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Apolipoprotein E genotyping: a comparative study between restriction endonuclease mapping and allelic discrimination with the LightCycler.

Sabrina Ballerini·Lorenza Bellincampi·Sergio Bernardini·Stefania Casciani·Corradino Motti·Claudio Cortese·Giorgio Federici
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Abstract

BACKGROUND: The apolipoprotein E (apo E) polymorphism is associated with the risk of developing cardiovascular disease and the risk and the time of onset of Alzheimer's disease. Therefore, the interest in apo E genotyping is high, both for epidemiological research and for the purpose of diagnosing dyslipidemia or dementia. The aim of our study was to compare and evaluate two different methods for apo E genotyping, both on the basis of polymerase chain reaction (PCR). METHODS: Genomic DNA of 197 subjects was extracted from whole blood. The first method involved DNA amplification performing a PCR using specific primers and endonuclease restriction mapping. The second one was a DNA assay that used real-time PCR on the LightCycler instrument (Roche). RESULTS: We obtained a 100% concordance between the two methods and we found a relative allelic frequency distribution typical for an Italian population. CONCLUSIONS: The LightCycler (LC) allelic discrimination method for apo E genotyping seems to be rapid, simple and accurate, suggesting a possible successful use of this method for diagnostic purposes.

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AllelesApolipoproteins EDeoxyribonucleases, Type II Site-SpecificHumansPolymerase Chain ReactionRestriction MappingComparative StudyJournal ArticleResearch Support, Non-U.S. Gov't

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Sabrina Ballerini, Lorenza Bellincampi, Sergio Bernardini, Stefania Casciani, Corradino Motti, Claudio Cortese, Giorgio Federici. 2002. Apolipoprotein E genotyping: a comparative study between restriction endonuclease mapping and allelic discrimination with the LightCycler.. https://doi.org/10.1016/s0009-8981(01)00769-0

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