HFE mpx RealFast™ Assay: A Precise Tool for Hereditary Haemochromatosis Detection

Introduction

Hereditary haemochromatosis (HH) is a prevalent genetic disorder, particularly among individuals of Northern European descent. This condition is characterized by the excessive absorption and accumulation of iron in various organs. HH can lead to severe health issues, including liver cirrhosis, liver cancer, diabetes, arthritis, and cardiopathies. Early diagnosis and treatment are crucial to prevent irreversible organ damage. ViennaLab Diagnostics GmbH has developed the HFE mpx RealFast™ Assay, a powerful tool for detecting two common mutations associated with HH: H63D and C282Y in the HFE gene.

Understanding Hereditary Haemochromatosis

HH encompasses a group of genetic disorders with diverse molecular defects and clinical manifestations. These disorders share a common feature: a relative deficiency of hepcidin, a hormone responsible for regulating systemic iron levels. Mutations in genes like HFE, Transferrin receptor 2, Hemojuvelin, Hepcidin, and Ferroportin 1 disrupt hepcidin regulation, contributing to iron overload. Among these, mutations in the HFE gene account for the most common form of HH.

The HFE gene has two prevalent mutations associated with HH: H63D and C282Y. Approximately 80% of HH patients are homozygous for the C282Y mutation, while fewer individuals are compound heterozygous for C282Y and H63D mutations. Homozygous carriers of H63D mutations typically have minimal increases in iron absorption and rarely develop HH.

The Role of Genetic Testing

Genetic testing for HFE mutations plays a pivotal role in HH diagnosis. Identifying individuals carrying these mutations allows for early intervention and prevention of iron-related complications. The HFE mpx RealFast™ Assay offers a fast and precise method for detecting H63D and C282Y mutations in the HFE gene.

Storage and Stability

The kit is shipped on cooling blocks and should be stored at -30 to -15°C upon receipt. Alternatively, it can be stored at 2 to 8°C for short-term use within one month. The HFE mpx RealFast™ Assay kit can withstand up to 20 freeze/thaw cycles without losing its activity. Avoid prolonged exposure to intense light to maintain its stability.

Principle of the Test

The HFE mpx RealFast™ Assay relies on the fluorogenic 5’ nuclease assay, commonly known as the TaqMan® assay. Each reaction contains two gene-specific primer pairs, which amplify fragments of the HFE gene, as well as four dual-labeled, allele-specific hydrolysis probes that bind to the target sequences of the amplified fragments. During the extension phase of PCR, the Taq DNA polymerase cleaves the 5’-fluorescent reporter from the hybridized probe. This separation generates a fluorescent signal in real-time, proportional to the PCR product.

Conclusion

Hereditary haemochromatosis is a prevalent genetic disorder in Northern European populations, and early diagnosis is critical for preventing severe health complications. ViennaLab’s HFE mpx RealFast™ Assay offers a fast, precise, and multiplexed method for detecting H63D and C282Y mutations in the HFE gene. With its ability to discriminate between normal and mutant genotypes, this assay empowers healthcare professionals to identify HH patients promptly and initiate life-saving interventions. Early diagnosis and intervention can significantly improve the quality of life for individuals affected by HH.