Validation of the Hemochromatosis (2SNP+) Direct EUROArray Assay for the Molecular Diagnosis of -Related Hereditary Hemochromatosis.

Two missense variants in the gene, c.845G>A (p.Cys282Tyr) and c.187C>G (p.His63Asp), are commonly screened as part of the diagnostic workup for -related hereditary hemochromatosis (HH) and iron overload. Identification of the two variants can be achieved by polymerase chain reaction (PCR)-based laboratory tests and other methods. Evaluation of the analytical performance of the test is essential to ensure that the assay is precise and accurate. The aim of this study was to evaluate the analytical performance of the DNA microarray-based Hemochromatosis (2SNP+) Direct assay on the EUROArray test system (EUROIMMUN, Lübeck, Germany). Evaluation of the commercial assay was performed on 50 clinical blood samples and 26 retrospective College of American Pathologists (CAP)-provided external quality assurance (EQA) DNA samples and compared to a laboratory-developed PCR-restriction enzyme digestion (PCR-RE) test and DNA sequencing. genotyping results obtained from both Hemochromatosis (2SNP+) Direct and PCR-RE assays were 100% concordant with nucleotide sequencing for all clinical samples evaluated. One hundred percent accuracy was also achieved on the retrospective CAP EQA samples. Precision studies performed on wild type and c.845G>A/c.187C>G compound heterozygous whole blood samples showed 100% intra-run repeatability ( = 3) and 100% inter-run reproducibility ( = 3), respectively. The Hemochromatosis (2SNP+) Direct EUROArray test provides a rapid and accurate method of detection for both the c.845G>A and c.187C>G variants for molecular diagnosis of -related HH.