Clinical evaluation of a hemochromatosis next‐generation sequencing gene panel

Lanktree, Matthew B.; Sadiković, Bekim; Waye, John S.; Levstik, Alexander; Lanktree, Bruce B.; Yudin, Jovana; Crowther, Mark; Paré, Guillaume; Adams, Paul C.

doi:10.1111/ejh.12820

Cited by 21 publications

(18 citation statements)

References 25 publications

(28 reference statements)

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“…This is partly due to environmental factors, including physiological blood loss, as well as the interaction of other genetic factors . Novel research is exploiting next‐generation sequencing techniques to identify these disease modifying‐genes that may close the gap in knowledge about incomplete penetrance of the C282Y substitution. At present, a definitive test to determine whether a person with the homozygous C282Y substitution will develop clinical symptoms remains elusive.…”

Section: Introductionmentioning

confidence: 99%

Haemochromatosis: a clinical update for the practising physician

Radford-Smith

Powell

2018

Internal Medicine Journal

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Haemochromatosis is most commonly due to the autosomal recessive inheritance of a C282Y substitution in the HFE protein, whereby both alleles of the corresponding gene are affected. The disease is characterised by an inappropriate increase in intestinal iron absorption due to reduced expression of the iron regulatory protein, hepcidin. Progressive iron deposition in parenchymal tissues may ultimately lead to liver and other organ toxicity. The characteristic biochemical abnormalities are raised serum ferritin and transferrin saturation, which can be used in conjunction with genetic tests and emerging magnetic resonance imaging-based techniques to diagnose patients with the disorder. Progressive iron overload can manifest clinically as advanced fibrosis, cirrhosis and hepatocellular carcinoma. Enigmatically, the penetrance of both raised iron indices and clinically significant disease is incomplete in patients with hereditary haemochromatosis. Regardless, advanced clinical presentations of the disease have become less common due to increased awareness and earlier diagnosis. On the other hand, obesity and alcohol have been identified as major risk factors that can compound the risk of liver injury in people with hereditary (HFE) haemochromatosis. The prospect of modifying genes that may contribute to the clinical expression of the disease is the subject of ongoing research. Treatment with phlebotomy remains the first-line therapy, and if instigated early leads to a normal life expectancy. A healthy, well-balanced diet is recommended to be incorporated as part of the ongoing management of the disease.

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Section: Introductionmentioning

confidence: 99%

Haemochromatosis: a clinical update for the practising physician

Radford-Smith

Powell

2018

Internal Medicine Journal

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“…This report shows that the multiplex assay outperforms the singleplex assays in terms of throughput and costs, while maintaining highly accurate and easily interpretable data. Our assay is also highly competitive compared to other published approaches that are used for genotyping of H63D and/or C282Y mutations in HFE (15,(23)(24)(25)(26)(27)(28)(29). Examples of alternative screening methods to detect H63D and C282Y mutations are next-generation sequencing with a targeted panel for hemochromatosis (28), real-time PCR (15,29), Sanger sequencing (15), multiplex linked linear amplification coupled with allele-specific oligonucleotide hybridization (27), restriction fragment length polymorphism analysis (15,26,29), single base extension assays, and other specific amplification methods that are combined with capillary electrophoresis (13), reverse dot-blot analysis (25), heteroduplex technology (24), and single-strand conformation polymorphism analysis (23).…”

Section: Discussionmentioning

confidence: 98%

“…A limitation of our method is that it is limited to the analysis of the recurrent H63D and C282Y HFE mutations while clinical mutations in HFE and other genes have also been reported in patients with iron overload, albeit less frequently (28,30). Another limitation of our assay is that it is based on PCR, which is susceptible to allele dropout caused by polymorphisms; however, we minimized the chances of assay interference by incorporating mixed bases in locations of known SNPs (rs570212174 and rs376650371) in the primers (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Multiplex Allele-Specific PCR for Simultaneous Detection of H63D and C282Y HFE Mutations in Hereditary Hemochromatosis

Arts

Waye

2018

The Journal of Applied Laboratory Medicine

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Background Hereditary hemochromatosis (HH) is characterized by excessive iron absorption in the intestine, which can lead to failure of vital organs such as the heart, liver, and pancreas. Among northern Europeans, HH is most often associated with the C282Y and H63D mutations of the HFE gene. We developed a test that allows screening for both mutations in a single reaction. Methods A multiplex allele-specific PCR was developed for simultaneous genotyping of the H63D and C282Y HFE mutations. PCR fragments were designed such that the resulting PCR product can be analyzed in a single polyacrylamide gel lane. Results Test results from our multiplex assay were concordant with genotypes of 55 Canadian patients with suspected hemochromatosis, which had previously been established by allele-specific PCRs that targeted H63D and C282Y in separate reactions. Conclusions Molecular diagnostic detection of H63D and C282Y mutations can be achieved by a variety of methods, but these are not necessarily time-efficient or economical. Multiplex allele-specific PCR is an excellent tool for molecular diagnostic screening for H63D and C282Y mutations in patients with suspected hemochromatosis. This method is inexpensive, accurate, and highly efficient in terms of labor, throughput, and turnaround time.

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“…Next-generation sequencing may provide further tools for those with more uncommon mutations predisposing to iron overload. 10 Whole genome sequencing can provide more detailed data to associate mutations with iron overload. 11 Secondary iron overload is related to frequent blood transfusions, exogenous iron intake, or certain hematological diseases such as sickle cell anemia or thalassemia.…”

Section: Genetics and Risk Factorsmentioning

confidence: 99%

Diagnosis and Management of Genetic Iron Overload Disorders

et al. 2018

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Iron overload disorders lead to excess iron deposition in the body, which can occur as a result of genetic or secondary causes. Genetic iron overload, referred to as hereditary hemochromatosis, may present as a common autosomal recessive mutation or as one of several uncommon mutations. Secondary iron overload may result from frequent blood transfusions, exogenous iron intake, or certain hematological diseases such as dyserythropoietic syndrome or chronic hemolytic anemia. Iron overload may be asymptomatic, or may present with significant diseases of the liver, heart, endocrine glands, joints, or other organs. If treated appropriately prior to end-organ damage, life expectancy has been shown to be similar compared to matched populations. Alongside clinical assessment, diagnostic studies involve blood tests, imaging, and in some cases liver biopsy. The mainstay of therapy is periodic phlebotomy, although oral chelation is an option for selected patients.

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Clinical evaluation of a hemochromatosis next‐generation sequencing gene panel

Cited by 21 publications

References 25 publications

Haemochromatosis: a clinical update for the practising physician

Haemochromatosis: a clinical update for the practising physician

Multiplex Allele-Specific PCR for Simultaneous Detection of H63D and C282Y HFE Mutations in Hereditary Hemochromatosis

Diagnosis and Management of Genetic Iron Overload Disorders

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