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Test ID LBOR0250 Hereditary Hemochromatosis, HFE Variant Analysis

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  

AKA

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

Specimen Type/Requirements

Whole blood in lavender top (EDTA) tube  

Send in original tube, do not aliquot and do not share for other testing. 

  

  • 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). 

  • 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.  

  • All specimens should be sent in the original container and should not be aliquoted to another tube.  

  • In addition, the specimen submitted should ONLY be used for this testing and should not be shared with any other testing that would also utilize this specimen type. 

Specimen Volume

 Preferred Volume     2 - 4 mL   
 Minimum Vollume   1 mL   

 

Stability/Transport

 Room Temperature     72 hours       
 Refrigerated   14 days     Preferred for transport   
 Frozen  Not Acceptable     

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

Performed Test Frequency

Monday through Friday 

Report Available

7 days

Additional Information

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.  

 

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.    

References 

  1. Bacon BR, Adams PC, Kowdley KV, Powell LW, Tavill AS; American Association for the Study of Liver Diseases. Diagnosis and management of hemochromatosis: 2011 practice guideline by the American Association for the Study of Liver Diseases. Hepatology. 2011 Jul;54(1):328-43. doi: 10.1002/hep.24330. PMID: 21452290; PMCID: PMC3149125. 

  1. Holmström P, Marmur J, Eggertsen G, Gåfvels M, Stål P. Mild iron overload in patients carrying the HFE S65C gene mutation: a retrospective study in patients with suspected iron overload and healthy controls. Gut. 2002 Nov;51(5):723-30. doi: 10.1136/gut.51.5.723. PMID: 12377814; PMCID: PMC1773427. 

  1. Kowdley KV, Brown KE, Ahn J, Sundaram V. ACG Clinical Guideline: Hereditary Hemochromatosis. Am J Gastroenterol. 2019 Aug;114(8):1202-1218. doi: 10.14309/ajg.0000000000000315. Erratum in: Am J Gastroenterol. 2019 Dec;114(12):1927. doi: 10.14309/ajg.0000000000000469. PMID: 31335359. 

  1. Pedersen P, Milman N. Extrinsic factors modifying expressivity of the HFE variant C282Y, H63D, S65C phenotypes in 1,294 Danish men. Ann Hematol. 2009 Oct;88(10):957-65. doi: 10.1007/s00277-009-0714-x. Epub 2009 Mar 7. PMID: 19271219. 

  1. Porto G, Brissot P, Swinkels DW, Zoller H, Kamarainen O, Patton S, Alonso I, Morris M, Keeney S. EMQN best practice guidelines for the molecular genetic diagnosis of hereditary hemochromatosis (HH). Eur J Hum Genet. 2016 Apr;24(4):479-95. doi: 10.1038/ejhg.2015.128. Epub 2015 Jul 8. PMID: 26153218; PMCID: PMC4929861. 

Methodology

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

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 Lab

Sanford Medical Genetics Laboratory - Sioux Falls

CPT

81256

Coverage Limitations

For non-Medicare patients, a separate completed and signed waiver is required to accompany the specimen: 
Commercial Insurance Patient Waiver of Liability (Non-Medicare) Molecular Genetics

Testing Algorithm

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

Hereditary Hemochromatosis Algorithm