Apolipoprotein E genotyping in patients with neurodegenerative diseases.
OBJECTIVE
There is a growing demand to perform apolipoprotein E (Apo-E) genotyping on neuropathologic archive material. Due to the extremely long fixation time, this material is unsuitable for routinely used Apo-E genotyping methods. We present an investigation into the applicability of a new method.


METHODS
An Apo-E genotyping method was tested for use on formalin-fixed paraffin-embedded brain tissue, using semi-nested PCR followed by hybridization with biotin-labeled allele-specific oligonucleotides, and chemiluminescent detection. The method was applied to 88 archive samples of different neurologic disorders.


RESULTS
With this technique 76% (67/88) of the samples could be genotyped. The crucial step is the semi-nested PCR. All the samples from which a PCR product could be obtained, the Apo-E gene could be genotyped without interpretation problems. Seventy-six percent of the samples that could not be genotyped, were fixed in unbuffered formalin.


CONCLUSIONS
This technique offers a good Apo-E genotyping method applicable on neuropathological archive material in order to support in retrospect clinical studies.
Ballering
L A
LA
Department of Clinical Chemistry, Medical Spectrum Twente, Hospital Group, Enschede, The Netherlands.
Steffens-Nakken
H M
HM
Esselink
R A
RA
De Vos
R A
RA
Steur
E N
EN
Vermes
I
I
D000544:Alzheimer Disease / Q000235:genetics* / Q000378:metabolism; D001057:Apolipoproteins E / Q000235:genetics*; D005838:Genotype; D006801:Humans; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods
D001057:Apolipoproteins E