Apolipoprotein E genotyping: a comparative study between restriction endonuclease mapping and allelic discrimination with the LightCycler.
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.
Ballerini
Sabrina
S
Department of Internal Medicine, University of Rome Tor Vergata, Rome, Italy.
Bellincampi
Lorenza
L
Bernardini
Sergio
S
Casciani
Stefania
S
Motti
Corradino
C
Cortese
Claudio
C
Federici
Giorgio
G
D000483:Alleles; D001057:Apolipoproteins E / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping / Q000379:methods*
D001057:Apolipoproteins E; D015252:Deoxyribonucleases, Type II Site-Specific; C037130:GCGC-specific type II deoxyribonucleases