J Med Life Sci > Volume 12(1); 2015 > Article
Journal of Medicine and Life Science 2015;12(1):30-34.
DOI: https://doi.org/10.22730/jmls.2015.12.1.30    Published online December 18, 2018.
Real-Time Quantitative Polymerase Chain Reaction for Apolipoprotein E genotyping
Young Hwan Park, Young Ree Kim, Sun Hyung Kim, Sung Ha Kang
Correspondence:  Young Ree Kim,
Abstract
Background : Apolipoprotein E (ApoE) is an important substance responsible for the lipoprotein metabolism and the transportation of lipid and known to play an important role in the body's physiological and pathological processes. Recently, ApoE genotyping test has been applied to the field of diagnostic methods for predicting high risk of Alzheimer's disease, cardiovascular diseases, as well as new diseases such as immune regulation disorders. Therefore, a faster and more accurate ApoE genotyping test is required for the prevention of related disease and the general public health. Methods : We evaluated 220 serum specimens using two kinds of assays: one-stop Real-Q ApoE genotyping (BioSewoom, Seoul, Korea) and Seeplex ApoE ACE genotyping (Seegene, Seoul, Korea). The results were finally confirmed using sequencing as the reference. Statistical analysis was performed for the concordance rates between each assay and sequencing and the sensitivity and specificity were calculated with their 95% confidence intervals. Results : Sequencing results of 220 DNA samples showed 1 case of E2/E2, 139 of E3/E3, 9 of E4/E4, 17 of E2/E3, 4 of E2/E4, 50 of E3/E4. The concordance rate between One-step Real-Q ApoE genotyping and sequencing was 100% (220/220), and that between Seeplex ApoE ACE genotyping and sequencing was 99.5% (219/220). Both sensitivity and specificity of Onestep Real-Q ApoE genotyping for every genotypes was 100%. The sensitivities and specificities of Seeplex ApoE ACE genotyping were almost 100%, with two exception: 99.3% sensitivity in E3/E3 and 99.5% specificity in E2/E2. Conclusions : The performances of One-step Real-Q ApoE genotyping and Seeplex ApoE ACE genotyping were comparable overall. However, the former showed better sensitivity and specificity comparable with those of sequencing. Hence, One-step ApoE genotyping could be a faster and more accurate option for ApoE genotyping.
Key Words: One-step ApoE Genotyping, Sequencing, Concordance rate
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