liver fibrosis (Holstrom, 2002; Pederson, 2009). Moreover, in 2019 American College of Gastroenterology (ACG) classified S65C as a polymorphism without clinical significance (Kowdley, 2019).

Performing Laboratory Information
Performing Location
Day(s) Test PerformedAnalytical TimeMethodology/Instrumentation
​Sanford Laboratories
Sioux Falls
​Monday-Friday
​8 days
Quantitative Polymerase Chain Reaction (qPCR) using TaqMan probes
Outreach CPTs
CPTModifier
(if needed)
QuantityDescriptionComments
Synonyms/Keywords

Hereditary Hemochromatosis, HFE, C282Y, H63D, serum transferrin-iron saturation, serum ferritin, iron overload

Test Components

​​Variants assessed in HFE (NM_000410.4): c.845G>A (p.C282Y), c.187C>G (p.H63D), and c.193A>T (p.S65C) 

Ordering Applications
Ordering ApplicationDescription
If the ordering application you are looking for is not listed, contact your local laboratory for assistance.
Specimen Requirements
Fasting Required
Specimen TypePreferred Container/TubeAcceptable Container/TubeSpecimen VolumeSpecimen Minimum Volume
(allows for 1 repeat)
Pediatric Minimum Volume
(no repeat)
​No
​Whole Blood
​Lavender EDTA tube
​4 mL
​1.0 mL​
Collection Processing

Send in original tube. Do not aliquot or share for other testing.

Clients will be contacted to provide additional specimen if DNA quality is insufficient. An alternative specimen type may also be requested, at the discretion of the laboratory.

Specimen Stability Information
Specimen Type
TemperatureTime
​​Whole Blood




​Room Temp
​72 hours
​Refrigerated
(Preferred)
​14 days
​Frozen
​Not Acceptable
Interference

This test is contraindicated for patients who have undergone an allogenic stem cell transplant (e.g., bone marrow or peripheral stem cell from a donor). 

* All specimens will be evaluated at the Sanford Medical Genetics laboratory for test suitability. 

Limitations:Rare variants in the HFE gene and other genetic factors associated with HH are not assessed by this assay. Additionally, rare genetic polymorphisms may interfere with variant assessment and lead to inconclusive results. Test results should be interpreted in the context of clinical findings, other laboratory data, family history, and environmental and lifestyle risk factors. If clinically warranted, additional testing should be considered. Allogenic bone marrow transplants may result in false-positive or false-negative results.    

Test algorithm guidelines have been established and a Sanford customized algorithm is below (Kowdley et al, 2019).

https://sanfordlaboratories.testcatalog.org/catalogs/455/files/17270
Variants are assessed with quantitative Polymerase Chain Reaction (qPCR) using TaqMan probes. Sanger sequencing may be used to confirm variants.
Note: S65C is only reported when C282Y is identified in a heterozygous state due to its limited biochemical relevance. Studies have demonstrated that although some compound C282Y/S65C heterozygous individuals may present with increased serum iron and ferritin levels, they do not present with clinical manifestations of hemochromatosis nor iron-associated liver fibrosis (Holstrom, 2002; Pederson, 2009). Moreover, in 2019 American College of Gastroenterology (ACG) classified S65C as a polymorphism without clinical significance (Kowdley, 2019).

Useful For

To establish or confirm a clinical diagnosis of hereditary hemochromatosis (HH) in individuals with increased serum transferrin-iron saturation and serum ferritin studies, To screen adult family members of individuals with HH due to HFE C282Y and/or H63D, NOT useful for population or carrier screening of HFE variants.

HFE-related Hereditary Hemochromatosis (HH) is an autosomal recessive disorder characterized by increased iron storage. Up to 90% of HFE-related HH in Northern Europeans is caused by homozygosity for C282Y and the next most common cause is due to compound heterozygosity for C282Y/H63D. HFE-related HH follows reduced penetrance. Individuals may experience no symptoms, biochemical symptoms (elevated serum transferrin-iron saturation and serum ferritin studies), and or clinical symptoms. Initial symptoms may include fatigue, joint and abdominal pain, skin hyperpigmentation, and weight loss. Further symptoms may develop due to iron accumulation including arthritis, liver cirrhosis or liver cancer, diabetes, or heart disease.  

Test Components

​​Variants assessed in HFE (NM_000410.4): c.845G>A (p.C282Y), c.187C>G (p.H63D), and c.193A>T (p.S65C) 

For more information visit:
Performing Laboratory Information
Performing Location
Day(s) Test PerformedAnalytical TimeMethodology/Instrumentation
​Sanford Laboratories
Sioux Falls
​Monday-Friday
​8 days
Quantitative Polymerase Chain Reaction (qPCR) using TaqMan probes
For billing questions, see Contacts
Outreach CPTs
CPTModifier
(if needed)
QuantityDescriptionComments
For most current information refer to the Marshfield Laboratory online reference manual.