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Improved real-time detection of the H63D and S65C mutations associated with hereditary hemochromatosis using a SimpleProbe assay format
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Corné H.W. Klaassen
Published/Copyright:
July 1, 2008
Abstract
Background: Genetic predisposition to hemochromatosis involves several different point mutations in the HFE gene. Routine testing for two such mutations (H63D and S65C) using real-time genotyping deserves special care, as these mutations are in close proximity (6 bp) of each other.
Methods: A novel assay was designed for these two mutations based on a SimpleProbe assay format that allows for more flexibility in assay design, as it requires only one detection probe instead of the two probes that are commonly used with a hybridization probe based assay.
Results: The SimpleProbe assay format yielded data that were easily interpreted without significant optimizing efforts.
Conclusions: The SimpleProbe assay format offers some unique advantages compared to hybridization probes and offers a robust and interesting alternative to hybridization probes in the detection of genetic variations. This is the first time that the use of this assay format is described in the clinical literature.
Clin Chem Lab Med 2008;46:985–6.
Received: 2008-1-28
Accepted: 2008-3-26
Published Online: 2008-07-01
Published in Print: 2008-07-01
©2008 by Walter de Gruyter Berlin New York
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Keywords for this article
hemochromatosis;
hybridization probes;
LightCycler;
mutation detection;
SimpleProbe
Articles in the same Issue
- Editorial
- CCLM: Evolving to meet the needs of today's laboratory professionals and scientists
- Bacterial detection and blood product contamination
- Editorial: Rapid diagnostic tests to detect pathogenic microorganisms
- Reviews
- Rapid methods for diagnosis of bloodstream infections
- Validation criteria for nucleic acid amplification techniques for bacterial infections
- Incidence of bacterial transmission and transfusion reactions by blood components
- Strategies of bacteria screening in cellular blood components
- Methods for the detection of bacterial contamination in blood products
- Bacteria detection by flow cytometry
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- Screening for genetic heterogeneity in the interferon sensitivity determining region of the hepatitis C virus genome by polymerase chain reaction with melting curve analysis
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